US20110150902A1 - Treating cancer - Google Patents
Treating cancer Download PDFInfo
- Publication number
- US20110150902A1 US20110150902A1 US12/979,105 US97910510A US2011150902A1 US 20110150902 A1 US20110150902 A1 US 20110150902A1 US 97910510 A US97910510 A US 97910510A US 2011150902 A1 US2011150902 A1 US 2011150902A1
- Authority
- US
- United States
- Prior art keywords
- cancer
- melanoma
- patients
- cells
- chronic inflammation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 166
- 201000011510 cancer Diseases 0.000 title claims abstract description 109
- 201000001441 melanoma Diseases 0.000 claims abstract description 85
- 238000011282 treatment Methods 0.000 claims abstract description 83
- 208000037976 chronic inflammation Diseases 0.000 claims abstract description 59
- 238000000034 method Methods 0.000 claims abstract description 44
- 229940022399 cancer vaccine Drugs 0.000 claims abstract description 25
- 238000009566 cancer vaccine Methods 0.000 claims abstract description 24
- 238000002512 chemotherapy Methods 0.000 claims abstract description 14
- 230000006020 chronic inflammation Effects 0.000 claims description 43
- 241000124008 Mammalia Species 0.000 claims description 37
- 101000578784 Homo sapiens Melanoma antigen recognized by T-cells 1 Proteins 0.000 claims description 14
- 102100028389 Melanoma antigen recognized by T-cells 1 Human genes 0.000 claims description 14
- 102100022430 Melanocyte protein PMEL Human genes 0.000 claims description 12
- 101800001271 Surface protein Proteins 0.000 claims description 12
- 108010002687 Survivin Proteins 0.000 claims description 5
- 230000005855 radiation Effects 0.000 claims description 4
- 150000003431 steroids Chemical class 0.000 claims description 4
- 102000000763 Survivin Human genes 0.000 claims 2
- 239000000463 material Substances 0.000 abstract description 16
- 102000004127 Cytokines Human genes 0.000 description 58
- 108090000695 Cytokines Proteins 0.000 description 58
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 description 46
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 46
- 210000004027 cell Anatomy 0.000 description 44
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 44
- 239000003795 chemical substances by application Substances 0.000 description 37
- 108090000978 Interleukin-4 Proteins 0.000 description 30
- 102000004388 Interleukin-4 Human genes 0.000 description 30
- 206010027480 Metastatic malignant melanoma Diseases 0.000 description 30
- 208000021039 metastatic melanoma Diseases 0.000 description 30
- 210000001151 cytotoxic T lymphocyte Anatomy 0.000 description 25
- 101000716102 Homo sapiens T-cell surface glycoprotein CD4 Proteins 0.000 description 23
- 102100036011 T-cell surface glycoprotein CD4 Human genes 0.000 description 23
- 230000009885 systemic effect Effects 0.000 description 22
- 238000004458 analytical method Methods 0.000 description 21
- 239000000427 antigen Substances 0.000 description 19
- 102000036639 antigens Human genes 0.000 description 19
- 108091007433 antigens Proteins 0.000 description 19
- 230000014509 gene expression Effects 0.000 description 19
- 101000808011 Homo sapiens Vascular endothelial growth factor A Proteins 0.000 description 17
- 108090000176 Interleukin-13 Proteins 0.000 description 17
- 102000003816 Interleukin-13 Human genes 0.000 description 17
- 210000001744 T-lymphocyte Anatomy 0.000 description 17
- 208000026278 immune system disease Diseases 0.000 description 17
- 230000001404 mediated effect Effects 0.000 description 17
- 210000001519 tissue Anatomy 0.000 description 17
- 102000017420 CD3 protein, epsilon/gamma/delta subunit Human genes 0.000 description 15
- 108050005493 CD3 protein, epsilon/gamma/delta subunit Proteins 0.000 description 15
- 238000003556 assay Methods 0.000 description 15
- 210000004369 blood Anatomy 0.000 description 15
- 239000008280 blood Substances 0.000 description 15
- 230000000694 effects Effects 0.000 description 15
- 238000000684 flow cytometry Methods 0.000 description 15
- 238000000338 in vitro Methods 0.000 description 15
- 102100039037 Vascular endothelial growth factor A Human genes 0.000 description 14
- 102100037850 Interferon gamma Human genes 0.000 description 13
- 108010074328 Interferon-gamma Proteins 0.000 description 13
- 238000002560 therapeutic procedure Methods 0.000 description 13
- -1 IL-1β Proteins 0.000 description 12
- 208000007256 Nevus Diseases 0.000 description 12
- 239000011324 bead Substances 0.000 description 12
- 238000002474 experimental method Methods 0.000 description 12
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 description 12
- 208000000453 Skin Neoplasms Diseases 0.000 description 11
- 190000008236 carboplatin Chemical compound 0.000 description 11
- 229960004562 carboplatin Drugs 0.000 description 11
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 11
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 11
- 201000000849 skin cancer Diseases 0.000 description 11
- 238000012360 testing method Methods 0.000 description 11
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 10
- 102000013462 Interleukin-12 Human genes 0.000 description 10
- 108010065805 Interleukin-12 Proteins 0.000 description 10
- 108010002616 Interleukin-5 Proteins 0.000 description 10
- 102000000743 Interleukin-5 Human genes 0.000 description 10
- 201000010099 disease Diseases 0.000 description 10
- 229960001592 paclitaxel Drugs 0.000 description 10
- 230000010287 polarization Effects 0.000 description 10
- 102000003814 Interleukin-10 Human genes 0.000 description 9
- 108090000174 Interleukin-10 Proteins 0.000 description 9
- 108010002350 Interleukin-2 Proteins 0.000 description 9
- 102000000588 Interleukin-2 Human genes 0.000 description 9
- 229930012538 Paclitaxel Natural products 0.000 description 9
- 229960000397 bevacizumab Drugs 0.000 description 9
- 230000013632 homeostatic process Effects 0.000 description 9
- 230000036039 immunity Effects 0.000 description 9
- 238000013394 immunophenotyping Methods 0.000 description 9
- 210000004698 lymphocyte Anatomy 0.000 description 9
- 108090000765 processed proteins & peptides Proteins 0.000 description 9
- 230000004083 survival effect Effects 0.000 description 9
- 102000025850 HLA-A2 Antigen Human genes 0.000 description 8
- 108010074032 HLA-A2 Antigen Proteins 0.000 description 8
- 102000003425 Tyrosinase Human genes 0.000 description 8
- 108060008724 Tyrosinase Proteins 0.000 description 8
- 210000005259 peripheral blood Anatomy 0.000 description 8
- 239000011886 peripheral blood Substances 0.000 description 8
- 238000000513 principal component analysis Methods 0.000 description 8
- 238000002271 resection Methods 0.000 description 8
- 230000000638 stimulation Effects 0.000 description 8
- 102100023688 Eotaxin Human genes 0.000 description 7
- 102100034922 T-cell surface glycoprotein CD8 alpha chain Human genes 0.000 description 7
- 108090001012 Transforming Growth Factor beta Proteins 0.000 description 7
- 102000004887 Transforming Growth Factor beta Human genes 0.000 description 7
- 238000011534 incubation Methods 0.000 description 7
- 230000003834 intracellular effect Effects 0.000 description 7
- 239000000523 sample Substances 0.000 description 7
- 238000001356 surgical procedure Methods 0.000 description 7
- 102100032367 C-C motif chemokine 5 Human genes 0.000 description 6
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 description 6
- 101000914514 Homo sapiens T-cell-specific surface glycoprotein CD28 Proteins 0.000 description 6
- 108010002335 Interleukin-9 Proteins 0.000 description 6
- 102000000585 Interleukin-9 Human genes 0.000 description 6
- 102100027213 T-cell-specific surface glycoprotein CD28 Human genes 0.000 description 6
- KQNZDYYTLMIZCT-KQPMLPITSA-N brefeldin A Chemical compound O[C@@H]1\C=C\C(=O)O[C@@H](C)CCC\C=C\[C@@H]2C[C@H](O)C[C@H]21 KQNZDYYTLMIZCT-KQPMLPITSA-N 0.000 description 6
- JUMGSHROWPPKFX-UHFFFAOYSA-N brefeldin-A Natural products CC1CCCC=CC2(C)CC(O)CC2(C)C(O)C=CC(=O)O1 JUMGSHROWPPKFX-UHFFFAOYSA-N 0.000 description 6
- 230000000875 corresponding effect Effects 0.000 description 6
- 230000004069 differentiation Effects 0.000 description 6
- 210000002865 immune cell Anatomy 0.000 description 6
- 230000001225 therapeutic effect Effects 0.000 description 6
- 102100021943 C-C motif chemokine 2 Human genes 0.000 description 5
- 101710155857 C-C motif chemokine 2 Proteins 0.000 description 5
- 102100025248 C-X-C motif chemokine 10 Human genes 0.000 description 5
- 0 CC1*CCCC1 Chemical compound CC1*CCCC1 0.000 description 5
- 101710139422 Eotaxin Proteins 0.000 description 5
- 102100039620 Granulocyte-macrophage colony-stimulating factor Human genes 0.000 description 5
- 101000946889 Homo sapiens Monocyte differentiation antigen CD14 Proteins 0.000 description 5
- 102000003812 Interleukin-15 Human genes 0.000 description 5
- 108090000172 Interleukin-15 Proteins 0.000 description 5
- 102000013691 Interleukin-17 Human genes 0.000 description 5
- 108050003558 Interleukin-17 Proteins 0.000 description 5
- 108090001005 Interleukin-6 Proteins 0.000 description 5
- 102000004889 Interleukin-6 Human genes 0.000 description 5
- 108010002586 Interleukin-7 Proteins 0.000 description 5
- 102000000704 Interleukin-7 Human genes 0.000 description 5
- 102100035877 Monocyte differentiation antigen CD14 Human genes 0.000 description 5
- 241000699666 Mus <mouse, genus> Species 0.000 description 5
- 229940126547 T-cell immunoglobulin mucin-3 Drugs 0.000 description 5
- 230000015572 biosynthetic process Effects 0.000 description 5
- 210000004970 cd4 cell Anatomy 0.000 description 5
- 238000011065 in-situ storage Methods 0.000 description 5
- 206010061289 metastatic neoplasm Diseases 0.000 description 5
- 102000004196 processed proteins & peptides Human genes 0.000 description 5
- 230000009467 reduction Effects 0.000 description 5
- 238000003786 synthesis reaction Methods 0.000 description 5
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 4
- 102100024222 B-lymphocyte antigen CD19 Human genes 0.000 description 4
- 101710098275 C-X-C motif chemokine 10 Proteins 0.000 description 4
- 108010055166 Chemokine CCL5 Proteins 0.000 description 4
- 102000019034 Chemokines Human genes 0.000 description 4
- 108010012236 Chemokines Proteins 0.000 description 4
- 101150021185 FGF gene Proteins 0.000 description 4
- 108010017080 Granulocyte Colony-Stimulating Factor Proteins 0.000 description 4
- 102000004269 Granulocyte Colony-Stimulating Factor Human genes 0.000 description 4
- 102100034458 Hepatitis A virus cellular receptor 2 Human genes 0.000 description 4
- 101710083479 Hepatitis A virus cellular receptor 2 homolog Proteins 0.000 description 4
- 241000282412 Homo Species 0.000 description 4
- 101000980825 Homo sapiens B-lymphocyte antigen CD19 Proteins 0.000 description 4
- 101000914484 Homo sapiens T-lymphocyte activation antigen CD80 Proteins 0.000 description 4
- 102100022297 Integrin alpha-X Human genes 0.000 description 4
- 108090001007 Interleukin-8 Proteins 0.000 description 4
- 102000004890 Interleukin-8 Human genes 0.000 description 4
- 102100040678 Programmed cell death protein 1 Human genes 0.000 description 4
- 101710089372 Programmed cell death protein 1 Proteins 0.000 description 4
- 102100027222 T-lymphocyte activation antigen CD80 Human genes 0.000 description 4
- BPEGJWRSRHCHSN-UHFFFAOYSA-N Temozolomide Chemical compound O=C1N(C)N=NC2=C(C(N)=O)N=CN21 BPEGJWRSRHCHSN-UHFFFAOYSA-N 0.000 description 4
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 4
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 4
- 230000001413 cellular effect Effects 0.000 description 4
- 210000004443 dendritic cell Anatomy 0.000 description 4
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 4
- 230000006870 function Effects 0.000 description 4
- 238000009396 hybridization Methods 0.000 description 4
- 230000001900 immune effect Effects 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- 238000010212 intracellular staining Methods 0.000 description 4
- 230000035800 maturation Effects 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 239000002953 phosphate buffered saline Substances 0.000 description 4
- 230000036470 plasma concentration Effects 0.000 description 4
- 230000001105 regulatory effect Effects 0.000 description 4
- 210000002966 serum Anatomy 0.000 description 4
- 229960004964 temozolomide Drugs 0.000 description 4
- 230000001988 toxicity Effects 0.000 description 4
- 231100000419 toxicity Toxicity 0.000 description 4
- 102100021663 Baculoviral IAP repeat-containing protein 5 Human genes 0.000 description 3
- 108700012434 CCL3 Proteins 0.000 description 3
- 102100035793 CD83 antigen Human genes 0.000 description 3
- 102000000013 Chemokine CCL3 Human genes 0.000 description 3
- 102000001326 Chemokine CCL4 Human genes 0.000 description 3
- 108010055165 Chemokine CCL4 Proteins 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 101000946856 Homo sapiens CD83 antigen Proteins 0.000 description 3
- 206010025652 Malignant melanoma in situ Diseases 0.000 description 3
- 210000004241 Th2 cell Anatomy 0.000 description 3
- 230000000259 anti-tumor effect Effects 0.000 description 3
- 239000012131 assay buffer Substances 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 238000001574 biopsy Methods 0.000 description 3
- 230000001684 chronic effect Effects 0.000 description 3
- 208000035250 cutaneous malignant susceptibility to 1 melanoma Diseases 0.000 description 3
- 238000007405 data analysis Methods 0.000 description 3
- 230000002349 favourable effect Effects 0.000 description 3
- 239000003102 growth factor Substances 0.000 description 3
- 102000058223 human VEGFA Human genes 0.000 description 3
- 230000028993 immune response Effects 0.000 description 3
- 239000003112 inhibitor Substances 0.000 description 3
- 102000009634 interleukin-1 receptor antagonist activity proteins Human genes 0.000 description 3
- 108040001669 interleukin-1 receptor antagonist activity proteins Proteins 0.000 description 3
- PGHMRUGBZOYCAA-ADZNBVRBSA-N ionomycin Chemical compound O1[C@H](C[C@H](O)[C@H](C)[C@H](O)[C@H](C)/C=C/C[C@@H](C)C[C@@H](C)C(/O)=C/C(=O)[C@@H](C)C[C@@H](C)C[C@@H](CCC(O)=O)C)CC[C@@]1(C)[C@@H]1O[C@](C)([C@@H](C)O)CC1 PGHMRUGBZOYCAA-ADZNBVRBSA-N 0.000 description 3
- PGHMRUGBZOYCAA-UHFFFAOYSA-N ionomycin Natural products O1C(CC(O)C(C)C(O)C(C)C=CCC(C)CC(C)C(O)=CC(=O)C(C)CC(C)CC(CCC(O)=O)C)CCC1(C)C1OC(C)(C(C)O)CC1 PGHMRUGBZOYCAA-UHFFFAOYSA-N 0.000 description 3
- 230000036210 malignancy Effects 0.000 description 3
- 108020004999 messenger RNA Proteins 0.000 description 3
- 230000001394 metastastic effect Effects 0.000 description 3
- 238000002493 microarray Methods 0.000 description 3
- 239000011325 microbead Substances 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 238000000275 quality assurance Methods 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- 108010012934 Albumin-Bound Paclitaxel Proteins 0.000 description 2
- 102100025137 Early activation antigen CD69 Human genes 0.000 description 2
- HKVAMNSJSFKALM-GKUWKFKPSA-N Everolimus Chemical compound C1C[C@@H](OCCO)[C@H](OC)C[C@@H]1C[C@@H](C)[C@H]1OC(=O)[C@@H]2CCCCN2C(=O)C(=O)[C@](O)(O2)[C@H](C)CC[C@H]2C[C@H](OC)/C(C)=C/C=C/C=C/[C@@H](C)C[C@@H](C)C(=O)[C@H](OC)[C@H](O)/C(C)=C/[C@@H](C)C(=O)C1 HKVAMNSJSFKALM-GKUWKFKPSA-N 0.000 description 2
- 101000797762 Homo sapiens C-C motif chemokine 5 Proteins 0.000 description 2
- 101000934374 Homo sapiens Early activation antigen CD69 Proteins 0.000 description 2
- 101000978392 Homo sapiens Eotaxin Proteins 0.000 description 2
- 101001057504 Homo sapiens Interferon-stimulated gene 20 kDa protein Proteins 0.000 description 2
- 101001055144 Homo sapiens Interleukin-2 receptor subunit alpha Proteins 0.000 description 2
- 101000917858 Homo sapiens Low affinity immunoglobulin gamma Fc region receptor III-A Proteins 0.000 description 2
- 101000917839 Homo sapiens Low affinity immunoglobulin gamma Fc region receptor III-B Proteins 0.000 description 2
- 102000004559 Interleukin-13 Receptors Human genes 0.000 description 2
- 108010017511 Interleukin-13 Receptors Proteins 0.000 description 2
- 102100026878 Interleukin-2 receptor subunit alpha Human genes 0.000 description 2
- 102100029185 Low affinity immunoglobulin gamma Fc region receptor III-B Human genes 0.000 description 2
- XUMBMVFBXHLACL-UHFFFAOYSA-N Melanin Chemical compound O=C1C(=O)C(C2=CNC3=C(C(C(=O)C4=C32)=O)C)=C2C4=CNC2=C1C XUMBMVFBXHLACL-UHFFFAOYSA-N 0.000 description 2
- 101100407308 Mus musculus Pdcd1lg2 gene Proteins 0.000 description 2
- 206010061309 Neoplasm progression Diseases 0.000 description 2
- 108700030875 Programmed Cell Death 1 Ligand 2 Proteins 0.000 description 2
- 102100024213 Programmed cell death 1 ligand 2 Human genes 0.000 description 2
- 238000002123 RNA extraction Methods 0.000 description 2
- 208000003837 Second Primary Neoplasms Diseases 0.000 description 2
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 2
- 108010090804 Streptavidin Proteins 0.000 description 2
- 208000036142 Viral infection Diseases 0.000 description 2
- 229940028652 abraxane Drugs 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 210000000612 antigen-presenting cell Anatomy 0.000 description 2
- 238000003491 array Methods 0.000 description 2
- 208000006673 asthma Diseases 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000000295 complement effect Effects 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 231100000673 dose–response relationship Toxicity 0.000 description 2
- 230000003828 downregulation Effects 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 229960005167 everolimus Drugs 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 230000002519 immonomodulatory effect Effects 0.000 description 2
- 230000005934 immune activation Effects 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 230000000977 initiatory effect Effects 0.000 description 2
- 229960003130 interferon gamma Drugs 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 208000037819 metastatic cancer Diseases 0.000 description 2
- 208000011575 metastatic malignant neoplasm Diseases 0.000 description 2
- 210000001616 monocyte Anatomy 0.000 description 2
- 210000004479 myeloid suppressor cell Anatomy 0.000 description 2
- 210000000822 natural killer cell Anatomy 0.000 description 2
- 230000000474 nursing effect Effects 0.000 description 2
- 239000013610 patient sample Substances 0.000 description 2
- 210000004180 plasmocyte Anatomy 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000007619 statistical method Methods 0.000 description 2
- 230000001629 suppression Effects 0.000 description 2
- 230000005751 tumor progression Effects 0.000 description 2
- 230000029069 type 2 immune response Effects 0.000 description 2
- 238000002255 vaccination Methods 0.000 description 2
- 230000009385 viral infection Effects 0.000 description 2
- 239000011534 wash buffer Substances 0.000 description 2
- RIWLPSIAFBLILR-WVNGMBSFSA-N (2s)-1-[(2s)-2-[[(2s,3s)-2-[[(2s)-2-[[(2s,3r)-2-[[(2r,3s)-2-[[(2s)-2-[[2-[[2-[acetyl(methyl)amino]acetyl]amino]acetyl]amino]-3-methylbutanoyl]amino]-3-methylpentanoyl]amino]-3-hydroxybutanoyl]amino]pentanoyl]amino]-3-methylpentanoyl]amino]-5-(diaminomethy Chemical compound CC(=O)N(C)CC(=O)NCC(=O)N[C@@H](C(C)C)C(=O)N[C@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N1CCC[C@H]1C(=O)NCC RIWLPSIAFBLILR-WVNGMBSFSA-N 0.000 description 1
- 108010058566 130-nm albumin-bound paclitaxel Proteins 0.000 description 1
- 244000118350 Andrographis paniculata Species 0.000 description 1
- 102100022005 B-lymphocyte antigen CD20 Human genes 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 210000004366 CD4-positive T-lymphocyte Anatomy 0.000 description 1
- 108010029697 CD40 Ligand Proteins 0.000 description 1
- 102100032937 CD40 ligand Human genes 0.000 description 1
- 210000001266 CD8-positive T-lymphocyte Anatomy 0.000 description 1
- 229940045513 CTLA4 antagonist Drugs 0.000 description 1
- 241000282472 Canis lupus familiaris Species 0.000 description 1
- 241000282693 Cercopithecidae Species 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- CMSMOCZEIVJLDB-UHFFFAOYSA-N Cyclophosphamide Chemical compound ClCCN(CCCl)P1(=O)NCCCO1 CMSMOCZEIVJLDB-UHFFFAOYSA-N 0.000 description 1
- 208000000471 Dysplastic Nevus Syndrome Diseases 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 241000283086 Equidae Species 0.000 description 1
- 241000282326 Felis catus Species 0.000 description 1
- 102000004457 Granulocyte-Macrophage Colony-Stimulating Factor Human genes 0.000 description 1
- 102000006354 HLA-DR Antigens Human genes 0.000 description 1
- 108010058597 HLA-DR Antigens Proteins 0.000 description 1
- 102000001554 Hemoglobins Human genes 0.000 description 1
- 108010054147 Hemoglobins Proteins 0.000 description 1
- 101000897405 Homo sapiens B-lymphocyte antigen CD20 Proteins 0.000 description 1
- 101000858088 Homo sapiens C-X-C motif chemokine 10 Proteins 0.000 description 1
- 101000599940 Homo sapiens Interferon gamma Proteins 0.000 description 1
- 101001002709 Homo sapiens Interleukin-4 Proteins 0.000 description 1
- 101001018097 Homo sapiens L-selectin Proteins 0.000 description 1
- 101000581981 Homo sapiens Neural cell adhesion molecule 1 Proteins 0.000 description 1
- 101001094545 Homo sapiens Retrotransposon-like protein 1 Proteins 0.000 description 1
- 101000742596 Homo sapiens Vascular endothelial growth factor C Proteins 0.000 description 1
- 108010047761 Interferon-alpha Proteins 0.000 description 1
- 102000006992 Interferon-alpha Human genes 0.000 description 1
- 102100033467 L-selectin Human genes 0.000 description 1
- 238000012773 Laboratory assay Methods 0.000 description 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- 241000161982 Mogera robusta Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 108010072915 NAc-Sar-Gly-Val-(d-allo-Ile)-Thr-Nva-Ile-Arg-ProNEt Proteins 0.000 description 1
- 102100027347 Neural cell adhesion molecule 1 Human genes 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 206010033128 Ovarian cancer Diseases 0.000 description 1
- 206010061535 Ovarian neoplasm Diseases 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- 208000037581 Persistent Infection Diseases 0.000 description 1
- 206010060862 Prostate cancer Diseases 0.000 description 1
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 1
- 238000010802 RNA extraction kit Methods 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 238000000692 Student's t-test Methods 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- 206010042496 Sunburn Diseases 0.000 description 1
- 230000024932 T cell mediated immunity Effects 0.000 description 1
- 230000006052 T cell proliferation Effects 0.000 description 1
- 210000000662 T-lymphocyte subset Anatomy 0.000 description 1
- 108700012920 TNF Proteins 0.000 description 1
- 102000013530 TOR Serine-Threonine Kinases Human genes 0.000 description 1
- 108010065917 TOR Serine-Threonine Kinases Proteins 0.000 description 1
- 210000000447 Th1 cell Anatomy 0.000 description 1
- 108010046722 Thrombospondin 1 Proteins 0.000 description 1
- 102100036034 Thrombospondin-1 Human genes 0.000 description 1
- 108010073929 Vascular Endothelial Growth Factor A Proteins 0.000 description 1
- 102000009524 Vascular Endothelial Growth Factor A Human genes 0.000 description 1
- 102100038232 Vascular endothelial growth factor C Human genes 0.000 description 1
- 208000038016 acute inflammation Diseases 0.000 description 1
- 230000006022 acute inflammation Effects 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 230000000735 allogeneic effect Effects 0.000 description 1
- 239000002269 analeptic agent Substances 0.000 description 1
- 230000001772 anti-angiogenic effect Effects 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000005809 anti-tumor immunity Effects 0.000 description 1
- 230000005975 antitumor immune response Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 238000001815 biotherapy Methods 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 244000309466 calf Species 0.000 description 1
- 230000005773 cancer-related death Effects 0.000 description 1
- JJWKPURADFRFRB-UHFFFAOYSA-N carbonyl sulfide Chemical compound O=C=S JJWKPURADFRFRB-UHFFFAOYSA-N 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 230000007969 cellular immunity Effects 0.000 description 1
- 230000008614 cellular interaction Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000012829 chemotherapy agent Substances 0.000 description 1
- 238000003501 co-culture Methods 0.000 description 1
- 210000001072 colon Anatomy 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000000139 costimulatory effect Effects 0.000 description 1
- 229960004397 cyclophosphamide Drugs 0.000 description 1
- 230000016396 cytokine production Effects 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 231100000517 death Toxicity 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000013024 dilution buffer Substances 0.000 description 1
- 230000003292 diminished effect Effects 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 230000002222 downregulating effect Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 238000010195 expression analysis Methods 0.000 description 1
- 210000003754 fetus Anatomy 0.000 description 1
- 239000012997 ficoll-paque Substances 0.000 description 1
- 235000013312 flour Nutrition 0.000 description 1
- 238000010230 functional analysis Methods 0.000 description 1
- 210000002443 helper t lymphocyte Anatomy 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 229940094991 herring sperm dna Drugs 0.000 description 1
- 230000002962 histologic effect Effects 0.000 description 1
- 102000055229 human IL4 Human genes 0.000 description 1
- 230000009390 immune abnormality Effects 0.000 description 1
- 230000008102 immune modulation Effects 0.000 description 1
- 230000006058 immune tolerance Effects 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 230000002055 immunohistochemical effect Effects 0.000 description 1
- 238000003364 immunohistochemistry Methods 0.000 description 1
- 238000013388 immunohistochemistry analysis Methods 0.000 description 1
- 238000012744 immunostaining Methods 0.000 description 1
- 238000002650 immunosuppressive therapy Methods 0.000 description 1
- 239000007943 implant Substances 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000010348 incorporation Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000018711 interleukin-13 production Effects 0.000 description 1
- 238000010255 intramuscular injection Methods 0.000 description 1
- 239000007927 intramuscular injection Substances 0.000 description 1
- 238000007913 intrathecal administration Methods 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 201000005202 lung cancer Diseases 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 230000000527 lymphocytic effect Effects 0.000 description 1
- 238000011469 lymphodepleting chemotherapy Methods 0.000 description 1
- 238000010841 mRNA extraction Methods 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 210000002752 melanocyte Anatomy 0.000 description 1
- 238000001471 micro-filtration Methods 0.000 description 1
- 239000004005 microsphere Substances 0.000 description 1
- 238000007799 mixed lymphocyte reaction assay Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000003068 molecular probe Substances 0.000 description 1
- 238000007837 multiplex assay Methods 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 230000001338 necrotic effect Effects 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 208000004649 neutrophil actin dysfunction Diseases 0.000 description 1
- 238000011275 oncology therapy Methods 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 230000005298 paramagnetic effect Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 238000009521 phase II clinical trial Methods 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 239000006187 pill Substances 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 230000035935 pregnancy Effects 0.000 description 1
- 238000009597 pregnancy test Methods 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 239000001397 quillaja saponaria molina bark Substances 0.000 description 1
- 210000000664 rectum Anatomy 0.000 description 1
- 210000003289 regulatory T cell Anatomy 0.000 description 1
- 230000004043 responsiveness Effects 0.000 description 1
- 229930182490 saponin Natural products 0.000 description 1
- 150000007949 saponins Chemical class 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000013207 serial dilution Methods 0.000 description 1
- 230000001568 sexual effect Effects 0.000 description 1
- 210000003491 skin Anatomy 0.000 description 1
- 210000000329 smooth muscle myocyte Anatomy 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 238000011301 standard therapy Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 230000036561 sun exposure Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 238000000844 transformation Methods 0.000 description 1
- 238000013520 translational research Methods 0.000 description 1
- 238000011269 treatment regimen Methods 0.000 description 1
- 210000003171 tumor-infiltrating lymphocyte Anatomy 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
- 239000002525 vasculotropin inhibitor Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/22—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against growth factors ; against growth regulators
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/337—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having four-membered rings, e.g. taxol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/4164—1,3-Diazoles
- A61K31/4188—1,3-Diazoles condensed with other heterocyclic ring systems, e.g. biotin, sorbinil
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/555—Heterocyclic compounds containing heavy metals, e.g. hemin, hematin, melarsoprol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/66—Phosphorus compounds
- A61K31/675—Phosphorus compounds having nitrogen as a ring hetero atom, e.g. pyridoxal phosphate
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/0005—Vertebrate antigens
- A61K39/0011—Cancer antigens
- A61K39/001148—Regulators of development
- A61K39/00115—Apoptosis related proteins, e.g. survivin or livin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/0005—Vertebrate antigens
- A61K39/0011—Cancer antigens
- A61K39/001154—Enzymes
- A61K39/001156—Tyrosinase and tyrosinase related proteinases [TRP-1 or TRP-2]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/0005—Vertebrate antigens
- A61K39/0011—Cancer antigens
- A61K39/00119—Melanoma antigens
- A61K39/001191—Melan-A/MART
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/0005—Vertebrate antigens
- A61K39/0011—Cancer antigens
- A61K39/00119—Melanoma antigens
- A61K39/001192—Glycoprotein 100 [Gp100]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
- A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
- A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
- A61K39/3955—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against proteinaceous materials, e.g. enzymes, hormones, lymphokines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/545—Medicinal preparations containing antigens or antibodies characterised by the dose, timing or administration schedule
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/80—Vaccine for a specifically defined cancer
- A61K2039/876—Skin, melanoma
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
- A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
- A61K51/04—Organic compounds
- A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins
- A61K51/10—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody
- A61K51/1045—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody against animal or human tumor cells or tumor cell determinants
- A61K51/1069—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody against animal or human tumor cells or tumor cell determinants the tumor cell being from blood cells, e.g. the cancer being a myeloma
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
- C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
Definitions
- This document relates to methods and materials involved in treating cancer (e.g., melanoma).
- cancer e.g., melanoma
- this document relates to methods and materials involved in using an anti-chronic inflammation treatment (e.g., chemotherapy) in combination with a cancer treatment agent (e.g., a cancer vaccine) to treat cancer.
- an anti-chronic inflammation treatment e.g., chemotherapy
- a cancer treatment agent e.g., a cancer vaccine
- Melanoma is the most serious form of skin cancer. It is a malignant tumor that originates in melanocytes, the cells which produce the pigment melanin that colors skin, hair, and eyes and is heavily concentrated in most moles. While it is not the most common type of skin cancer, melanoma underlies the majority of skin cancer-related deaths. About 48,000 deaths worldwide are registered annually as being due to malignant melanoma. Worldwide, there are about 160,000 new cases of melanoma each year. Melanoma is more frequent in white men and is particularly common in white populations living in sunny climates. Other risk factors for developing melanoma include a history of sunburn, excessive sun exposure, living in a sunny climate or at high altitude, having many moles or large moles, and a family or personal history of skin cancer.
- this document provides methods and materials related to treating cancer.
- this document provides methods and materials for using an anti-chronic inflammation treatment (e.g., chemotherapy) in combination with a cancer treatment agent (e.g., a cancer vaccine) to treat cancer.
- a cancer treatment agent e.g., a cancer vaccine
- cancer can induce a global state of immune dysfunction and/or chronic inflammation in the cancer patient.
- This global state of immune dysfunction and/or chronic inflammation can prevent the patient from mounting a successful response against the cancer.
- a cancer patient with a global state of chronic inflammation can be in a state such that the patient is unable to generate an anti-cancer immune response when given an anti-cancer vaccine.
- the methods and materials provided herein can be used to reduce the global state of immune dysfunction and/or chronic inflammation present within a cancer patient such that the cancer patient can better respond to a cancer treatment such as a cancer vaccine.
- a cancer treatment such as a cancer vaccine.
- chemotherapy, radiation, anti-IL-4 agents (e.g., anti-IL-4 antibodies), anti-IL-13 agents (e.g., soluble IL-13 receptor), steroids, and combinations thereof can be used to reduce the global state of immune dysfunction and/or chronic inflammation present within a cancer patient.
- an appropriate cancer treatment such as a cancer vaccine or other immune stimulating agents (e.g., IL-2 or IL-12).
- this document features a method for treating a mammal having cancer.
- the method comprises, or consists essentially of, (a) administering to the mammal an anti-chronic inflammation treatment under conditions wherein the level of global chronic inflammation in the mammal is reduced, and (b) administering to the mammal a cancer treatment agent under conditions wherein the presence of the cancer is reduced.
- the mammal can be a human.
- the cancer can be melanoma.
- the cancer can be stage IV melanoma.
- the anti-chronic inflammation treatment can comprise chemotherapy, radiation, an anti-IL-4 agent, an anti-IL-13 agent, or a steroid treatment.
- the cancer treatment agent can be a cancer vaccine.
- the cancer vaccine can be a MART-1, gp100, or survivin cancer vaccine.
- the period of time between the last administration of the anti-chronic inflammation treatment and the first administration of the cancer treatment agent can be between two weeks and six months.
- this document features a method for treating a mammal having cancer.
- the method comprises, or consists essentially of, (a) administering to the mammal an anti-TGF ⁇ antibody under conditions wherein the level of global chronic inflammation in the mammal is reduced, and (b) administering to the mammal a cancer treatment agent under conditions wherein the presence of the cancer is reduced.
- the mammal can be a human.
- the cancer can be melanoma.
- the cancer can be stage IV melanoma.
- the cancer treatment agent can be a cancer vaccine.
- the cancer vaccine can be a MART-1, gp100, or survivin cancer vaccine.
- the period of time between the last administration of the anti-TGF ⁇ antibody and the first administration of the cancer treatment agent can be between two weeks and six months.
- FIG. 1 Effects of 1% patient plasma (vs. normal plasma) on in vitro maturation of normal DC. Presented is a representative experiment demonstrating the % co-stimulatory molecule positive (CD80, 83, 86) DC.
- FIG. 2 Allogeneic mixed lymphocyte reaction cultures evaluating proliferation of T cells mixed at differing ratios with mature or immature dendritic cells in the presence of normal (control) plasma or plasma from a patient with metastatic melanoma (MTB plasma). T cell proliferation was assessed by 3H-TdR incorporation. Similar results were observed in two other experiments.
- FIG. 3 Three dimensional representation of the results of Principal Component Analysis (PCA).
- PCA Principal Component Analysis
- the PCA was performed on 234 clinical samples for concentrations of 27 cytokines. Each sphere represents one clinical sample. The dark spheres represent stage IV melanoma patients, and the lighter spheres represent atypical nevi, benign nevi, in situ melanoma, stage I melanoma, stage II melanoma, or stage III melanoma. The axes are the main principal components. Significant grouping is only evident for patients in the cohort of stage IV (metastatic) melanoma (dark spheres).
- FIG. 4 Mean plasma IL-4 levels (pg/mL) ⁇ SD across stages of melanoma (melanoma in situ and stages: I, II, III, IV), patients with atypical nevi (atypical) and benign nevi (normal controls).
- FIG. 5 Assessment of T-cell phenotype and function in healthy donors and stage IV melanoma patients.
- the number of T-cells exhibiting the FoxP3 (Treg) or PD-1 phenotype in peripheral blood was determined in healthy donors and stage IV melanoma patients (A).
- the frequency of FoxP3 positive cells were measured by 3-color flow cytometry, CD4-PC5, CD25-PE and FoxP3-Alexa flour 488.
- the mean percent (+/ ⁇ SD) of FoxP3 positive were determined from the CD4 and CD25 double positive population.
- the mean frequency (+/ ⁇ SD) of PD-1+ cells was measured from the CD8+ population.
- the frequency (+/ ⁇ SD) of tetramer positive (CMV or MART-1) CD8+ T cells was compared among normal volunteers and patients with stage IV melanoma (B).
- FIG. 6 VEGF levels in patients with metastatic melanoma.
- A RNA expression of cytokines in human metastatic melanoma tissue. Twenty-four frozen biopsies of metastatic melanoma tumor tissues was used to extract RNA for expression array analysis. Illustrated are the RNA expression intensity profiles of 45 probes for 24 cytokines.
- FIG. 7 Healthy donor PBMCs were cultured in vitro for 48 hours in the presence of increasing concentrations of recombinant human VEGFA. At the end of the incubation, cells were stimulated with PMA and ionomycin, in the presence of brefeldin A and stained for intracellular IFN ⁇ , IL-4, or IL-13 and surface immunophenotyped for CD294 or TIM-3. Cells were then analyzed for the frequency of CD4 cells (% of CD4) expressing said phenotypes using flow cytometry.
- FIG. 8 Co-culture with recombinant human VEGF shifts T-helper polarity from Th1 (IFN- ⁇ ) to Th2 (IL-4) predominance.
- PBMC isolated from healthy donors were stimulated with PMA and ionomycin in the presence of brefeldin-A, permeabolized, and intracellularly stained for human IFN- ⁇ (FITC) and human IL-4 (PE).
- FITC human IFN- ⁇
- PE human IL-4
- PBMC were exposed to increasing concentrations of VEGF (0-16 pg/mL) without/with IL-12. All cells were immunostained with PC5 anti-human CD4.
- Purified CD4+ T-cells B were negatively isolated using Miltenyi beads, cultured, and stained in the same fashion as PBMC (A). Similar results were observed in 5 different experiments.
- FIG. 9 Changes in plasma VEGF levels (plasma VEGFA in pg/mL; top left) at three time points in a single patient with metastatic melanoma treated on protocol N047a correlate with improved Th1/Th2 ratio as determined by intracellular staining of CD4+ cells for IFN gamma or IL-4 (top right). These also correlate with emergence of increased frequencies of tumor specific CTL (bottom right) as determined by tetramer assay.
- FIG. 10 Cellular interactions of acute and chronic inflammation.
- MSC myeloid suppressor cell
- Th1 & Th2 T helper lymphocytes type 1 & 2
- Treg regulatory T cell
- DC1 dendritic cells, type 1
- DC2 dendritic cells type 2
- CTL cytotoxic T lymphocyte
- FIG. 11 Correlation of surface immunophenotyping for CD294 and TIM-3 with intracellular IL-4, IL-13 and IFN ⁇ for the purposes of enumeration of Th2 and Th1 cells respectively.
- FIG. 12 Changes in the ratio of human PBMC derived CD4 T cell subsets (Th1 vs. Th2) following in vitro incubation with varying concentrations of VEGFA or TGF ⁇ .
- FIG. 13 Relative ratios of human PBMC derived CD4 T cells subsets (Th1 vs. Th2) cultured in vitro with varying concentrations of VEGFA in the absence or presence of increasing concentrations of anti-TGF ⁇ neutralizing antibody. Untreated (media) as well as Th1 (Th1) and Th2 (Th2) favorable in vitro conditions are presented as controls. These results indicate that presence of anti-TGF ⁇ antibodies reverses the Th1/Th2 modulation of VEGFA in vitro, suggesting that the observed VEGF effect in these cells may be TGF ⁇ mediated.
- This document provides methods and materials related to treating cancer in mammals.
- this document provides methods and materials related to the use of an anti-chronic inflammation treatment (e.g., chemotherapy) in combination with a cancer treatment agent (e.g., a cancer vaccine) to treat cancer.
- an anti-chronic inflammation treatment e.g., chemotherapy
- a cancer treatment agent e.g., a cancer vaccine
- the methods and materials provided herein can be used to treat cancer in any type of mammal including, without limitation, mice, rats, dogs, cats, horses, cows, pigs, monkeys, and humans.
- Any type of cancer such as skin cancer (e.g., melanoma), can be treated.
- Examples of cancer that can be treated as described herein include, without limitation, skin cancer, lung cancer, breast cancer, prostate cancer, ovarian cancer, and colon cancer.
- stage I, stage II, stage III, or stage IV melanoma can be treated using the methods and materials provided herein.
- cancer can be treated by administering an anti-chronic inflammation treatment such that the global state of immune dysfunction and/or chronic inflammation present within a cancer patient is reduced.
- an anti-chronic inflammation treatment such that the global state of immune dysfunction and/or chronic inflammation present within a cancer patient is reduced.
- chemotherapy radiation, anti-IL-4 agents (e.g., anti-IL-4 antibodies), anti-IL-13 agents (e.g., soluble IL-13 receptor), steroids, and combinations thereof can be used to reduce the global state of immune dysfunction and/or chronic inflammation present within a cancer patient.
- chemotherapy such as paclitaxel, carboplatin, temozolomide, or cyclophosphamide can be administered to a cancer patient to reduce the global state of immune dysfunction and/or chronic inflammation present within a cancer patient.
- an anti-chronic inflammation treatment can include, without limitation, administering an anti-TGF ⁇ antibody.
- anti-TGF ⁇ antibodies can be administered to a cancer patient to reduce the global state of immune dysfunction and/or chronic inflammation present within a cancer patient.
- anti-TGF ⁇ antibodies include, without limitation, human monoclonal anti-TGF- ⁇ 1 antibodies such as CAT-192 (Genzyme Inc.).
- cytokine profiles e.g., IL-4, IL-13, IL-4, IL-13, IL-5, IL-10, IL-2, and interferon gamma
- cytokine profiles e.g., IL-4, IL-13, IL-4, IL-13, IL-5, IL-10, IL-2, and interferon gamma
- the cancer patient can be treated with an appropriate cancer treatment (e.g., an immune cancer treatment) such as a cancer vaccine.
- appropriate cancer treatment agents include, without limitation, immune stimulating cytokines (e.g., IL-2, IL-12, interferon alpha, and interferon gamma), inhibitors of immune down-regulation (e.g., anti-CTLA4, anti-41bb, anti-PD-1, and anti-CD25), and cancer vaccines (e.g., MART-1, gp100, survivin, and tyrosinase cancer vaccines).
- paclitaxel, carboplatin, bevacizumab, and anti-CTLA-4 can be used to treat (e.g., skin cancer) upon administration either individually or in any combination thereof (e.g., paclitaxel, carboplatin and bevacizumab).
- the amount of time between administration of an anti-chronic inflammation treatment and administration of a cancer treatment can be between two weeks and twelve months (e.g., between two weeks and eleven months, between two weeks and ten months, between two weeks and nine months, between two weeks and eight months, between two weeks and seven months, between two weeks and six months, between one month and twelve months, between one month and six months, or between two months and six months).
- a chemotherapy agent e.g., paclitaxel
- a cancer treatment e.g., a cancer vaccine
- a cancer treatment agent can be administered orally or via injection (e.g., subcutaneous injection, intramuscular injection, intravenous injection, or intrathecal injection).
- injection e.g., subcutaneous injection, intramuscular injection, intravenous injection, or intrathecal injection.
- cancer treatment agents can be administered by different routes.
- one cancer treatment agent can be administered orally and a second cancer treatment agent can be administered via injection.
- a cancer treatment agent can be administered following resection of a tumor.
- Cancer treatment agent can be administered to a mammal in any amount, at any frequency, and for any duration effective to achieve a desired outcome (e.g., to increase progression-free survival or to increase the time to progression).
- cancer treatment agents can be administered to a mammal having skin cancer to reduce the progression rate of melanoma by 5, 10, 25, 50, 75, 100, or more percent.
- the progression rate can be reduced such that no additional cancer progression is detected.
- Any method can be used to determine whether or not the progression rate of skin cancer is reduced.
- the progression rate of skin cancer can be assessed by imaging tissue at different time points and determining the amount of cancer cells present.
- the amounts of cancer cells determined within tissue at different times can be compared to determine the progression rate. After treatment as described herein, the progression rate can be determined again over another time interval. In some cases, the stage of skin cancer after treatment can be determined and compared to the stage before treatment to determine whether or not the progression rate was reduced.
- a cancer treatment agent can be administered to a mammal having cancer under conditions where progression-free survival or time to progression is increased (e.g., by 5, 10, 25, 50, 75, 100, or more percent) as compared to the median progression-free survival or time to progression, respectively, of corresponding mammals having untreated cancer.
- An effective amount of a cancer treatment agent can be any amount that reduces the progression rate of cancer, increases the progression-free survival rate, or increases the median time to progression without producing significant toxicity to the mammal
- an effective amount of a cancer treatment agent such as bevacizumab can be from about 5 mg/kg/week to about 15 mg/kg/week (e.g., about 10 mg/kg/week). If a particular mammal fails to respond to a particular amount, then the amount of one or more of the compounds can be increased by, for example, two fold. After receiving this higher concentration, the mammal can be monitored for both responsiveness to the treatment and toxicity symptoms, and adjustments made accordingly.
- the effective amount can remain constant or can be adjusted as a sliding scale or variable dose depending on the mammal's response to treatment.
- Various factors can influence the actual effective amount used for a particular application. For example, the frequency of administration, duration of treatment, use of multiple treatment agents, route of administration, and severity of the cancer may require an increase or decrease in the actual effective amount administered.
- the frequency of administration can be any frequency that reduces the progression rate of cancer, increases the progression-free survival rate, or increases the median time to progression without producing significant toxicity to the mammal
- the frequency of administration can be from about once a month to about three times a month, or from about twice a month to about six times a month, or from about once every two months to about three times every two months.
- the frequency of administration can remain constant or can be variable during the duration of treatment.
- the frequency of administration of multiple cancer treatment agents can be the same or can differ. For example, one cancer treatment agent can be administered three times during a 28 day period, while a second cancer treatment agent can be administered one time, and third cancer treatment agent can be administered two times during the same period.
- a course of treatment with a cancer treatment agent can include rest periods.
- a cancer treatment agent can be administered over a two week period followed by a two week rest period, and such a regimen can be repeated multiple times.
- various factors can influence the actual frequency of administration used for a particular application. For example, the effective amount, duration of treatment, use of multiple treatment agents, route of administration, and severity of the cancer may require an increase or decrease in administration frequency.
- an effective duration for administering a composition provided herein can be any duration that reduces the progression rate of cancer, increases the progression-free survival rate, or increases the median time to progression without producing significant toxicity to the mammal
- the effective duration can vary from several days to several weeks, months, or years.
- the effective duration for the treatment of cancer can range in duration from several weeks to several months.
- an effective duration can be for as long as an individual mammal is alive. Multiple factors can influence the actual effective duration used for a particular treatment.
- an effective duration can vary with the frequency of administration, effective amount, use of multiple treatment agents, route of administration, and severity of the cancer.
- the mammal After administering a composition provided herein to a mammal, the mammal can be monitored to determine whether or not the cancer was treated. For example, a mammal can be assessed after treatment to determine whether or not the progression rate of cancer was reduced (e.g., stopped). As described herein, any appropriate method can be used to assess progression and survival rates.
- Peripheral venous blood 50-90 mL was drawn into heparinized Vacutainer tubes that were processed and separated into plasma and peripheral blood mononuclear cells (PBMC) following gradient centrifugation using Ficoll-Paque (GE Healthcare Uppsala, Sweden). Plasma was collected and immediately frozen at ⁇ 80° C. (1 mL aliquots). PBMC were collected, washed in phosphate buffered saline (PBS), counted, diluted to 1 ⁇ 10 7 /mL and viably frozen in 90% cosmic calf serum (Hyclone Inc. Logan, Utah) and 10% DMSO (Sigma St. Louis, Mo.). All assays were batch-analyzed at the end of the study.
- PBS phosphate buffered saline
- anti-human monoclonal antibodies were used in PBMC immunophenotyping for flow cytometry: anti-CD3-APC, FITC and PE, anti-CD4-FITC, anti-CD8-PE, anti-CD16 PE, anti-CD56 PE, anti-CD62L APC, anti-CD69 FITC, anti-CD14 FITC, anti-CD16 FITC, anti-CD19 FITC, anti-CD11c APC, anti-CD80 PE, anti-CD83 PE, anti-CD86 PE, anti-CD40 APC, anti-HLA-DR PC5, anti-PD-1 (BD Pharmingen San Jose, Calif.).
- the human monoclonal antibodies anti-CD4 PC5 and anti-CD25 PE were purchased from Biolegend (San Diego, Calif.) and used in conjunction with anti-human FoxP3 for the enumeration of T reg cells.
- the following anti-human monoclonal antibodies were used for intracellular staining for flow cytometry: anti-IFN ⁇ FITC, anti-IL-13 PE, anti-IL-4 PE (R and D Systems Minneapolis, Minn.), and anti-FoxP3 Alexaflour 488 (Biolegend San Diego, Calif.).
- PBMC Previously frozen PBMC (0.5-1.0 ⁇ 10 6 cells/mL) were thawed and aliquoted into 96 well rounded bottom plates (100 ⁇ L/well). The desired antibody or antibody pool was added at 5 ⁇ L/well. The cells and antibodies were incubated for 30 minutes at 4° C. and washed twice with 1 ⁇ PBS (Cellgro Manassas, Va.), 0.1% BSA and 0.05% sodium azide (Sigma St. Louis, Mo.). Four-color flow cytometry was performed on a LSRII flow cytometer (Becton Dickenson San Jose, Calif.), and Cellquest software (Becton Dickenson San Jose, Calif.) was utilized for data analysis.
- PBS Cellgro Manassas, Va.
- BSA 0.1% BSA
- sodium azide Sigma St. Louis, Mo.
- a gate was set on cells, which were HLA-DR + and Lin ⁇ (CD3, CD14, CD16 and CD19). From this population the percentage of cells, which were CD11c + and positive for costimulatory molecules (CD80, CD83 and CD86) was determined as previously elsewhere (Fricke et al., Clin. Cancer Res., 13:4840-8 (2007)).
- a panel of tumor associated antigen tetramers, MART-1 26-35 , gp100 264-272 , gp100 209-217 , and tyrosinase 369-377 were used to enumerate the frequency of tumor antigen specific CD8 positive T-cells.
- EBV 280-288 and CMV 495-503 were used as positive controls.
- a gate was set on lymphocytes, which were CD8 + and negative for CD4, CD14 and CD 19.
- Three-color flow cytometry was performed on a LSRII flow cytometer (Becton Dickenson San Jose, Calif.) and Cellquest software (Becton Dickenson San Jose, Calif.) was utilized for data analysis.
- aAPC artificial antigen presenting cell method
- Protein levels for 27 cytokines, chemokines, and growth factors including IL-1 ⁇ , IL-1ra, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12p70, IL-13, IL-15, IL-17, Eotaxin, FGF basic, G-CSF, GM-CSF, IFN- ⁇ , IP-10, MCP-1, MIP-1 ⁇ , MIP-1 ⁇ , PDGF, RANTES, TNF- ⁇ , and VEGF, were measured using the Bio-plex cytokine assay (Bio-rad, Hercules, Calif.) as per manufacturer's instructions.
- Patient plasma was diluted 1:4 in dilution buffer and 50 ⁇ L was added to washed, fluorescently dyed microspheres (beads) to which biomolecules of interest are bound.
- the beads and diluted patient plasma were incubated for 30 minutes at room temperature with agitation. After the incubation the beads were washed in Bio-plex wash buffer and placed in 25 ⁇ L of detection antibody and incubated for 30 minutes as described above. After washing, the beads were placed in streptavidin-PE, incubated, and washed a final time.
- the bound beads were resuspended in 125 ⁇ L Bio-plex assay buffer and read with the Luminex plate reader (Bio-rad, Hercules, Calif.). Protein concentrations were determined using a standard curve generated using the high PMT concentrations with sensitivity from 10-1000 pg/mL.
- PBMC from healthy donors were stimulated for 3 days with CD3/CD28 expander beads (Invitrogen Oslo, Norway) with and without increasing doses of recombinant VEGF (1-16 pg/mL).
- Cells were also cultured with 10 ⁇ g/mL recombinant human IL-12 (R and D Systems, Minneapolis, Minn.) or 8 ⁇ g/mL of a monoclonal anti-human IL-12 (R and D Systems Minneapolis, Minn. clone #24910). After the culture, the cells were harvested and restimulated with 50 ng/mL PMA (Sigma, St.
- RNA transcript labeling reagent Affymetrix, Santa Clara, Calif.
- Labeled cRNA was then fragmented and hybridized onto the U133 Plus 2.0 array.
- Appropriate amounts of fragmented cRNA and control oligonucleotide B2 were added along with control cRNA (BioB, BioC, and BioD), herring sperm DNA, and bovine serum albumin to the hybridization buffer.
- the hybridization mixture was heated at 99° C. for 5 minutes followed by incubation at 45° C. for 5 minutes before injecting the sample into the microarray. Then, the hybridization was carried out at 45° C. for 16 hours with mixing on a rotisserie at 60 rpm. After hybridization, the solutions were removed, and the arrays were washed and then stained with streptavidin-phycoerythrin (Molecular Probes, Eugene, Oreg.). After washes, arrays were scanned using the GeneChip Scanner 3000 (Affymetrix, Santa Clara, Calif.).
- the quality of the fragmented biotin labeled cRNA in each experiment was evaluated before hybridizing onto the U133A expression array by both obtaining electropherograms on Agilent 2100 Bioanalyzer and hybridizing a fraction of the sample onto test-3 array as a measure of quality control.
- GeneSpring GX 7.3 (Agilent Technologies, Inc. Santa Clara, Calif.) data analysis software was used to analyze the results of the microarray experiment. Gene expression values were normalized by the GCRMA algorithm (Bolstad et al., Bioinformatics, 19:185-93 (2003)).
- PCA principal component analysis
- the identity of the unknown factor(s) could be a known cytokine.
- a screening study was performed to quantify the plasma concentrations of 27 different cytokines (BioRad human 27-plex cytokine panel assaying for plasma concentrations of IL-1 ⁇ , IL-1r ⁇ , IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12 (p70), IL-13, IL-15, IL-17, basic FGF, eotaxin, G-CSF, GM-CSF, IFN- ⁇ , IP-10, MCP-1, MIP-1 ⁇ , MIP-1 ⁇ , PDGF, RANTES, TNF- ⁇ , and VEGF) in plasma of over 200 patients with all stages of melanoma (stage I thought IV), melanoma in situ, atypical nevi (possible pre-malignant lesions) as well as normal controls (patients with benign nevi).
- PCA principal component analysis
- PBMC isolated from patients with benign nevi, atypical (including dysplastic) nevi, as well as patients with in situ, stage I, II, III or IV melanoma were analyzed by flow cytometry to determine the frequencies of T, NK, and dendritic (DC) cell subsets.
- T, NK, and dendritic (DC) cell subsets There were not significant differences in frequencies of T-cell among stages of melanoma as determined by numbers of CD3, CD4 or CD8 positive T-cells (Table 2).
- no significant differences were found in activated T-cells (CD3/CD69), total NK cells (CD16/56 + , CD3 ⁇ ), or most DC subset parameters.
- stage IV melanoma As patients with stage IV melanoma appeared to differ significantly from all others with regard to plasma cytokine profiles, the cell subset analysis of patients with stage IV melanoma were compared relative to all others. The analysis revealed no significant differences among most parameters (Table 3) with the following exceptions: (a) the frequency of na ⁇ ve T-cells (CD3/CD62L + ) as well as activated DC (CD11c/CD83 + ) were significantly less in patients with stage IV melanoma; and (b) the frequency of tetramer positive CTL for gp100 and tyrosinase (but not MART-1 or CMV and EBV) were increased in patients with stage IV melanoma.
- T h1 and T h2 enumeration could not be performed. These data suggested that there appeared to be some level of “immune activation” in patients with metastatic melanoma that was different from all other cohorts, and this was consistent with a state of T h2 mediated “chronic inflammation.”
- stage 4 disease The plasma cytokine profiling data comparing patients across all stages of melanoma suggested that the greatest differences in the measured parameters occurred in the setting of metastatic melanoma (stage 4 disease). Therefore, it was hypothesized, that the presence of visible metastatic disease was in some way responsible for the detected Th 2 cytokine dominance in these patients and was likely the result of molecules produced by the tumor and/or its interaction with surrounding immune cells. To that end, the mRNA expression profile of 24 biopsy specimens of human metastatic melanoma was analyzed looking for up-regulation of expression of known regulatory molecules of immunity (cytokines and chemokines).
- cytokines and chemokines known regulatory molecules of immunity
- the mRNA was extracted from frozen sections in areas that by H&E staining appeared to contain pure tumor tissue (devoid of necrotic tissue, stroma or lymphocytic infiltrates).
- the RNA was analyzed using an Affymetrix U133 plus 2.0 array. In this experiment, concurrent blood samples were not available from the patients for whom tumor tissues existed.
- cytokines 45 probes: IL1a and b, IL-1ra, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12, IL-13, IL-15, IL-17, IFN- ⁇ , CCL5, CCL11, CSF-2, MCP-1, TNF- ⁇ , VEGF was analyzed.
- the objective of the experiment was to determine whether or not the malignancy itself was the source of T h2 cytokines that were detected in plasma.
- T h1 vs. T h2 cytokines IFN- ⁇ vs. IL-4, IL-5, IL-10, and IL-13, FIG. 6 b
- T h2 cytokine predominance in plasma was not derived from the tumor.
- the most highly/frequently up-regulated transcript in the tumor samples was VEGF ( FIG. 6 b ).
- Plasma VEGF levels were significantly higher in metastatic melanoma patients relative to healthy donors, ( FIG. 6 c ), consistent with published reports (Tas et al., Melanoma Res., 16:405-11 (2006)).
- VEGF immune modulatory (down-regulatory) properties of VEGF (Gabrilovich et al., Nat. Med., 2:1096-103 (1996))
- tumor derived VEGF could be responsible for the T h2 polarization in patients with stage IV melanoma (away from the normal state of T h1 dominance).
- VEGF has been associated with DC polarization towards DC 2 leading to Th 2 immune responses (e.g., asthma).
- Th 2 cytokines e.g., IL-4, IL-5, and IL-13
- IL-4, IL-5, and IL-13 have been associated with increased production of VEGF by a range of different cell types including smooth muscle cells.
- Preliminary data demonstrated that the addition of recombinant human VEGFA to a 2-day culture of normal blood-donor derived PBMC appeared to shift Th polarity away from Th 1 and towards Th 2 in a dose dependent fashion ( FIG. 7 ).
- VEGF vascular endothelial growth factor
- healthy donor PBMC were stimulated with CD3 + /CD28 + expander micro-beads for 3 days with increasing concentrations (1 pg/mL-16 pg/mL) of recombinant VEGF and assessed intracellular cytokine production of IL-4 (T h2 cytokine) and IFN- ⁇ (T h1 cytokine) in CD4 + T-cells at the end of in vitro culture ( FIG. 8 a ).
- T h2 cytokine IL-4
- IFN- ⁇ T h1 cytokine
- Increased concentrations of VEGF were associated with a decrease in the number of T h1 cells (CD4 + /IFN ⁇ + ) with an associated reciprocal increase in T h2 cells (CD4 + /IL-4 + ).
- the polarizing effects of VEGF were lost if the assay was performed with purified CD4 cells only ( FIG. 8 b ) suggesting that the observed T h polarization effect of VEGF is indirect, likely mediated by other PBMC.
- frozen peripheral blood specimens were randomly selected from a recently completed clinical trial (N047a) where patients with metastatic melanoma were treated with chemotherapy (paclitaxel+carboplatin) and a specific anti-VEGFA antibody (bevacizumab). They were analyzed for changes in plasma VEGFA levels and Th 1/ Th 2 polarity as well as frequency of tumor specific CTL (tetramer assay).
- the effect of chemotherapy may also have depleted (lymphodepleted) the pre-existing state of “chronic inflammation”; and the VEGF inhibitor (bevacizumab) may have allowed reconstitution of tumor-specific immunity in a Th 1 (not Th 2 ) dominant systemic environment.
- the additional, unanticipated, immunomodulatory effect of this therapy may have added to the observed therapeutic clinical result.
- Th 2 driven systemic chronic inflammation in patients with advanced melanoma (and possibly other malignancies) and reconstitution of effective immunity (Th 1 dominance) could potentially translate into effective therapy with clinically meaningful results. Therefore, an improved understanding of this mechanism of tumor mediated immune dysfunction/tumor progression as a function of Th 2 -mediated chronic inflammation could yield therapeutic targets for cancer therapy with agents already in clinical development for Th 2 mediated disorders (e.g., anti-IL-4 antibody).
- the identified cytokine profiles of plasma suggests the existence of a Th 2 dominant systemic immune environment in the blood of patients with metastatic melanoma.
- the analysis of the cellular/functional counterpart of the immune response in these patients across all stages of melanoma remains unknown.
- Th 2 dominant systemic immunity the existence of reciprocal, Th 2 polarized, changes in the cellular immune response in these patients across stages of melanoma that will correlate with the described changes in plasma cytokine and VEGF concentrations is determined.
- one can (a) enumerate Th 1 , Th 2 and T reg cells across stages of melanoma; (b); analyze the numbers and functional/differentiation state of circulating DC (DC 1 /DC 2 ) across stages of melanoma (DC 2 driven Th 2 polarization) and (c) analyze the functional status (active vs. tolerant) of both tumor-specific (e.g.
- EBV, CMV recall antigen specific
- Th 1 , Th 2 and T reg cells Enumeration of Th 1 , Th 2 and T reg cells across stages of melanoma.
- Th 2 cytokine predominance in plasma of patients with metastatic melanoma is truly a reflection of a systemic immune polarization towards Th 2 driven chronic inflammation
- the available frozen PBMC can be thawed and analyzed for the relative numbers of Th 1 , Th 2 and T reg .
- CD4 cells can be isolated from thawed PBMC specimens using paramagnetic beads coated with anti-CD4 (Dynal, Oslo, Norway), and they can be incubated with mouse-anti-human CD3/CD28 coated “stimulator” micro-beads (R and D Systems Minneapolis, Minn.) for 6 hours in the presence of 1 ug/ml brefeldin A, (Sigma Aldrich, St Louis, Mo.). After stimulation, the cells can be fixed, permeablized, and stained with APC conjugated mouse anti-human CD4 (Becton Dickinson, San Jose, Calif.) and FITC conjugated mouse anti-human IFN ⁇ and anti-IL-13 (R and D Systems Minneapolis, Minn.).
- APC conjugated mouse anti-human CD4 Becton Dickinson, San Jose, Calif.
- FITC conjugated mouse anti-human IFN ⁇ and anti-IL-13 R and D Systems Minneapolis, Minn.
- the stained samples can be analyzed by flow cytometry (FACScan and Cellquest software (Becton-Dickinson, San Jose, Calif.). The results can indicate the percentage of IFN ⁇ positive/IL-13 (or IL-4) negative (Th 1 ) and IFN ⁇ negative/IL-13 (or IL-4) positive (Th 2 ) helper T cells.
- Enumeration of T reg can be performed using intracellular staining for FoxP3 of CD4/25 positive lymphocytes
- Immunophenotyping can be conducted using commercially available monoclonal antibodies (Biolegend; San Diego, Calif.). Samples can be analyzed by flow-cytometry by FACScan® and data processed using Cellquest® software (Becton-Dickinson, Franklin Lakes, N.J.).
- the available, frozen PBMC corresponding to the plasma cytokine samples described above can be thawed and analyzed for the relative numbers of DC subsets defined by expression of CD11c + /CD123-(DC 1 ), and CD11c-/CD123 + (DC 2 ).
- Each subset can be analyzed for surface expression of co-stimulatory molecules (CD80, 83, 86)
- Immunophenotyping can be conducted using commercially available monoclonal antibodies (BD Pharmingen; San Jose, Calif.). Samples can be analyzed by flow-cytometry by FACScan® and data processed using Cellquest® software (Becton-Dickinson, Franklin Lakes, N.J.).
- the predominant phenotype of the tumor (and recall) antigen specific CTL can be one of tolerance (inability to synthesize intracellular IFN ⁇ upon congnant stimulation with tumor-specific peptides) and exhaustion (expression of PD-1). The latter has already been suggested to be true (Rosenberg et al., J.
- Immunophenotyping of PBMC can be performed using tetramers for melanoma differentiation antigen specific, HLA-A2 congnant peptides (MART-1 27-35 , gp100 209-217 and tyrosinase 368-376 ) as well as A2 cognant peptides of EBV and CMV (Beckman Coulter, San Diego, Calif.).
- PBMC thawed PBMC can be stained with FITC conjugated anti-CD8, PC5 conjugated anti-human CD4, CD14 and CD19, and conjugated HLA-A2 tetramers containing peptides from CMV, EBV (controls), MART-1 27-35 , gp100 209-217 and tyrosinase 368-376 .
- Samples can be analyzed by flow-cytometry and data processed using Cellquest® software (Becton-Dickinson, Franklin Lakes, N.J.). Gates can be set on lymphocytes that were CD4, CD14, and CD19 (PC5) negative and CD8 (APC) positive.
- tetramer positive CTL Functional analysis of tetramer positive CTL can be performed in patient samples demonstrating tetramer frequencies of at least 0.1% to melanoma differentiation or recall antigens.
- IFN ⁇ interferon-y
- aAPC artificial antigen presenting cell
- PBMC previously frozen patient PBMC can be thawed in batches, labeled with PE conjugated tetramers (Beckman Coulter, San Diego, Calif.), and stimulated for 6 hours, in the presence of 1 mg/mL brefeldin A, (Sigma Aldrich, St Louis, Mo.) with pararamagnetic beads (Dynal, Oslo, Norway) coated with peptide loaded HLA-A2 (Beckman Coulter San Diego, Calif.) and mouse anti-human CD28 (R and D Systems Minneapolis, Minn.).
- PE conjugated tetramers Beckman Coulter, San Diego, Calif.
- the cells After stimulation, the cells can be fixed, permeablized, and stained with APC conjugated mouse anti-human CD8 (Becton Dickinson, San Jose, Calif.) and FITC conjugated mouse anti-human IFN-gamma (R and D Systems Minneapolis, Minn.).
- the stained samples can be analyzed by flow cytometry (FACScan and Cellquest software (Becton-Dickinson, San Jose, Calif.).
- the results can indicate the percentage of tetramer positive CTL able vs. unable to synthesize intracellular IFN ⁇ .
- Ongoing QA data suggests an inter-assay variability with a CV of below 9%. Standard control samples can be run alongside all experiments. If the standard control samples generate results beyond ⁇ 2SD of the mean, all assay results can be rejected, and the experiment repeated.
- melanoma-specific therapeutic interventions on the VEGF/Th 2 state of tumor-induced chronic inflammation can be examined using peripheral blood biospecimens from patients with stage IV malignant melanoma enrolled on ongoing or completed clinical trials for changes in a range of immune parameters with a primary focus on Th 1 /Th 2 balance and functional tumor specific CTL immunity.
- the trials are listed in the Table 5.
- the patients enrolled into these trials had a blood specimen collected before initiation of therapy and after one cycle of treatment. This can provide data on the immediate impact of therapy on our VEGF/Th2/immune parameters of interest.
- the biospecimens collected before initiation of therapy and after one cycle of treatment can be used. This can provide data on the immediate impact of therapy on VEGF/Th2/immune parameters of interest. Additional testing for later time-points can be pursued only if justified by the initial analysis suggesting beneficial changes in the studied immune parameters. This can allow one to gain insight into the effects of a broad range of clinical interventions on immune homeostasis using an available (but limited) resource of biospecimens and only pursue further analysis if justified by the generated data.
- Laboratory analyses of the stored PBMC can include the same assays described above and can also include: (a) PBMC immunophenotyping for immune cell subset analysis; and (b) plasma cytokine profiling.
- PBMC immunophenotyping for immune cell subset analysis.
- Pre and post-treatment frozen PBMC biospecimens can be analyzed for the “global” impact of therapy on immune cell subsets.
- the stained samples can be analyzed by flow cytometry (FACScan and Cellquest software (Becton-Dickinson, San Jose, Calif.).
- PBMC isolated from patients prior to B7-DC XAb and 15 days after antibody treatment can be assayed. Changes in the numbers of cells bearing the lymphocyte markers in pared comparisons for patients prior to B7-DC XAb treatment can be used to ascertain antibody treatment effects.
- Plasma cytokine profiling In order to complement the PBMC derived cellular immunity analyses, one can add plasma cytokine measurements in the same samples (paired PBMC and plasma testing). To that end, one can profile the serum cytokine changes as a result of specific therapy for all available specimens before and after treatment.
- the BioRad human 27-plex cytokine panel can be used (Cat # 171-A11127, Bio-Rad, San Diego Calif.) for the measurements of plasma concentrations of IL-1 ⁇ , IL-1r ⁇ , IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12 (p70), IL-13, IL-15, IL-17, basic FGF, Eotaxin, G-CSF, GM-CSF, IFN- ⁇ , IP-10, MCP-1, MIP-1 ⁇ , MIP-1 ⁇ , PDGF, RANTES, TNF- ⁇ , and VEGF.
- the assay can be performed as per the manufacturer's directions.
- Bio-Plex assay buffer 100 ⁇ L of Bio-Plex assay buffer can be added to each well of a MultiScreen MABVN 1.2 ⁇ m microfiltration plate followed by the addition of 50 ⁇ L of the multiplex bead preparation. Following washing of the beads with the addition of 100 ⁇ L of wash buffer, 50 ⁇ L the samples or the standards can be added to each well and incubated with shaking for 30 minutes at room temperature.
- the plasma (1:3 dilution) and standards can be diluted using the Bio-Plex human serum diluent kit and plated in duplicate. Standard curves can be generated with a mixture of 27 cytokine standards and eight serial dilutions ranging from 0-32,000 pg/mL.
- the plate can then be washed 3 times followed by incubation of each well in 25 ⁇ L of pre-mixed detection antibodies for 30 minutes with shaking.
- the plate can further be washed and 50 ⁇ L of streptavidin solution were added to each well and incubated for 10 minutes at room temperature with shaking.
- the beads can be given a final washing and resuspension in 125 ⁇ L of Bio-Plex assay buffer.
- Cytokine levels in the sera can be quantified by analyzing 100 ⁇ L of each well on a Bio-Plex using Bio-Plex Manager software version 4.0. Normal values for plasma cytokine concentrations were generated by analyzing 30 plasma samples from healthy donors (blood donors at the Mayo Clinic Dept. of Transfusion Medicine). A set of five normal plasma samples (standards) can be run along side all batches of plasma analysis. If the cytokine concentrations of the “standard” samples differ by more than 20%, results can be rejected, and the plasma samples re-analyzed.
- the clinical trail can be conducted in the context of the clinical trials program of the Melanoma Study Group of the Mayo Clinic Cancer Center. All patients with the diagnosis of metastatic melanoma that are planned to undergo complete surgical resection of their malignancy can be offered participation in this study.
- the objective of the study can be to profile the changes in immune homeostasis from pre-surgery, post-surgery and all through the time of clinically detectable tumor relapse. It is hypothesized that the state of VEGF/Th 2 driven chronic inflammation can be established pre-surgery, resolved soon after surgery and slowly re-develop in the months prior to clinical tumor relapse.
- patients can be clinically followed in accordance to clinical practice (every 2 months). Patients can be asked to donate 100 mL of blood at each follow-up time point. The blood can be collected, processed, and stored in accordance to existing procedures for immunological testing Immune homeostasis analysis can be conducted in batches to limit inter-assay variability. Specific focus/priority can be given to parameters reflecting Th 1 /Th 2 balance, frequency of functional/tolerant tumor (or recall) antigen specific CTL as well as plasma cytokine and VEGF levels. At the time of clinical relapse, patients can be re-tested, and the tumor biopsied for histologic confirmation.
- tumor associated antigens immunohistochemistry for MART-1, gp100 and tyrosinase
- tumor infiltrating lymphocytes are available tumor tissues.
- tumor associated antigens immunohistochemistry for MART-1, gp100 and tyrosinase
- tumor infiltrating lymphocytes are available tumor tissues.
- patients can undergo another curative surgical resection at the time of relapse, they may continue on study following the outlined follow-up/testing schedule until such a time when a tumor relapse is no longer surgically resectable. See, e.g., Table 6.
- CD4 T cells (Th1 and Th2 CD4 T cells) derived from human PBMC were incubated in vitro with varying concentrations of VEGFA (rhVEGFA; 10 ng/mL, 50 ng/mL, and 200 ng/mL) or TGF ⁇ (rhTGF ⁇ ; 10 ng/mL, 50 ng/mL, and 200 ng/mL). After six hours of incubation at 37° C., the ratio of Th1 vs. Th2 (Th1/Th2) was determined Both VEGFA and TGF ⁇ exhibited a similar effect on Th1/Th2 polarity in human PBMC derived CD4 cells ( FIG. 12 ).
- VEGFA rhVEGFA
- TGF ⁇ TGF ⁇
- CD4 T cells (Th1 and Th2 CD4 T cells) derived from human PBMC were incubated in vitro with VEGFA alone (1 ng/mL, 5 ng/mL, 10 ng/mL, 100 ng/mL, or 1000 ng/mL) or VEGFA (100 ng/mL) plus an anti-TGF ⁇ antibody (1 ng/mL, 10 ng/mL, 100 ng/mL, 1 ⁇ g/mL, 5 ⁇ g/mL, or 10 ⁇ g/mL).
- the anti-TGF ⁇ antibody was obtained from Genzyme Corp. (Cambridge, Mass.). Untreated cells (media only) as well as cells exposed to Th1 or Th2 favorable in vitro conditions were used as controls.
- Th1/Th2 Th1/Th2
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Oncology (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biochemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Endocrinology (AREA)
- Cell Biology (AREA)
- Pain & Pain Management (AREA)
- Rheumatology (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
This document relates to methods and materials involved in treating cancer (e.g., melanoma). For example, methods and materials involved in using an anti-chronic inflammation treatment (e.g., chemotherapy) in combination with a cancer treatment agent (e.g., a cancer vaccine) to treat cancer are provided.
Description
- This application is a continuation-in-part of International Application No. PCT/US2009/049511, filed Jul. 2, 2009, which claims the benefit of priority from U.S. Provisional Application Ser. No. 61/078,203, filed on Jul. 3, 2008. The disclosures of the prior applications are considered part of (and are incorporated by reference in) the disclosure of this application.
- 1. Technical Field
- This document relates to methods and materials involved in treating cancer (e.g., melanoma). For example, this document relates to methods and materials involved in using an anti-chronic inflammation treatment (e.g., chemotherapy) in combination with a cancer treatment agent (e.g., a cancer vaccine) to treat cancer.
- 2. Background Information
- Cancer is a serious illness that affects many people every year. Melanoma is the most serious form of skin cancer. It is a malignant tumor that originates in melanocytes, the cells which produce the pigment melanin that colors skin, hair, and eyes and is heavily concentrated in most moles. While it is not the most common type of skin cancer, melanoma underlies the majority of skin cancer-related deaths. About 48,000 deaths worldwide are registered annually as being due to malignant melanoma. Worldwide, there are about 160,000 new cases of melanoma each year. Melanoma is more frequent in white men and is particularly common in white populations living in sunny climates. Other risk factors for developing melanoma include a history of sunburn, excessive sun exposure, living in a sunny climate or at high altitude, having many moles or large moles, and a family or personal history of skin cancer.
- This document provides methods and materials related to treating cancer. For example, this document provides methods and materials for using an anti-chronic inflammation treatment (e.g., chemotherapy) in combination with a cancer treatment agent (e.g., a cancer vaccine) to treat cancer. As described herein, cancer can induce a global state of immune dysfunction and/or chronic inflammation in the cancer patient. This global state of immune dysfunction and/or chronic inflammation can prevent the patient from mounting a successful response against the cancer. For example, a cancer patient with a global state of chronic inflammation can be in a state such that the patient is unable to generate an anti-cancer immune response when given an anti-cancer vaccine. The methods and materials provided herein can be used to reduce the global state of immune dysfunction and/or chronic inflammation present within a cancer patient such that the cancer patient can better respond to a cancer treatment such as a cancer vaccine. As described herein, chemotherapy, radiation, anti-IL-4 agents (e.g., anti-IL-4 antibodies), anti-IL-13 agents (e.g., soluble IL-13 receptor), steroids, and combinations thereof can be used to reduce the global state of immune dysfunction and/or chronic inflammation present within a cancer patient. Once the global state of immune dysfunction and/or chronic inflammation present within a cancer patient is reduced, the cancer patient can be treated with an appropriate cancer treatment such as a cancer vaccine or other immune stimulating agents (e.g., IL-2 or IL-12).
- In general, this document features a method for treating a mammal having cancer. The method comprises, or consists essentially of, (a) administering to the mammal an anti-chronic inflammation treatment under conditions wherein the level of global chronic inflammation in the mammal is reduced, and (b) administering to the mammal a cancer treatment agent under conditions wherein the presence of the cancer is reduced. The mammal can be a human. The cancer can be melanoma. The cancer can be stage IV melanoma. The anti-chronic inflammation treatment can comprise chemotherapy, radiation, an anti-IL-4 agent, an anti-IL-13 agent, or a steroid treatment. The cancer treatment agent can be a cancer vaccine. The cancer vaccine can be a MART-1, gp100, or survivin cancer vaccine. The period of time between the last administration of the anti-chronic inflammation treatment and the first administration of the cancer treatment agent can be between two weeks and six months.
- In another aspect, this document features a method for treating a mammal having cancer. The method comprises, or consists essentially of, (a) administering to the mammal an anti-TGFβ antibody under conditions wherein the level of global chronic inflammation in the mammal is reduced, and (b) administering to the mammal a cancer treatment agent under conditions wherein the presence of the cancer is reduced. The mammal can be a human. The cancer can be melanoma. The cancer can be stage IV melanoma. The cancer treatment agent can be a cancer vaccine. The cancer vaccine can be a MART-1, gp100, or survivin cancer vaccine. The period of time between the last administration of the anti-TGFβ antibody and the first administration of the cancer treatment agent can be between two weeks and six months.
- Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although methods and materials similar or equivalent to those described herein can be used to practice the invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.
- The details of one or more embodiments of the invention are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims.
-
FIG. 1 : Effects of 1% patient plasma (vs. normal plasma) on in vitro maturation of normal DC. Presented is a representative experiment demonstrating the % co-stimulatory molecule positive (CD80, 83, 86) DC. -
FIG. 2 . Allogeneic mixed lymphocyte reaction cultures evaluating proliferation of T cells mixed at differing ratios with mature or immature dendritic cells in the presence of normal (control) plasma or plasma from a patient with metastatic melanoma (MTB plasma). T cell proliferation was assessed by 3H-TdR incorporation. Similar results were observed in two other experiments. -
FIG. 3 . Three dimensional representation of the results of Principal Component Analysis (PCA). The PCA was performed on 234 clinical samples for concentrations of 27 cytokines. Each sphere represents one clinical sample. The dark spheres represent stage IV melanoma patients, and the lighter spheres represent atypical nevi, benign nevi, in situ melanoma, stage I melanoma, stage II melanoma, or stage III melanoma. The axes are the main principal components. Significant grouping is only evident for patients in the cohort of stage IV (metastatic) melanoma (dark spheres). -
FIG. 4 : Mean plasma IL-4 levels (pg/mL)±SD across stages of melanoma (melanoma in situ and stages: I, II, III, IV), patients with atypical nevi (atypical) and benign nevi (normal controls). -
FIG. 5 : Assessment of T-cell phenotype and function in healthy donors and stage IV melanoma patients. The number of T-cells exhibiting the FoxP3 (Treg) or PD-1 phenotype in peripheral blood was determined in healthy donors and stage IV melanoma patients (A). The frequency of FoxP3 positive cells were measured by 3-color flow cytometry, CD4-PC5, CD25-PE and FoxP3-Alexa flour 488. The mean percent (+/−SD) of FoxP3 positive were determined from the CD4 and CD25 double positive population. The mean frequency (+/−SD) of PD-1+ cells was measured from the CD8+ population. The frequency (+/−SD) of tetramer positive (CMV or MART-1) CD8+ T cells was compared among normal volunteers and patients with stage IV melanoma (B). -
FIG. 6 : VEGF levels in patients with metastatic melanoma. (A) RNA expression of cytokines in human metastatic melanoma tissue. Twenty-four frozen biopsies of metastatic melanoma tumor tissues was used to extract RNA for expression array analysis. Illustrated are the RNA expression intensity profiles of 45 probes for 24 cytokines. (B) Comparison of expression intensities between genes coding for Th1 (IFN-γ and IL-2), Th2 (IL-4, IL-5, IL-10, and IL-13) cytokines and VEGF. There were no statistically significant differences when comparing Th1 vs. Th2 cytokine expression levels (p=0.04). There was a statistically significant difference when comparing VEGF expression with Th1 or Th2 cytokines (p<0.001). Levels of significance were determined using the Wilcoxin signed-rank test. (C) ELISA (mean concentration +/−SD) for VEGF-A was performed on plasma samples from healthy donors (n=30) and stage IV melanoma patients (n=40). -
FIG. 7 . Healthy donor PBMCs were cultured in vitro for 48 hours in the presence of increasing concentrations of recombinant human VEGFA. At the end of the incubation, cells were stimulated with PMA and ionomycin, in the presence of brefeldin A and stained for intracellular IFNγ, IL-4, or IL-13 and surface immunophenotyped for CD294 or TIM-3. Cells were then analyzed for the frequency of CD4 cells (% of CD4) expressing said phenotypes using flow cytometry. -
FIG. 8 : Co-culture with recombinant human VEGF shifts T-helper polarity from Th1 (IFN-γ) to Th2 (IL-4) predominance. PBMC (A) isolated from healthy donors were stimulated with PMA and ionomycin in the presence of brefeldin-A, permeabolized, and intracellularly stained for human IFN-γ (FITC) and human IL-4 (PE). PBMC were exposed to increasing concentrations of VEGF (0-16 pg/mL) without/with IL-12. All cells were immunostained with PC5 anti-human CD4. Purified CD4+ T-cells (B) were negatively isolated using Miltenyi beads, cultured, and stained in the same fashion as PBMC (A). Similar results were observed in 5 different experiments. -
FIG. 9 . Changes in plasma VEGF levels (plasma VEGFA in pg/mL; top left) at three time points in a single patient with metastatic melanoma treated on protocol N047a correlate with improved Th1/Th2 ratio as determined by intracellular staining of CD4+ cells for IFN gamma or IL-4 (top right). These also correlate with emergence of increased frequencies of tumor specific CTL (bottom right) as determined by tetramer assay. -
FIG. 10 . Cellular interactions of acute and chronic inflammation. MSC (myeloid suppressor cell); Th1 & Th2 (T helper lymphocytes type 1 & 2); Treg: regulatory T cell; DC1 (dendritic cells, type 1); DC2 (dendritic cells type 2); CTL (cytotoxic T lymphocyte); illustrated is central role of tumor derived VEGF in polarizing immunity towards Th2 mediated “chronic inflammation”. -
FIG. 11 : Correlation of surface immunophenotyping for CD294 and TIM-3 with intracellular IL-4, IL-13 and IFNγ for the purposes of enumeration of Th2 and Th1 cells respectively. -
FIG. 12 : Changes in the ratio of human PBMC derived CD4 T cell subsets (Th1 vs. Th2) following in vitro incubation with varying concentrations of VEGFA or TGFβ. These results indicate that both VEGFA and TGFβ have a similar effect on Th1/Th2 polarity in human PBMC derived CD4 cells. -
FIG. 13 : Relative ratios of human PBMC derived CD4 T cells subsets (Th1 vs. Th2) cultured in vitro with varying concentrations of VEGFA in the absence or presence of increasing concentrations of anti-TGFβ neutralizing antibody. Untreated (media) as well as Th1 (Th1) and Th2 (Th2) favorable in vitro conditions are presented as controls. These results indicate that presence of anti-TGFβ antibodies reverses the Th1/Th2 modulation of VEGFA in vitro, suggesting that the observed VEGF effect in these cells may be TGFβ mediated. - This document provides methods and materials related to treating cancer in mammals. For example, this document provides methods and materials related to the use of an anti-chronic inflammation treatment (e.g., chemotherapy) in combination with a cancer treatment agent (e.g., a cancer vaccine) to treat cancer.
- The methods and materials provided herein can be used to treat cancer in any type of mammal including, without limitation, mice, rats, dogs, cats, horses, cows, pigs, monkeys, and humans. Any type of cancer, such as skin cancer (e.g., melanoma), can be treated. Examples of cancer that can be treated as described herein include, without limitation, skin cancer, lung cancer, breast cancer, prostate cancer, ovarian cancer, and colon cancer. In some cases, stage I, stage II, stage III, or stage IV melanoma can be treated using the methods and materials provided herein.
- In general, cancer can be treated by administering an anti-chronic inflammation treatment such that the global state of immune dysfunction and/or chronic inflammation present within a cancer patient is reduced. For example, chemotherapy, radiation, anti-IL-4 agents (e.g., anti-IL-4 antibodies), anti-IL-13 agents (e.g., soluble IL-13 receptor), steroids, and combinations thereof can be used to reduce the global state of immune dysfunction and/or chronic inflammation present within a cancer patient. In some cases, chemotherapy such as paclitaxel, carboplatin, temozolomide, or cyclophosphamide can be administered to a cancer patient to reduce the global state of immune dysfunction and/or chronic inflammation present within a cancer patient. In some cases, an anti-chronic inflammation treatment can include, without limitation, administering an anti-TGFβ antibody. For example, anti-TGFβ antibodies can be administered to a cancer patient to reduce the global state of immune dysfunction and/or chronic inflammation present within a cancer patient. Examples of anti-TGFβ antibodies include, without limitation, human monoclonal anti-TGF-β1 antibodies such as CAT-192 (Genzyme Inc.).
- Any appropriate method can be used to assess whether or not the global state of immune dysfunction and/or chronic inflammation was reduced following an anti-chronic inflammation treatment. For example, cytokine profiles (e.g., IL-4, IL-13, IL-4, IL-13, IL-5, IL-10, IL-2, and interferon gamma) present in blood can be assessed before and after an anti-chronic inflammation treatment to determine whether or not the global state of immune dysfunction and/or chronic inflammation was reduced.
- Once the global state of immune dysfunction and/or chronic inflammation present within a cancer patient is reduced, the cancer patient can be treated with an appropriate cancer treatment (e.g., an immune cancer treatment) such as a cancer vaccine. Examples of appropriate cancer treatment agents include, without limitation, immune stimulating cytokines (e.g., IL-2, IL-12, interferon alpha, and interferon gamma), inhibitors of immune down-regulation (e.g., anti-CTLA4, anti-41bb, anti-PD-1, and anti-CD25), and cancer vaccines (e.g., MART-1, gp100, survivin, and tyrosinase cancer vaccines). It will be appreciated that paclitaxel, carboplatin, bevacizumab, and anti-CTLA-4 can be used to treat (e.g., skin cancer) upon administration either individually or in any combination thereof (e.g., paclitaxel, carboplatin and bevacizumab).
- In some cases, the amount of time between administration of an anti-chronic inflammation treatment and administration of a cancer treatment can be between two weeks and twelve months (e.g., between two weeks and eleven months, between two weeks and ten months, between two weeks and nine months, between two weeks and eight months, between two weeks and seven months, between two weeks and six months, between one month and twelve months, between one month and six months, or between two months and six months). For example, a chemotherapy agent (e.g., paclitaxel) can be administered to a cancer patient to reduce the global state of immune dysfunction and/or chronic inflammation present within the cancer patient. Then, after one month without any type of anti-chronic inflammation treatment, a cancer treatment (e.g., a cancer vaccine) can be administered to the cancer patient.
- Any appropriate method can be used to administer a cancer treatment agent to a mammal For example, a cancer treatment agent can be administered orally or via injection (e.g., subcutaneous injection, intramuscular injection, intravenous injection, or intrathecal injection). In some cases, cancer treatment agents can be administered by different routes. For example, one cancer treatment agent can be administered orally and a second cancer treatment agent can be administered via injection.
- In some cases, a cancer treatment agent can be administered following resection of a tumor. Cancer treatment agent can be administered to a mammal in any amount, at any frequency, and for any duration effective to achieve a desired outcome (e.g., to increase progression-free survival or to increase the time to progression). In some cases, cancer treatment agents can be administered to a mammal having skin cancer to reduce the progression rate of melanoma by 5, 10, 25, 50, 75, 100, or more percent. For example, the progression rate can be reduced such that no additional cancer progression is detected. Any method can be used to determine whether or not the progression rate of skin cancer is reduced. For example, the progression rate of skin cancer can be assessed by imaging tissue at different time points and determining the amount of cancer cells present. The amounts of cancer cells determined within tissue at different times can be compared to determine the progression rate. After treatment as described herein, the progression rate can be determined again over another time interval. In some cases, the stage of skin cancer after treatment can be determined and compared to the stage before treatment to determine whether or not the progression rate was reduced.
- In some cases, a cancer treatment agent can be administered to a mammal having cancer under conditions where progression-free survival or time to progression is increased (e.g., by 5, 10, 25, 50, 75, 100, or more percent) as compared to the median progression-free survival or time to progression, respectively, of corresponding mammals having untreated cancer.
- An effective amount of a cancer treatment agent can be any amount that reduces the progression rate of cancer, increases the progression-free survival rate, or increases the median time to progression without producing significant toxicity to the mammal Typically, an effective amount of a cancer treatment agent such as bevacizumab can be from about 5 mg/kg/week to about 15 mg/kg/week (e.g., about 10 mg/kg/week). If a particular mammal fails to respond to a particular amount, then the amount of one or more of the compounds can be increased by, for example, two fold. After receiving this higher concentration, the mammal can be monitored for both responsiveness to the treatment and toxicity symptoms, and adjustments made accordingly. The effective amount can remain constant or can be adjusted as a sliding scale or variable dose depending on the mammal's response to treatment. Various factors can influence the actual effective amount used for a particular application. For example, the frequency of administration, duration of treatment, use of multiple treatment agents, route of administration, and severity of the cancer may require an increase or decrease in the actual effective amount administered.
- The frequency of administration can be any frequency that reduces the progression rate of cancer, increases the progression-free survival rate, or increases the median time to progression without producing significant toxicity to the mammal For example, the frequency of administration can be from about once a month to about three times a month, or from about twice a month to about six times a month, or from about once every two months to about three times every two months. The frequency of administration can remain constant or can be variable during the duration of treatment. In addition, the frequency of administration of multiple cancer treatment agents can be the same or can differ. For example, one cancer treatment agent can be administered three times during a 28 day period, while a second cancer treatment agent can be administered one time, and third cancer treatment agent can be administered two times during the same period. A course of treatment with a cancer treatment agent can include rest periods. For example, a cancer treatment agent can be administered over a two week period followed by a two week rest period, and such a regimen can be repeated multiple times. As with the effective amount, various factors can influence the actual frequency of administration used for a particular application. For example, the effective amount, duration of treatment, use of multiple treatment agents, route of administration, and severity of the cancer may require an increase or decrease in administration frequency.
- An effective duration for administering a composition provided herein can be any duration that reduces the progression rate of cancer, increases the progression-free survival rate, or increases the median time to progression without producing significant toxicity to the mammal Thus, the effective duration can vary from several days to several weeks, months, or years. In general, the effective duration for the treatment of cancer can range in duration from several weeks to several months. In some cases, an effective duration can be for as long as an individual mammal is alive. Multiple factors can influence the actual effective duration used for a particular treatment. For example, an effective duration can vary with the frequency of administration, effective amount, use of multiple treatment agents, route of administration, and severity of the cancer.
- After administering a composition provided herein to a mammal, the mammal can be monitored to determine whether or not the cancer was treated. For example, a mammal can be assessed after treatment to determine whether or not the progression rate of cancer was reduced (e.g., stopped). As described herein, any appropriate method can be used to assess progression and survival rates.
- The invention will be further described in the following examples, which do not limit the scope of the invention described in the claims.
- Blood samples collected from patients with early stage melanoma (melanoma in situ and melanoma stage I, II and III) and benign nevi (atypical/dysplastic nevi) were newly diagnosed patients with no previous treatment. All patients were tumor free at the time of peripheral blood collection. Samples from patients with metastatic melanoma (newly diagnosed, previously untreated) as well as healthy volunteers/controls were collected under a separate melanoma blood and tissue banking protocol. Both protocols were reviewed and approved for use in these studies. All biospecimens were collected, processed, and stored in uniform fashion following established standard operating procedures. All patients signed an informed consent document. The presented study describes data obtained from 113 men and 96 women ranging in age from 21 to 85 (Table 1).
-
TABLE 1 Study patient population distributed by clinical category, age, sex and assayed immune parameters. Assayed immune parameters Age T-cell Total mean ± SD Cell Plasma Function RNA Clinical category Patients (range) % female Subset Cytokines Tetramer Assay array Benign Nevi 34 51 ± 12 68 26 34 7 2 0 (21-71) Atypical/ Dysplastic 25 52 ± 16 44 22 16 11 1 0 (25-84) In situ melanoma 36 61 ± 16 36 30 35 16 3 0 (26-84) Stage I 45 54 ± 17 44 36 44 16 4 0 (21-82) Stage II 16 55 ± 17 44 11 12 9 0 0 (22-81) Stage III 16 53 ± 19 44 14 16 6 1 0 (23-83) Stage IV 37 56 ± 14 43 32 30 27 16 24 (28-85) - Peripheral venous blood (50-90 mL) was drawn into heparinized Vacutainer tubes that were processed and separated into plasma and peripheral blood mononuclear cells (PBMC) following gradient centrifugation using Ficoll-Paque (GE Healthcare Uppsala, Sweden). Plasma was collected and immediately frozen at −80° C. (1 mL aliquots). PBMC were collected, washed in phosphate buffered saline (PBS), counted, diluted to 1×107/mL and viably frozen in 90% cosmic calf serum (Hyclone Inc. Logan, Utah) and 10% DMSO (Sigma St. Louis, Mo.). All assays were batch-analyzed at the end of the study.
- The following anti-human monoclonal antibodies were used in PBMC immunophenotyping for flow cytometry: anti-CD3-APC, FITC and PE, anti-CD4-FITC, anti-CD8-PE, anti-CD16 PE, anti-CD56 PE, anti-CD62L APC, anti-CD69 FITC, anti-CD14 FITC, anti-CD16 FITC, anti-CD19 FITC, anti-CD11c APC, anti-CD80 PE, anti-CD83 PE, anti-CD86 PE, anti-CD40 APC, anti-HLA-DR PC5, anti-PD-1 (BD Pharmingen San Jose, Calif.). The human monoclonal antibodies anti-CD4 PC5 and anti-CD25 PE were purchased from Biolegend (San Diego, Calif.) and used in conjunction with anti-human FoxP3 for the enumeration of Treg cells. The following anti-human monoclonal antibodies were used for intracellular staining for flow cytometry: anti-IFNβ FITC, anti-IL-13 PE, anti-IL-4 PE (R and D Systems Minneapolis, Minn.), and anti-FoxP3 Alexaflour 488 (Biolegend San Diego, Calif.).
- Previously frozen PBMC (0.5-1.0 ×106 cells/mL) were thawed and aliquoted into 96 well rounded bottom plates (100 μL/well). The desired antibody or antibody pool was added at 5 μL/well. The cells and antibodies were incubated for 30 minutes at 4° C. and washed twice with 1× PBS (Cellgro Manassas, Va.), 0.1% BSA and 0.05% sodium azide (Sigma St. Louis, Mo.). Four-color flow cytometry was performed on a LSRII flow cytometer (Becton Dickenson San Jose, Calif.), and Cellquest software (Becton Dickenson San Jose, Calif.) was utilized for data analysis. For dendritic cells, a gate was set on cells, which were HLA-DR+ and Lin− (CD3, CD14, CD16 and CD19). From this population the percentage of cells, which were CD11c+ and positive for costimulatory molecules (CD80, CD83 and CD86) was determined as previously elsewhere (Fricke et al., Clin. Cancer Res., 13:4840-8 (2007)). A panel of tumor associated antigen tetramers, MART-126-35, gp100264-272, gp100209-217, and tyrosinase369-377 (Beckman Coulter San Jose, Calif.) were used to enumerate the frequency of tumor antigen specific CD8 positive T-cells. Recall antigens, EBV280-288 and CMV495-503 (Beckman Coulter San Jose, Calif.) were used as positive controls. For tetramer frequencies, a gate was set on lymphocytes, which were CD8+ and negative for CD4, CD14 and CD 19. Three-color flow cytometry was performed on a LSRII flow cytometer (Becton Dickenson San Jose, Calif.) and Cellquest software (Becton Dickenson San Jose, Calif.) was utilized for data analysis.
- Functional enumeration of tumor antigen specific CTL was performed using an artificial antigen presenting cell method (aAPC) as described elsewhere (Markovic et al., Clin. Exp. Immunol., 145:438-47 (2006)). Briefly, frozen PBMC were thawed, labeled with the desired tumor antigen peptide/class I tetramers (Beckman Coulter Fullerton, Calif.) and stimulated for 6 hours with streptavidin coated microbeads (Invitrogen Oslo, Norway) loaded with HLA-A2 class I containing tumor antigen peptides of choice (MART-1, gp100 or tyrosinase) and anti-human CD28 in the presence of brefeldin A (Sigma, St. Louis, Mo.). After stimulation, the cells were fixed with 2% paraformaldehyde (Sigma, St. Louis, Mo.) and then permeabilized with 0.1% saponin (Sigma, St. Louis, Mo.) in PBS. Cells were then immunophenotyped with anti-human CD4-PC5 or CD8-APC and intracellular staining was done with anti-human IFNγ FITC or IL-4 PE. Four-color flow cytometry was performed with a FACSCaliber and Cellquest software (Becton Dickenson San Jose, Calif.) was utilized for data analysis.
- Protein levels for 27 cytokines, chemokines, and growth factors, including IL-1β, IL-1ra, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12p70, IL-13, IL-15, IL-17, Eotaxin, FGF basic, G-CSF, GM-CSF, IFN-γ, IP-10, MCP-1, MIP-1α, MIP-1β, PDGF, RANTES, TNF-α, and VEGF, were measured using the Bio-plex cytokine assay (Bio-rad, Hercules, Calif.) as per manufacturer's instructions. Patient plasma was diluted 1:4 in dilution buffer and 50 μL was added to washed, fluorescently dyed microspheres (beads) to which biomolecules of interest are bound. The beads and diluted patient plasma were incubated for 30 minutes at room temperature with agitation. After the incubation the beads were washed in Bio-plex wash buffer and placed in 25 μL of detection antibody and incubated for 30 minutes as described above. After washing, the beads were placed in streptavidin-PE, incubated, and washed a final time. The bound beads were resuspended in 125 μL Bio-plex assay buffer and read with the Luminex plate reader (Bio-rad, Hercules, Calif.). Protein concentrations were determined using a standard curve generated using the high PMT concentrations with sensitivity from 10-1000 pg/mL.
- To determine the effect of VEGF on Th1 and Th2 polarity, PBMC from healthy donors were stimulated for 3 days with CD3/CD28 expander beads (Invitrogen Oslo, Norway) with and without increasing doses of recombinant VEGF (1-16 pg/mL). Cells were also cultured with 10 μg/mL recombinant human IL-12 (R and D Systems, Minneapolis, Minn.) or 8 μg/mL of a monoclonal anti-human IL-12 (R and D Systems Minneapolis, Minn. clone #24910). After the culture, the cells were harvested and restimulated with 50 ng/mL PMA (Sigma, St. Louis, Mo.) and 1 μg/mL ionomycin (Sigma, St. Louis, Mo.) in the presence of 10 μg/mL brefeldin A for 4 hours. The cells were then stained with anti-human CD4, anti-human IFN-γ and anti-human IL-4 flow cytometry.
- Frozen tissue sections of melanoma biopsies, were examined, regions of pure tumor with little/no evidence of necrosis or stromal infiltration were outlined, scraped off the slides, and used for RNA extraction. Total RNA was isolated from the excised tumor tissue using the Qiagen RNA extraction kit (Qiagen Valencia, Calif.). The quality of the RNA was evaluated by obtaining electropherograms on Agilent 2100 Bioanalyzer and RNA integrity number (RIN) using 2100 Expert software (Agilent Technologies, Inc. Palo Alto, Calif.). cDNA was prepared from a total of 10 μg of RNA. Samples were quantified using standard spectrophotometry using a Tecan spectrophotometer (Tecan US, Research Triangle Park, N.C.) and considered acceptable if the A260/280 reading was >1.7. The purified cDNA was used as a template for in vitro transcription reaction for the synthesis of biotinylated cRNA using RNA transcript labeling reagent (Affymetrix, Santa Clara, Calif.). Labeled cRNA was then fragmented and hybridized onto the U133 Plus 2.0 array. Appropriate amounts of fragmented cRNA and control oligonucleotide B2 were added along with control cRNA (BioB, BioC, and BioD), herring sperm DNA, and bovine serum albumin to the hybridization buffer. The hybridization mixture was heated at 99° C. for 5 minutes followed by incubation at 45° C. for 5 minutes before injecting the sample into the microarray. Then, the hybridization was carried out at 45° C. for 16 hours with mixing on a rotisserie at 60 rpm. After hybridization, the solutions were removed, and the arrays were washed and then stained with streptavidin-phycoerythrin (Molecular Probes, Eugene, Oreg.). After washes, arrays were scanned using the GeneChip Scanner 3000 (Affymetrix, Santa Clara, Calif.). The quality of the fragmented biotin labeled cRNA in each experiment was evaluated before hybridizing onto the U133A expression array by both obtaining electropherograms on Agilent 2100 Bioanalyzer and hybridizing a fraction of the sample onto test-3 array as a measure of quality control. GeneSpring GX 7.3 (Agilent Technologies, Inc. Santa Clara, Calif.) data analysis software was used to analyze the results of the microarray experiment. Gene expression values were normalized by the GCRMA algorithm (Bolstad et al., Bioinformatics, 19:185-93 (2003)).
- The majority of samples analyzed in this report were randomly assigned to batches for each laboratory assay due to the fact that all samples were not collected/processed at the same time. The randomization was stratified to assure an even distribution across the stages of disease for each batch. The distributions of the results of each run were examined, and those that did not appear to be normally distributed were transformed using either logarithmic or square root transformations. In order to look at differences in various parameters between stages of disease, analysis was performed utilizing analysis of covariance (ANCOVA), adjusting for age, gender, and batch effects. Results of this analysis were summarized by least square means and 95% confidence intervals for each stage of disease. The p-values presented are those from the overall ANCOVA, which compares the means levels of each parameter across all stages of disease. P-values <0.05 were considered to be statistically significant. Due to the magnitude of the cytokine data from the multiplex assay the data was processed using Partek 6.3 software (Partek Inc. St. Louis Mo.) and analyzed using a principal component analysis (PCA) approach. PCA was utilized in an effort to vector space transform a multidimensional data set representing 27 variables for each individual patient and group patients based on similar cytokine concentrations revealing the internal relationships of cytokines within patient groups (e.g., per stage of melanoma) in an unbiased way.
- Preliminary results support the notion that systemic immune dysfunction can lead to the observed induction of tolerance following peptide vaccination in clinical trials. In this example, normal donor myeloid DC were exposed to in vitro culture conditions (GM-CSF, IL-4, and CD40L) that lead to their differentiation and maturation (expression of co-stimulatory molecules) (
FIG. 1 ). The addition of patient plasma to these experimental conditions resulted in a significant reduction in the number of DC expressing key co-stimulatory molecules (maturation), suggesting the presence of a soluble inhibitor(s) of DC maturation. Similar observations, with varying degrees of “suppression” were made in another nine experiments (nine other samples of patient plasma). Even when normal DCs (mature or immature) were combined with other normal donor lymphocytes in a mixed lymphocyte culture, the presence of patient vs. normal plasma lead to significantly diminished lymphocyte proliferation (FIG. 2 ). Thus, factors in the plasma of patients with metastatic melanoma are interrupting normal immune cell function. - The identity of the unknown factor(s) could be a known cytokine. Thus, a screening study was performed to quantify the plasma concentrations of 27 different cytokines (BioRad human 27-plex cytokine panel assaying for plasma concentrations of IL-1β, IL-1rα, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12 (p70), IL-13, IL-15, IL-17, basic FGF, eotaxin, G-CSF, GM-CSF, IFN-γ, IP-10, MCP-1, MIP-1α, MIP-1β, PDGF, RANTES, TNF-α, and VEGF) in plasma of over 200 patients with all stages of melanoma (stage I thought IV), melanoma in situ, atypical nevi (possible pre-malignant lesions) as well as normal controls (patients with benign nevi). Due to the volume of data (27 assays for over 200 subjects), principal component analysis (PCA) was used to identify patterns (groupings) of cytokine data based on clinical subject classifications. The results suggested that the most significant differences in plasma cytokine levels among the different patient diagnostic categories was detected in the category of patients with stage IV melanoma/metastatic melanoma (circled brown spheres in
FIG. 3 ). Closer analysis of the data indicated that the most significant difference among the cytokines was noted for IL-4 (FIG. 4 , left), the key regulatory cytokine of the Th2 immune response. Statistical comparisons (Student's t-test) of the mean cytokine concentrations between patients with metastatic melanoma (stage IV) versus all others depicted a pattern of predominance of other Th2 cytokines in addition to IL-4 (IL-10, IL-13, IL-5, eotaxin, IL-9) in patients with stage IV melanoma (FIG. 4 , right). For all of these cytokines, the pattern of plasma concentrations across stages of melanoma was similar to that of IL-4 (no significant changes among any of the patient cohorts except stage IV melanoma). These results are complementary to reports suggesting an increased frequency of Th2 cells in the blood of patients with advanced cancer (as well as chronic infections) relative to normal controls (Inagaki et al., Int. J. Cancer, 118(12):3054-61 (2006); Matsuda et al., Dis. Colon Rectum, 49(4):507-16 (2006); Agarwal et al., Cancer Immunol. Immunother., 55(6):734-43 (2006); and Kumar et al., Oncol. Rep., 15(6):1513-6 (2006)). - Preliminary analysis of three random samples from patients with stage IV melanoma also demonstrated an increased frequency of Th2 cells. These data support the hypothesis of the presence of a state of Th2 mediated chronic inflammation in patients with metastatic melanoma and offer an explanation to the observed state of systemic immune dysfunction (e.g., inability to generate effective immunity following vaccination with cancer vaccines) emulating other clinical conditions characterized by Th2 driven systemic chronic inflammation.
- PBMC isolated from patients with benign nevi, atypical (including dysplastic) nevi, as well as patients with in situ, stage I, II, III or IV melanoma were analyzed by flow cytometry to determine the frequencies of T, NK, and dendritic (DC) cell subsets. There were not significant differences in frequencies of T-cell among stages of melanoma as determined by numbers of CD3, CD4 or CD8 positive T-cells (Table 2). Similarly, no significant differences were found in activated T-cells (CD3/CD69), total NK cells (CD16/56+, CD3−), or most DC subset parameters. As patients with stage IV melanoma appeared to differ significantly from all others with regard to plasma cytokine profiles, the cell subset analysis of patients with stage IV melanoma were compared relative to all others. The analysis revealed no significant differences among most parameters (Table 3) with the following exceptions: (a) the frequency of naïve T-cells (CD3/CD62L+) as well as activated DC (CD11c/CD83+) were significantly less in patients with stage IV melanoma; and (b) the frequency of tetramer positive CTL for gp100 and tyrosinase (but not MART-1 or CMV and EBV) were increased in patients with stage IV melanoma. Due to lack of available biospecimens, Th1 and Th2 enumeration could not be performed. These data suggested that there appeared to be some level of “immune activation” in patients with metastatic melanoma that was different from all other cohorts, and this was consistent with a state of Th2 mediated “chronic inflammation.”
-
TABLE 2 Square root averages of cell subsets with the 95% confidence interval (parenthesis). The p-value represents the comparison across all stages of disease. Benign Atypical In-Situ Stage I Stage II Stage III Stage IV Variable (N = 22) (N = 21) (N = 27) (N = 27) (N = 12) (N = 13) (N = 32) p-value Sqrt CD3 4.78 5.46 5.74 5.29 5.50 5.63 4.94 0.20 (4.20, 5.37) (4.88, 6.04) (5.23, 6.25) (4.78, 5.80) (4.79, 6.20) (4.94, 6.32) (3.92, 5.96) Sqrt CD3/4 3.97 4.54 4.57 4.40 4.51 4.64 4.00 0.59 (3.41, 4.53) (3.98, 5.09) (4.08, 5.06) (3.91, 4.89) (3.84, 5.19) (3.98, 5.31) (3.02, 4.97) Sqrt CD3/8 2.43 2.82 3.17 2.64 2.60 2.93 2.56 0.20 (1.96, 2.89) (2.36, 3.28) (2.77, 3.57) (2.23, 3.04) (2.04, 3.16) (2.39, 3.48) (1.76, 3.37) Sqrt 2.92 2.77 2.91 2.28 2.67 3.15 1.13 0.17 CD3/62L (2.23, 3.60) (2.09, 3.45) (2.31, 3.50) (1.69, 2.88) (1.84, 3.49) (2.35, 3.96) (0.00, 2.32) Sqrt CD3/69 0.40 0.54 0.56 0.50 0.50 0.48 0.49 0.95 (0.18, 0.61) (0.32, 0.75) (0.37, 0.75) (0.31, 0.69) (0.24, 0.76) (0.23, 0.74) (0.12, 0.87) Sqrt 3.03 3.54 3.44 3.30 3.18 3.43 3.48 0.55 CD3/16 + 56 (2.62, 3.44) (3.13, 3.95) (3.08, 3.80) (2.94, 3.66) (2.68, 3.68) (2.95, 3.92) (2.77, 4.20) Sqrt 11c+/14− 1.88 2.15 1.89 1.89 2.00 1.98 1.64 0.66 (1.59, 2.17) (1.86, 2.44) (1.64, 2.14) (1.64, 2.15) (1.65, 2.35) (1.64, 2.32) (1.13, 2.14) Sqrt 11c+/14+ 2.74 3.07 2.60 3.11 2.86 3.50 2.80 0.33 (2.18, 3.3) (2.52, 3.63) (2.11, 3.09) (2.62, 3.60) (2.18, 3.54) (2.84, 4.16) (1.82, 3.77) Sqrt 11c+/DR 0.80 0.91 0.74 0.87 0.74 0.86 0.78 0.48 (0.65, 0.95) (0.77, 1.06) (0.61, 0.87) (0.74, 1.00) (0.56, 0.92) (0.69, 1.03) (0.53, 1.03) Sqrt 11c+/DR+ 1.06 1.21 1.06 1.05 1.11 1.01 1.33 0.45 (0.88, 1.23) (1.04, 1.39) (0.90, 1.21) (0.90, 1.20) (0.90, 1.32) (0.81, 1.22) (1.03, 1.64) Sqrt 11c/80 0.22 0.26 0.24 0.24 0.21 0.25 0.18 0.46 (0.17, 0.26) (0.21, 0.30) (0.20, 0.28) (0.20, 0.28) (0.15, 0.26) (0.20, 0.31) (0.10, 0.25) Sqrt 11c/83 0.21 0.27 0.24 0.24 0.18 0.19 0.1 0.18 (0.15, 0.28) (0.21, 0.33) (0.19, 0.30) (0.18, 0.29) (0.11, 0.26) (0.12, 0.26) (0.00, 0.21) Sqrt 11c/86 0.70 0.82 0.71 0.76 0.70 0.75 0.74 0.84 (0.56, 0.83) (0.68, 0.95) (0.59, 0.82) (0.64, 0.87) (0.53, 0.86) (0.59, 0.91) (0.50, 0.97) Sqrt DR/40 0.54 0.74 0.55 0.62 0.60 0.61 0.82 0.36 (0.37, 0.70) (0.58, 0.91) (0.40, 0.69) (0.47, 0.76) (0.40, 0.80) (0.42, 0.80) (0.54, 1.11) -
TABLE 3 p-value Cytokine (stage IV vs all other) IL-4 1.73 × 10−12 RANTES (CCL5) 6.17 × 10−06 IL-10 5.29 × 10−05 Eotaxin (CCL11) 8.31 × 10−05 IP-10 (CXCL10) 0.0007 IL-13 0.002 IL-12p70 0.005 IL-7, IL-9 0.009 VEGF, MIB-1b 0.02 (CCL4) GM-CSF 0.03 IL-5 0.05 IL-15, TNFa, MIP-1a, >0.05 FGF, IL-2, G-CSF, IL- 8, IL-6, IFN-g, MCP-1, IL-17, PDGF, IL-1ra, IL-1b - The emerging data seemed to suggest that patients with stage IV melanoma, unlike all other patients with earlier stages of melanoma (or healthy controls), existed in a state of systemic Th2 dominance with some evidence of cellular immune activation in peripheral blood (increased frequencies of tumor specific CTL and decreased frequencies of naïve T cells). This immune homeostasis profile resembled a state of Th2 dominant “chronic inflammation,” similar to chronic viral infection (Sester et al., Am. J. Transplant, 5(6):1483-89 (2005)). A reflection of the chronic inflammatory state of chronic viral infection as well as metastatic melanoma is an increase in peripheral blood PD-1+ (exhausted) T-cells (Wong et al., Int. Immunol., 19:1223-34 (2007)). The same was found to be true in the patient cohort of stage IV melanoma patients compared to healthy controls (
FIG. 5 a). This was further supported by functional assessment of antigen specific CTL, revealing a significant reduction in the frequency of functional recall antigen (CMV495-503) specific CTL in patients with stage IV melanoma versus healthy volunteers (FIG. 5 b). Less than 5% of tumor antigen specific, PBMC derived, tetramer positive CTL (MART-1) were capable of intracellular IFN-γ synthesis suggesting immune tolerance. - The plasma cytokine profiling data comparing patients across all stages of melanoma suggested that the greatest differences in the measured parameters occurred in the setting of metastatic melanoma (
stage 4 disease). Therefore, it was hypothesized, that the presence of visible metastatic disease was in some way responsible for the detected Th2 cytokine dominance in these patients and was likely the result of molecules produced by the tumor and/or its interaction with surrounding immune cells. To that end, the mRNA expression profile of 24 biopsy specimens of human metastatic melanoma was analyzed looking for up-regulation of expression of known regulatory molecules of immunity (cytokines and chemokines). The mRNA was extracted from frozen sections in areas that by H&E staining appeared to contain pure tumor tissue (devoid of necrotic tissue, stroma or lymphocytic infiltrates). The RNA was analyzed using an Affymetrix U133 plus 2.0 array. In this experiment, concurrent blood samples were not available from the patients for whom tumor tissues existed. The expression of 23 cytokines (45 probes): IL1a and b, IL-1ra, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12, IL-13, IL-15, IL-17, IFN-γ, CCL5, CCL11, CSF-2, MCP-1, TNF-α, VEGF was analyzed. The objective of the experiment was to determine whether or not the malignancy itself was the source of Th2 cytokines that were detected in plasma. The data revealed that many of the probed cytokines, chemokines, and growth factors are up-regulated in tumor tissue (FIG. 6 a). However, there were no differences in expression of Th1 vs. Th2 cytokines (IFN-γ vs. IL-4, IL-5, IL-10, and IL-13,FIG. 6 b) in tumor tissues suggesting that the observed Th2 cytokine predominance in plasma was not derived from the tumor. However, of the tested cytokines, the most highly/frequently up-regulated transcript in the tumor samples was VEGF (FIG. 6 b). Plasma VEGF levels were significantly higher in metastatic melanoma patients relative to healthy donors, (FIG. 6 c), consistent with published reports (Tas et al., Melanoma Res., 16:405-11 (2006)). Considering the described immune modulatory (down-regulatory) properties of VEGF (Gabrilovich et al., Nat. Med., 2:1096-103 (1996)), it was postulated that tumor derived VEGF could be responsible for the Th2 polarization in patients with stage IV melanoma (away from the normal state of Th1 dominance). - VEGF has been associated with DC polarization towards DC2 leading to Th2 immune responses (e.g., asthma). Likewise, Th2 cytokines (e.g., IL-4, IL-5, and IL-13) have been associated with increased production of VEGF by a range of different cell types including smooth muscle cells. Preliminary data demonstrated that the addition of recombinant human VEGFA to a 2-day culture of normal blood-donor derived PBMC appeared to shift Th polarity away from Th1 and towards Th2 in a dose dependent fashion (
FIG. 7 ). The addition of VEGF to the cell culture favored a reduction of Th cells capable of IFNγ synthesis and TIM-3 expression (Th1) and increased the frequency of Th cells capable of IL-4 synthesis and CD294 surface expression (Th2). The effect on intracellular IL13 production was less pronounced. Thus, it appeared that VEGFA had a direct impact on human PBMC Th1/Th2 polarization in vitro. - In addition, healthy donor PBMC were stimulated with CD3+/CD28+ expander micro-beads for 3 days with increasing concentrations (1 pg/mL-16 pg/mL) of recombinant VEGF and assessed intracellular cytokine production of IL-4 (Th2 cytokine) and IFN-γ (Th1 cytokine) in CD4+ T-cells at the end of in vitro culture (
FIG. 8 a). The data demonstrated that increasing concentrations of VEGF resulted in a dose-dependent reversal of the relative ratio of Th1 to Th2 cells in favor of Th2. Increased concentrations of VEGF were associated with a decrease in the number of Th1 cells (CD4+/IFNγ+) with an associated reciprocal increase in Th2 cells (CD4+/IL-4+). The polarizing effects of VEGF were lost if the assay was performed with purified CD4 cells only (FIG. 8 b) suggesting that the observed Th polarization effect of VEGF is indirect, likely mediated by other PBMC. The addition of 10 μg/mL of IL-12 to the culture containing 16 pg/mL of VEGF prevented the shift in T-helper polarity from Th1 to Th2; addition of anti-human IL-12 antibody to the stimulated PBMC mimicked the effect of VEGF (FIG. 8 a). These data suggest a possible role for monocyte/macrophages in the PBMC preparation as the mediators of the VEGF induced Th polarization. - To gain further insight into the potential role of tumor produced VEGF on systemic immune homeostasis in vivo in humans with metastatic melanoma, frozen peripheral blood specimens were randomly selected from a recently completed clinical trial (N047a) where patients with metastatic melanoma were treated with chemotherapy (paclitaxel+carboplatin) and a specific anti-VEGFA antibody (bevacizumab). They were analyzed for changes in plasma VEGFA levels and Th1/Th2 polarity as well as frequency of tumor specific CTL (tetramer assay). If VEGF was responsible for Th2 polarization, its suppression using chemotherapy/anti-VEGF therapy would revert the Th1/Th2 balance back to normal (normal ratio is 1:1) and perhaps result in emergence of naturally processed anti-tumor specific CTL. Also, this particular clinical trial was chosen because of: (1) available, appropriately stored biospecimens; and (2) this single arm phase II clinical trial yielded favorable clinical outcomes for patients with metastatic melanoma, suggesting possible clinical relevance of this therapeutic strategy. As illustrated in this example from a single patient (
FIG. 9 ), coincident with the decrease of plasma VEGFA concentrations (as a result of therapy,FIG. 9 , top left), there was an increase in Th1:Th2 ratio away from Th2 and towards Th1 (FIG. 9 , top right) at the same time as the increase in frequency of tumor specific CTL in peripheral blood (reactive against melanoma differentiation antigens: MART-1, gp100 or tyrosinase,FIG. 9 , bottom right). Similar observations were made in two other patient samples. In all three cases, the increases in tumor specific CTL tetramer frequency coincided with a reduction in plasma VEGFA levels to normal levels and a shift in Th polarity away from Th2 and towards Th1. Of note, such an increase in CTL frequency was not observed when analyzing the same immunological parameters from patients with metastatic melanoma treated with chemotherapy alone (nab-paclitaxel+carboplatin) without the anti-VEGF antibody (three patients analyzed from protocol N057e). Of note, the presented data were consistent with a published anecdotal observation from a prior clinical trials demonstrating increase in tumor specific CTL tetramer frequency coincident with reduction of plasma VEGFC levels in patients with metastatic melanoma treated with a thrombospondin-1 analog, ABT-510 (Markovic et al., Am. J. Clin. Oncol., 30(3):303-9 (2007)). Therefore, viewed in the context of the current hypothesis, in addition to the originally postulated anti-tumor and anti-angiogenic goals of paclitaxel/carboplatin/bevacizumab therapy, the effect of chemotherapy (paclitaxel and carboplatin) may also have depleted (lymphodepleted) the pre-existing state of “chronic inflammation”; and the VEGF inhibitor (bevacizumab) may have allowed reconstitution of tumor-specific immunity in a Th1 (not Th2) dominant systemic environment. Thus, it is possible that the additional, unanticipated, immunomodulatory effect of this therapy may have added to the observed therapeutic clinical result. Repeated treatments with lymphodepleting chemotherapy may have also inadvertently lead to ultimate depletion of the beneficial anti-tumor immune response as well, allowing tumor progression. Perhaps, this explains in part the clinical outcomes of protocol N047a demonstrating a dramatic improvement in median progression free survival (from 6 weeks to 6 months) with a not nearly as significant an improvement in overall survival (from 8 months to 12 months). - In summary, preliminary data suggests that patients with advanced (metastatic) melanoma exhibit systemic features of Th2-mediated chronic inflammation that appears at least in part mediated by tumor-secreted VEGF (
FIG. 10 ). This state of chronic inflammation effectively dampens spontaneously developed anti-tumor CTL immune responses and significantly reduces the efficacy of de novo immunization efforts with cancer vaccines/immune modulation in this patient population. Evidence exists that the observed Th2/VEGF pathway could be self sustaining and exists in both physiologic (pregnancy) as well as other pathologic states (e.g., asthma) in humans. Disruption of the Th2 driven systemic chronic inflammation in patients with advanced melanoma (and possibly other malignancies) and reconstitution of effective immunity (Th1 dominance) could potentially translate into effective therapy with clinically meaningful results. Therefore, an improved understanding of this mechanism of tumor mediated immune dysfunction/tumor progression as a function of Th2-mediated chronic inflammation could yield therapeutic targets for cancer therapy with agents already in clinical development for Th2 mediated disorders (e.g., anti-IL-4 antibody). - The identified cytokine profiles of plasma suggests the existence of a Th2 dominant systemic immune environment in the blood of patients with metastatic melanoma. The analysis of the cellular/functional counterpart of the immune response in these patients across all stages of melanoma remains unknown. To confirm the hypothesis of Th2 dominant systemic immunity, the existence of reciprocal, Th2 polarized, changes in the cellular immune response in these patients across stages of melanoma that will correlate with the described changes in plasma cytokine and VEGF concentrations is determined To address this, one can (a) enumerate Th1, Th2 and Treg cells across stages of melanoma; (b); analyze the numbers and functional/differentiation state of circulating DC (DC1/DC2) across stages of melanoma (DC2 driven Th2 polarization) and (c) analyze the functional status (active vs. tolerant) of both tumor-specific (e.g. MART-1, gp100, tyrosinase) and recall antigen specific (EBV, CMV) CTL in the HLA-A2+ subset of patients, across stages of melanoma. These data can be combined with the existing data on plasma cytokine levels and correlated looking for patterns of Th1 vs. Th2 cytokine/cellular profiles across patients and in relation to plasma VEGF levels.
- Enumeration of Th1, Th2 and Treg cells across stages of melanoma. In order to assess whether the Th2 cytokine predominance in plasma of patients with metastatic melanoma is truly a reflection of a systemic immune polarization towards Th2 driven chronic inflammation, one can ascertain the expected corresponding changes in the frequencies of circulating Th cell subsets across disease stage using frozen PMBC samples corresponding to plasma cytokine samples described above. The available frozen PBMC can be thawed and analyzed for the relative numbers of Th1, Th2 and Treg. CD4 cells can be isolated from thawed PBMC specimens using paramagnetic beads coated with anti-CD4 (Dynal, Oslo, Norway), and they can be incubated with mouse-anti-human CD3/CD28 coated “stimulator” micro-beads (R and D Systems Minneapolis, Minn.) for 6 hours in the presence of 1 ug/ml brefeldin A, (Sigma Aldrich, St Louis, Mo.). After stimulation, the cells can be fixed, permeablized, and stained with APC conjugated mouse anti-human CD4 (Becton Dickinson, San Jose, Calif.) and FITC conjugated mouse anti-human IFNγ and anti-IL-13 (R and D Systems Minneapolis, Minn.). The stained samples can be analyzed by flow cytometry (FACScan and Cellquest software (Becton-Dickinson, San Jose, Calif.). The results can indicate the percentage of IFNγ positive/IL-13 (or IL-4) negative (Th1) and IFNγ negative/IL-13 (or IL-4) positive (Th2) helper T cells.
- GLP validation of anti-CD294 and anti-TIM-3 cell-surface immunostaining can be completed for the distinction of Th2 vs. Th1 cells in ex vivo (unstimulated) frozen PBMC, respectively. Preliminary data suggested that CD4/CD294 positive Th2 cells exclusively produce IL-4 and IL-13 and not IFNγ upon CD3/CD28 stimulation. Conversely, CD4/TIM-3 positive Th1 cells exclusively produced IFNγ and not IL-4 and IL-13 following the same in vitro stimulation (
FIG. 11 ). Once validated, the assay can be standardized and applied to the battery of tests described herein. - Enumeration of Treg can be performed using intracellular staining for FoxP3 of CD4/25 positive lymphocytes Immunophenotyping can be conducted using commercially available monoclonal antibodies (Biolegend; San Diego, Calif.). Samples can be analyzed by flow-cytometry by FACScan® and data processed using Cellquest® software (Becton-Dickinson, Franklin Lakes, N.J.).
- Assessment of peripheral blood DC subset (DC1/DC2) and activation/differentiation status. In order to ascertain whether or not the state of Th2 driven systemic chronic inflammation is primarily a function of systemic DC polarization towards DC2 (from DC1) leading to HTL polarization from Th1 to Th2 in patients with advanced melanoma, one can quantify the relative numbers and functional states of DC1 and DC2 in patients with melanoma across stages of disease. These data can be analyzed in conjunction with corresponding plasma cytokine/VEGF and Th subset data (above). Therefore, the available, frozen PBMC corresponding to the plasma cytokine samples described above can be thawed and analyzed for the relative numbers of DC subsets defined by expression of CD11c+/CD123-(DC1), and CD11c-/CD123+ (DC2). Each subset can be analyzed for surface expression of co-stimulatory molecules (CD80, 83, 86) Immunophenotyping can be conducted using commercially available monoclonal antibodies (BD Pharmingen; San Jose, Calif.). Samples can be analyzed by flow-cytometry by FACScan® and data processed using Cellquest® software (Becton-Dickinson, Franklin Lakes, N.J.).
- Analysis of the functional status (active/tolerant) of tumor specific (MART-1, gp100, tyrosinase) and recall antigen specific (EBV, CMV) CTL in the HLA-A2+ subset of patients across stages of melanoma. Available frozen PBMC for the same cell repository as described herein can be analyzed for the frequency and functional capacity (exhausted, tolerant vs. non-tolerant subsets) of tumor specific and recall antigen specific CTL. If the hypothesis of tumor mediated, Th2-driven chronic inflammation is correct, the predominant phenotype of the tumor (and recall) antigen specific CTL can be one of tolerance (inability to synthesize intracellular IFNγ upon congnant stimulation with tumor-specific peptides) and exhaustion (expression of PD-1). The latter has already been suggested to be true (Rosenberg et al., J. Immunol., 175(9):6169-76 (2005)) Immunophenotyping of PBMC can be performed using tetramers for melanoma differentiation antigen specific, HLA-A2 congnant peptides (MART-127-35, gp100209-217 and tyrosinase368-376) as well as A2 cognant peptides of EBV and CMV (Beckman Coulter, San Diego, Calif.). For tetramer analysis, thawed PBMC can be stained with FITC conjugated anti-CD8, PC5 conjugated anti-human CD4, CD14 and CD19, and conjugated HLA-A2 tetramers containing peptides from CMV, EBV (controls), MART-127-35, gp100209-217 and tyrosinase368-376. Samples can be analyzed by flow-cytometry and data processed using Cellquest® software (Becton-Dickinson, Franklin Lakes, N.J.). Gates can be set on lymphocytes that were CD4, CD14, and CD19 (PC5) negative and CD8 (APC) positive. Ongoing quality assurance (QA) data suggests an inter-assay variability with a coefficient of variation (CV) below 5%. Standard control samples can be run alongside all experiments. If the standard control samples generate results beyond ±2SD of the mean, all assay results can be rejected and the experiment repeated.
- Functional analysis of tetramer positive CTL can be performed in patient samples demonstrating tetramer frequencies of at least 0.1% to melanoma differentiation or recall antigens. One can proceed to ascertain the ability of tetramer positive CTL to synthesize interferon-y (IFNγ) upon stimulation with cognate peptide presented in the context of HLA-A2 and anti-CD28 co-stimulation using artificial antigen presenting cell (aAPC) stimulation. The details of this method are described elsewhere (Markovic et al., Clin. Exp. Immunol., 145(3):438-47 (2006)). In brief, previously frozen patient PBMC can be thawed in batches, labeled with PE conjugated tetramers (Beckman Coulter, San Diego, Calif.), and stimulated for 6 hours, in the presence of 1 mg/mL brefeldin A, (Sigma Aldrich, St Louis, Mo.) with pararamagnetic beads (Dynal, Oslo, Norway) coated with peptide loaded HLA-A2 (Beckman Coulter San Diego, Calif.) and mouse anti-human CD28 (R and D Systems Minneapolis, Minn.). After stimulation, the cells can be fixed, permeablized, and stained with APC conjugated mouse anti-human CD8 (Becton Dickinson, San Jose, Calif.) and FITC conjugated mouse anti-human IFN-gamma (R and D Systems Minneapolis, Minn.). The stained samples can be analyzed by flow cytometry (FACScan and Cellquest software (Becton-Dickinson, San Jose, Calif.). The results can indicate the percentage of tetramer positive CTL able vs. unable to synthesize intracellular IFNγ. Ongoing QA data suggests an inter-assay variability with a CV of below 9%. Standard control samples can be run alongside all experiments. If the standard control samples generate results beyond ±2SD of the mean, all assay results can be rejected, and the experiment repeated.
- Laboratory data summary and statistical analysis. All blood samples were registered, collected, processed and annotated. Sample break down per diagnostic category is described in Table 4.
-
TABLE 4 Summary of available biospecimens (frozen PBMC and plasma) Clinical diagnostic category Benign Atypical In Situ Stage I Stage II Stage III Stage IV nevi nevi melanoma melanoma melanoma melanoma melanoma Available frozen 26 16 35 36 12 16 30 PBMC with already available corresponding plasma cytokine data Additional available frozen PBMC and frozen plasma as of May 1, 2008 Therapeutic 0 0 0 0 0 44 198 clinical trials Melanoma blood 10 0 22 43 38 209 421 and tissue bank (MC997 g) - The effects of melanoma-specific therapeutic interventions on the VEGF/Th2 state of tumor-induced chronic inflammation can be examined using peripheral blood biospecimens from patients with stage IV malignant melanoma enrolled on ongoing or completed clinical trials for changes in a range of immune parameters with a primary focus on Th1/Th2 balance and functional tumor specific CTL immunity. The trials are listed in the Table 5. The patients enrolled into these trials had a blood specimen collected before initiation of therapy and after one cycle of treatment. This can provide data on the immediate impact of therapy on our VEGF/Th2/immune parameters of interest.
-
TABLE 5 Summary of available biospecimens from therapeutic clinical trials for stage IV melanoma Study Number of patients enrolled number Treatment regimen (accrual as of May 1, 2008) N0377 RAD0012 53 (completed) N047a Paclitaxel + Carboplatin + 53 (completed) Bevacizumab1 N057e Abraxane + Carboplatin 74 (completed) MC057f Temozolomide 86 (12) Paclitaxel + Carboplatin 86 (0) N0675 RAD0012 + Temozolomide 43 (6) N0775 Abraxane + Carboplatin + 43 (0) Bevacizumab1 Temozolomide + Bevacizumab 43 (0) 1Humanized anti-VEGFA antibody; 2rapamycin analog, inhibitor of mTOR (down-regulation of VEGF synthesis) - For each individual patient, the biospecimens collected before initiation of therapy and after one cycle of treatment can be used. This can provide data on the immediate impact of therapy on VEGF/Th2/immune parameters of interest. Additional testing for later time-points can be pursued only if justified by the initial analysis suggesting beneficial changes in the studied immune parameters. This can allow one to gain insight into the effects of a broad range of clinical interventions on immune homeostasis using an available (but limited) resource of biospecimens and only pursue further analysis if justified by the generated data.
- Laboratory analyses of the stored PBMC can include the same assays described above and can also include: (a) PBMC immunophenotyping for immune cell subset analysis; and (b) plasma cytokine profiling.
- PBMC immunophenotyping for immune cell subset analysis. Pre and post-treatment frozen PBMC biospecimens can be analyzed for the “global” impact of therapy on immune cell subsets. One can analyze the relative numbers of T, B, NK cells, monocytes and DC, and their activation status using commercially available monoclonal antibodies directed at the following antigens: CD3, CD4, CD8, CD11c, CD14, C16, CD19, CD20, CD25, CD45RA/RO, CD56, CD69, CD63L, CD80, CD83, CD86, CD123, DR, foxP3 (BD Pharmingen; San Jose, Calif.) Immunophenotyping can be performed using manufacturer's instruction in batch samples of the same patients analyzed on the same day. The stained samples can be analyzed by flow cytometry (FACScan and Cellquest software (Becton-Dickinson, San Jose, Calif.). PBMC isolated from patients prior to B7-DC XAb and 15 days after antibody treatment can be assayed. Changes in the numbers of cells bearing the lymphocyte markers in pared comparisons for patients prior to B7-DC XAb treatment can be used to ascertain antibody treatment effects.
- Plasma cytokine profiling. In order to complement the PBMC derived cellular immunity analyses, one can add plasma cytokine measurements in the same samples (paired PBMC and plasma testing). To that end, one can profile the serum cytokine changes as a result of specific therapy for all available specimens before and after treatment. The BioRad human 27-plex cytokine panel can be used (Cat # 171-A11127, Bio-Rad, San Diego Calif.) for the measurements of plasma concentrations of IL-1β, IL-1rα, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12 (p70), IL-13, IL-15, IL-17, basic FGF, Eotaxin, G-CSF, GM-CSF, IFN-γ, IP-10, MCP-1, MIP-1α, MIP-1β, PDGF, RANTES, TNF-α, and VEGF. The assay can be performed as per the manufacturer's directions. Briefly, 100 μL of Bio-Plex assay buffer can be added to each well of a MultiScreen MABVN 1.2 μm microfiltration plate followed by the addition of 50 μL of the multiplex bead preparation. Following washing of the beads with the addition of 100 μL of wash buffer, 50 μL the samples or the standards can be added to each well and incubated with shaking for 30 minutes at room temperature. The plasma (1:3 dilution) and standards can be diluted using the Bio-Plex human serum diluent kit and plated in duplicate. Standard curves can be generated with a mixture of 27 cytokine standards and eight serial dilutions ranging from 0-32,000 pg/mL. The plate can then be washed 3 times followed by incubation of each well in 25 μL of pre-mixed detection antibodies for 30 minutes with shaking. The plate can further be washed and 50 μL of streptavidin solution were added to each well and incubated for 10 minutes at room temperature with shaking. The beads can be given a final washing and resuspension in 125 μL of Bio-Plex assay buffer. Cytokine levels in the sera can be quantified by analyzing 100 μL of each well on a Bio-Plex using Bio-Plex Manager software version 4.0. Normal values for plasma cytokine concentrations were generated by analyzing 30 plasma samples from healthy donors (blood donors at the Mayo Clinic Dept. of Transfusion Medicine). A set of five normal plasma samples (standards) can be run along side all batches of plasma analysis. If the cytokine concentrations of the “standard” samples differ by more than 20%, results can be rejected, and the plasma samples re-analyzed.
- To better understand the clinical relevance of these differences in immune homeostasis and study the kinetics of their evolution in humans as they develop clinically detectable metastatic cancer (melanoma), one can perform a prospective clinical trial in which patients with surgically resected metastatic melanoma (and in a state of chronic inflammation) undergo complete resection of their tumors as part of their clinical care and are subsequently followed at regular time-intervals until tumor relapse. It is hypothesize that following surgical resection, the state of systemic “chronic inflammation” will resolve. These patients can then be followed at regular intervals (every 2 months), and their blood analyzed for emergence of Th2-mediated chronic inflammation, until clinical tumor relapse/recurrence of metastatic melanoma (approximately 50% of patients will relapse within 18 months of surgery). This study will depict the time-sequence and thresholds of systemic changes in immune homeostasis (chronic inflammation) as they evolve towards the development of relapsed metastatic melanoma. Sufficient blood specimens (100 mL every 2 months) can be collected to allow complete analysis as well as provide some additional material for further testing (if necessary). The clinical trial can be powered based on the inter-patient variability of the most prominent immune abnormality in patients with metastatic melanoma (e.g. variability of plasma IL-4 concentrations) determined herein. If successful, these data can clinically validate the changes in immune homeostasis as they impact the natural history of metastatic melanoma and describe potential targets for future therapy. Additionally, as all patients on this study will undergo surgical resection of metastatic melanoma as well as undergo concurrent comprehensive immunological testing, their surgical tissues can be processed and preserved for analysis addressing the influence of the tumor on the observed immunological profile (multiple frozen blocks for future immunohistochemical study and mRNA extraction). All tumor tissue can undergo genomic mRNA expression profiling as well as IHC analysis for infiltrating immune cell subsets (funded under separate, existing instruments). These data can correlate the relationship of immunity in the tumor microenvironment with that of systemic immunity in metastatic cancer.
- Study design. The clinical trail can be conducted in the context of the clinical trials program of the Melanoma Study Group of the Mayo Clinic Cancer Center. All patients with the diagnosis of metastatic melanoma that are planned to undergo complete surgical resection of their malignancy can be offered participation in this study. The objective of the study can be to profile the changes in immune homeostasis from pre-surgery, post-surgery and all through the time of clinically detectable tumor relapse. It is hypothesized that the state of VEGF/Th2 driven chronic inflammation can be established pre-surgery, resolved soon after surgery and slowly re-develop in the months prior to clinical tumor relapse.
- For the purposes of this study, patients can be clinically followed in accordance to clinical practice (every 2 months). Patients can be asked to donate 100 mL of blood at each follow-up time point. The blood can be collected, processed, and stored in accordance to existing procedures for immunological testing Immune homeostasis analysis can be conducted in batches to limit inter-assay variability. Specific focus/priority can be given to parameters reflecting Th1/Th2 balance, frequency of functional/tolerant tumor (or recall) antigen specific CTL as well as plasma cytokine and VEGF levels. At the time of clinical relapse, patients can be re-tested, and the tumor biopsied for histologic confirmation. Available tumor tissues can be analyzed for expression of tumor associated antigens (immunohistochemistry for MART-1, gp100 and tyrosinase) as well as tumor infiltrating lymphocytes and compared to the original surgical specimen for each individual patient. In the rare events where patients can undergo another curative surgical resection at the time of relapse, they may continue on study following the outlined follow-up/testing schedule until such a time when a tumor relapse is no longer surgically resectable. See, e.g., Table 6.
- Eligibility Criteria.
- Required Characteristics/Inclusion Criteria:
- 1. HLA-A2+ adult patients (age 18 years) with metastatic malignant melanoma who are planned to undergo complete resection for metastatic disease as part of their regular medical care.
2. The following laboratory values obtained 14 days prior to registration: hemoglobin ≧9.0 g/dL; platelet count ≧75,000/μL; and AST≦3×ULN.
3. Ability to provide informed consent.
4. Willingness to return to clinic for follow-up.
5.ECOG performance status
6. Willingness to participate in the mandatory translational research component of the study. - Contraindications/Exclusion Criteria:
- 1. Uncontrolled or current infection.
2. Known standard therapy for the patient's disease that is potentially curative or proven capable of extending life expectancy.
3. Any of the following prior therapies with interval since most recent treatment: (a) chemotherapy ≦4 weeks prior to registration; or (b) biologic therapy weeks prior to registration.
4. Any of the following as this regimen may be harmful to a developing fetus or nursing child: (a) pregnant women; (b) nursing women; or (c) women of childbearing potential or their sexual partners who are unwilling to employ adequate contraception (condoms, diaphragm, birth control pills, injections, intrauterine device (IUD), surgical sterilization, subcutaneous implants, or abstinence, etc).
5. Known immune deficiency or ongoing immunosuppressive therapy. -
TABLE 6 Test schedule Every 2 months after ≦14 days prior <7 days prior to surgery until surgically At time of Tests and procedures to registration scheduled surgery unresectable relapse2 tumor relapse2 History and exam, X X X weight, performance status Vital signs X X Disease evaluation X X X (clinical/imaging) Hematology group X X X WBC, ALC, ANC, Hgb, platelets Chemistry group X X X AST, LDH, Alk Phos, Creat, K, Na, LDH Immunology studies XR XR XR HLA typing XR, Tumor typing for XR XR MART1, gp100 and tyrosinase; profile of infiltrating lymphocytes Serum pregnancy test1 XR 1Only for women of child-bearing age; 2if a patient has a melanoma relapse that is surgically completely resectable, they may continue on study with the same follow-up/testing schedule until relapse is no longer surgically resectable; Rresearch funded - CD4 T cells (Th1 and Th2 CD4 T cells) derived from human PBMC were incubated in vitro with varying concentrations of VEGFA (rhVEGFA; 10 ng/mL, 50 ng/mL, and 200 ng/mL) or TGFβ (rhTGFβ; 10 ng/mL, 50 ng/mL, and 200 ng/mL). After six hours of incubation at 37° C., the ratio of Th1 vs. Th2 (Th1/Th2) was determined Both VEGFA and TGFβ exhibited a similar effect on Th1/Th2 polarity in human PBMC derived CD4 cells (
FIG. 12 ). - CD4 T cells (Th1 and Th2 CD4 T cells) derived from human PBMC were incubated in vitro with VEGFA alone (1 ng/mL, 5 ng/mL, 10 ng/mL, 100 ng/mL, or 1000 ng/mL) or VEGFA (100 ng/mL) plus an anti-TGFβ antibody (1 ng/mL, 10 ng/mL, 100 ng/mL, 1 μg/mL, 5 μg/mL, or 10 μg/mL). The anti-TGFβ antibody was obtained from Genzyme Corp. (Cambridge, Mass.). Untreated cells (media only) as well as cells exposed to Th1 or Th2 favorable in vitro conditions were used as controls. After six hours of incubation at 37° C., the ratio of Th1 vs. Th2 (Th1/Th2) was determined. The presence of anti-TGFβ antibodies reversed the Th1/Th2 modulation of VEGFA in vitro, suggesting that the observed VEGF effect in these cells may be TGFβ mediated (
FIG. 13 ). - It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.
Claims (15)
1. A method for treating a mammal having cancer, said method comprising:
(a) administering to said mammal an anti-chronic inflammation treatment under conditions wherein the level of global chronic inflammation in said mammal is reduced, and
(b) administering to said mammal a cancer treatment agent under conditions wherein the presence of said cancer is reduced.
2. The method of claim 1 , wherein said mammal is a human.
3. The method of claim 1 , wherein said cancer is melanoma.
4. The method of claim 1 , wherein said cancer is stage IV melanoma.
5. The method of claim 1 , wherein said anti-chronic inflammation treatment comprises chemotherapy, radiation, an anti-IL-4 agent, an anti-IL-13 agent, or a steroid treatment.
6. The method of claim 1 , wherein said cancer treatment agent is a cancer vaccine.
7. The method of claim 1 , wherein said cancer vaccine is a MART-1, gp100, or survivin cancer vaccine.
8. The method of claim 1 , wherein the period of time between the last administration of said anti-chronic inflammation treatment and the first administration of said cancer treatment agent is between two weeks and six months.
9. A method for treating a mammal having cancer, said method comprising:
(a) administering to said mammal an anti-TGFβ antibody under conditions wherein the level of global chronic inflammation in said mammal is reduced, and
(b) administering to said mammal a cancer treatment agent under conditions wherein the presence of said cancer is reduced.
10. The method of claim 9 , wherein said mammal is a human.
11. The method of claim 9 , wherein said cancer is melanoma.
12. The method of claim 9 , wherein said cancer is stage IV melanoma.
13. The method of claim 9 , wherein said cancer treatment agent is a cancer vaccine.
14. The method of claim 9 , wherein said cancer vaccine is a MART-1, gp100, or survivin cancer vaccine.
15. The method of claim 9 , wherein the period of time between the last administration of said anti-TGFβ antibody and the first administration of said cancer treatment agent is between two weeks and six months.
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US12/979,105 US20110150902A1 (en) | 2008-07-03 | 2010-12-27 | Treating cancer |
US14/051,849 US9387244B2 (en) | 2008-07-03 | 2013-10-11 | Methods for reducing global chronic inflammation and the presence of melanoma |
US15/179,392 US10287344B2 (en) | 2008-07-03 | 2016-06-10 | Methods for reducing global chronic inflammation and the presence of melanoma |
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US7820308P | 2008-07-03 | 2008-07-03 | |
PCT/US2009/049511 WO2010003057A2 (en) | 2008-07-03 | 2009-07-02 | Treating cancer |
US12/979,105 US20110150902A1 (en) | 2008-07-03 | 2010-12-27 | Treating cancer |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US2009/049511 Continuation-In-Part WO2010003057A2 (en) | 2008-07-03 | 2009-07-02 | Treating cancer |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US14/051,849 Continuation US9387244B2 (en) | 2008-07-03 | 2013-10-11 | Methods for reducing global chronic inflammation and the presence of melanoma |
Publications (1)
Publication Number | Publication Date |
---|---|
US20110150902A1 true US20110150902A1 (en) | 2011-06-23 |
Family
ID=41466606
Family Applications (3)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US12/979,105 Abandoned US20110150902A1 (en) | 2008-07-03 | 2010-12-27 | Treating cancer |
US14/051,849 Active US9387244B2 (en) | 2008-07-03 | 2013-10-11 | Methods for reducing global chronic inflammation and the presence of melanoma |
US15/179,392 Active US10287344B2 (en) | 2008-07-03 | 2016-06-10 | Methods for reducing global chronic inflammation and the presence of melanoma |
Family Applications After (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US14/051,849 Active US9387244B2 (en) | 2008-07-03 | 2013-10-11 | Methods for reducing global chronic inflammation and the presence of melanoma |
US15/179,392 Active US10287344B2 (en) | 2008-07-03 | 2016-06-10 | Methods for reducing global chronic inflammation and the presence of melanoma |
Country Status (3)
Country | Link |
---|---|
US (3) | US20110150902A1 (en) |
EP (2) | EP3266453A1 (en) |
WO (1) | WO2010003057A2 (en) |
Cited By (19)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20120009203A1 (en) * | 2010-06-29 | 2012-01-12 | Anahid Jewett | Depletion of cancer stem cells |
US9387244B2 (en) | 2008-07-03 | 2016-07-12 | Mayo Foundation For Medical Education And Research | Methods for reducing global chronic inflammation and the presence of melanoma |
US9427477B2 (en) | 2011-05-09 | 2016-08-30 | Mayo Foundation For Medical Education And Research | Cancer treatments |
US9757453B2 (en) | 2012-10-01 | 2017-09-12 | Mayo Foundation For Medical Education And Research | Nanoparticle complexes of anti-CD20 antibodies, albumin and paclitaxel |
US9763982B2 (en) | 2010-06-29 | 2017-09-19 | The Regents Of The University Of California | Depletion of cancer stem cells |
US10213513B2 (en) | 2014-06-16 | 2019-02-26 | Mayo Foundation For Medical Education And Research | Treating myelomas |
US10300016B2 (en) | 2014-10-06 | 2019-05-28 | Mayo Foundation For Medical Education And Research | Carrier-antibody compositions and methods of making and using the same |
US10561726B2 (en) | 2015-10-06 | 2020-02-18 | Vavotar Life Sciences LLC | Methods of treating cancer using compositions of antibodies and carrier proteins with antibody pretreatment |
US10618969B2 (en) | 2016-04-06 | 2020-04-14 | Mayo Foundation For Medical Education And Research | Carrier-binding agent compositions and methods of making and using the same |
US11160876B2 (en) | 2016-09-01 | 2021-11-02 | Mayo Foundation For Medical Education And Research | Methods and compositions for targeting t-cell cancers |
US11241387B2 (en) | 2015-08-18 | 2022-02-08 | Mayo Foundation For Medical Education And Research | Carrier-binding agent compositions and methods of making and using the same |
US11305020B2 (en) | 2016-03-21 | 2022-04-19 | Mayo Foundation For Medical Education And Research | Methods for reducing toxicity of a chemotherapeutic drug |
US11311631B2 (en) | 2016-09-06 | 2022-04-26 | Mayo Foundation For Medical Education And Research | Paclitaxel-albumin-binding agent compositions and methods for using and making the same |
US11351254B2 (en) | 2016-02-12 | 2022-06-07 | Mayo Foundation For Medical Education And Research | Hematologic cancer treatments |
US11427637B2 (en) | 2016-09-06 | 2022-08-30 | Mayo Foundation For Medical Education And Research | Methods of treating PD-L1 expressing cancer |
US11548946B2 (en) | 2016-09-01 | 2023-01-10 | Mayo Foundation For Medical Education And Research | Carrier-PD-L1 binding agent compositions for treating cancers |
US11571469B2 (en) | 2016-01-07 | 2023-02-07 | Mayo Foundation For Medical Education And Research | Methods of treating cancer with interferon wherein the cancer cells are HLA negative or have reduced HLA expression |
US11590098B2 (en) | 2016-09-06 | 2023-02-28 | Mayo Foundation For Medical Education And Research | Methods of treating triple-negative breast cancer using compositions of antibodies and carrier proteins |
US11878061B2 (en) | 2016-03-21 | 2024-01-23 | Mayo Foundation For Medical Education And Research | Methods for improving the therapeutic index for a chemotherapeutic drug |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2011153224A2 (en) * | 2010-06-02 | 2011-12-08 | Genentech, Inc. | Diagnostic methods and compositions for treatment of cancer |
Citations (18)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4350687A (en) * | 1980-02-10 | 1982-09-21 | Research Corporation | Platelet derived cell growth factor |
US5728541A (en) * | 1996-07-12 | 1998-03-17 | Precision Therapeutics, Inc. | Method for preparing cell cultures from biologial specimens for chemotherapeutic and other assays |
US6054297A (en) * | 1991-06-14 | 2000-04-25 | Genentech, Inc. | Humanized antibodies and methods for making them |
US6416967B2 (en) * | 1996-07-12 | 2002-07-09 | Precision Therapeutics, Inc. | Method of using multicellular particulates to analyze malignant or hyperproliferative tissue |
US6417060B2 (en) * | 2000-02-25 | 2002-07-09 | Borealis Technical Limited | Method for making a diode device |
US20020111362A1 (en) * | 1999-10-15 | 2002-08-15 | Joseph Rubinfeld | Inhibition of abnormal cell proliferation with camptothecin and combinations including the same |
US6537579B1 (en) * | 1993-02-22 | 2003-03-25 | American Bioscience, Inc. | Compositions and methods for administration of pharmacologically active compounds |
US20040005318A1 (en) * | 2002-04-12 | 2004-01-08 | Medarex, Inc. | Methods of treatment using CTLA-4 antibodies |
US20040007601A1 (en) * | 2002-06-26 | 2004-01-15 | Masatoshi Masuda | Valve mechanism for tube-type fluid container |
US20050032699A1 (en) * | 2003-07-25 | 2005-02-10 | Jocelyn Holash | Composition of a VEGF antagonist and an anti-proliferative agent |
US20060165652A1 (en) * | 2002-09-06 | 2006-07-27 | Government Of The United States Of America, Repre- Sented By The Secretary Dhhs | Immunotherapy with in vitro-selected antigen-specific lymphocytes after non-myeloablative lymphodepleting chemotherapy |
US7112409B2 (en) * | 1999-01-29 | 2006-09-26 | Center For Molecular Medicine And Immunology | Method of determining cytokine dosage for myelosuppressive state |
US20070020232A1 (en) * | 2005-04-26 | 2007-01-25 | Eisai Co., Ltd. | Compositions and methods for cancer immunotherapy |
US20100047234A1 (en) * | 2007-03-14 | 2010-02-25 | Markovic Svetomir N | Treating skin cancer |
US20100112077A1 (en) * | 2006-11-06 | 2010-05-06 | Abraxis Bioscience, Llc | Nanoparticles of paclitaxel and albumin in combination with bevacizumab against cancer |
US20100172835A1 (en) * | 2008-12-23 | 2010-07-08 | Burnham Institute For Medical Research | Methods and compositions for synaphically-targeted treatment for cancer |
US20110097340A1 (en) * | 2007-10-22 | 2011-04-28 | Sumant Ramachandra | Fully human anti-vegf antibodies and methods of using |
US8119129B2 (en) * | 2008-08-01 | 2012-02-21 | Bristol-Myers Squibb Company | Combination of anti-CTLA4 antibody with dasatinib for the treatment of proliferative diseases |
Family Cites Families (25)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
SE8801537D0 (en) | 1988-04-26 | 1988-04-26 | Ellco Food Ab | CELL CULTURE MEDIUM AND PROCEDURES FOR ITS PREPARATION |
US5252713A (en) | 1988-09-23 | 1993-10-12 | Neorx Corporation | Polymeric carriers for non-covalent drug conjugation |
US5116944A (en) | 1989-12-29 | 1992-05-26 | Neorx Corporation | Conjugates having improved characteristics for in vivo administration |
US5216130A (en) | 1990-05-17 | 1993-06-01 | Albany Medical College | Complex for in-vivo target localization |
US5260308A (en) | 1991-11-06 | 1993-11-09 | Mayo Foundation For Medical Education And Research | Method to increase permeability of the blood-nerve/brain barriers to proteins |
US5736137A (en) | 1992-11-13 | 1998-04-07 | Idec Pharmaceuticals Corporation | Therapeutic application of chimeric and radiolabeled antibodies to human B lymphocyte restricted differentiation antigen for treatment of B cell lymphoma |
AU735884B2 (en) | 1996-04-26 | 2001-07-19 | Magainin Pharmaceuticals, Inc. | Treatment of carcinomas using squalamine in combination with other anti-cancer agents |
CN100462066C (en) | 1997-06-27 | 2009-02-18 | 美国生物科学有限公司 | Novel formulations of pharmacological agents, method for preparation thereof and method for use thereof |
US6616925B1 (en) * | 1998-04-02 | 2003-09-09 | I.D.M. Immuno-Designed Molecules | Combined preparation for the treatment of neoplasic diseases or of infectious diseases |
US7534866B2 (en) | 2005-10-19 | 2009-05-19 | Ibc Pharmaceuticals, Inc. | Methods and compositions for generating bioactive assemblies of increased complexity and uses |
JP2001072589A (en) | 1999-07-06 | 2001-03-21 | Toagosei Co Ltd | Carcinostatic agent |
JP2006506442A (en) * | 2002-07-09 | 2006-02-23 | ポイント セラピューティクス, インコーポレイテッド | Boroproline compound combination therapy |
US20050043233A1 (en) | 2003-04-29 | 2005-02-24 | Boehringer Ingelheim International Gmbh | Combinations for the treatment of diseases involving cell proliferation, migration or apoptosis of myeloma cells or angiogenesis |
EP1740615B1 (en) * | 2004-03-31 | 2014-11-05 | Genentech, Inc. | Humanized anti-tgf-beta antibodies |
ES2376484T3 (en) * | 2004-04-22 | 2012-03-14 | Eli Lilly And Company | COMBINATION THERAPY FOR THE TREATMENT OF THE C�? NCER. |
AU2005294214A1 (en) | 2004-10-07 | 2006-04-20 | Emory University | Multifunctional nanoparticles conjugates and their use |
US20070166388A1 (en) | 2005-02-18 | 2007-07-19 | Desai Neil P | Combinations and modes of administration of therapeutic agents and combination therapy |
GB0621973D0 (en) | 2006-11-03 | 2006-12-13 | Philogen Spa | Binding molecules and uses thereof |
GB2465940A (en) | 2007-10-01 | 2010-06-09 | Hospital For Sick Children | Neural tumor stem cells and methods of use thereof |
EP3266453A1 (en) | 2008-07-03 | 2018-01-10 | Mayo Foundation for Medical Education and Research | Treating cancer |
EP2321408A4 (en) | 2008-08-04 | 2013-04-17 | Allocure Inc | Mesenchymal stromal cell populations and methods of isolating and using same |
AR078161A1 (en) | 2009-09-11 | 2011-10-19 | Hoffmann La Roche | VERY CONCENTRATED PHARMACEUTICAL FORMULATIONS OF AN ANTIBODY ANTI CD20. USE OF THE FORMULATION. TREATMENT METHOD |
EP2625525A4 (en) | 2010-10-08 | 2014-04-02 | Abraxis Bioscience Llc | Sparc microenvironment signature, plasma sparc, and ldh as prognostic biomarkers in the treatment of cancer |
LT2707030T (en) | 2011-05-09 | 2020-07-10 | Mayo Foundation For Medical Education And Research | Cancer treatments |
EP3967306A1 (en) | 2012-10-01 | 2022-03-16 | Mayo Foundation for Medical Education and Research | Cancer treatments |
-
2009
- 2009-07-02 EP EP17169672.7A patent/EP3266453A1/en not_active Withdrawn
- 2009-07-02 EP EP09774506A patent/EP2310006A4/en not_active Withdrawn
- 2009-07-02 WO PCT/US2009/049511 patent/WO2010003057A2/en active Application Filing
-
2010
- 2010-12-27 US US12/979,105 patent/US20110150902A1/en not_active Abandoned
-
2013
- 2013-10-11 US US14/051,849 patent/US9387244B2/en active Active
-
2016
- 2016-06-10 US US15/179,392 patent/US10287344B2/en active Active
Patent Citations (20)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4350687A (en) * | 1980-02-10 | 1982-09-21 | Research Corporation | Platelet derived cell growth factor |
US6054297A (en) * | 1991-06-14 | 2000-04-25 | Genentech, Inc. | Humanized antibodies and methods for making them |
US6537579B1 (en) * | 1993-02-22 | 2003-03-25 | American Bioscience, Inc. | Compositions and methods for administration of pharmacologically active compounds |
US7678552B2 (en) * | 1996-07-12 | 2010-03-16 | Precision Therapeutics, Inc. | Method for selecting therapeutic agents for cancer treatment |
US6416967B2 (en) * | 1996-07-12 | 2002-07-09 | Precision Therapeutics, Inc. | Method of using multicellular particulates to analyze malignant or hyperproliferative tissue |
US5728541A (en) * | 1996-07-12 | 1998-03-17 | Precision Therapeutics, Inc. | Method for preparing cell cultures from biologial specimens for chemotherapeutic and other assays |
US6933129B1 (en) * | 1996-07-12 | 2005-08-23 | Precision Therapeutics, Inc. | Method for culturing and assaying cells |
US7112409B2 (en) * | 1999-01-29 | 2006-09-26 | Center For Molecular Medicine And Immunology | Method of determining cytokine dosage for myelosuppressive state |
US20020111362A1 (en) * | 1999-10-15 | 2002-08-15 | Joseph Rubinfeld | Inhibition of abnormal cell proliferation with camptothecin and combinations including the same |
US6417060B2 (en) * | 2000-02-25 | 2002-07-09 | Borealis Technical Limited | Method for making a diode device |
US20040005318A1 (en) * | 2002-04-12 | 2004-01-08 | Medarex, Inc. | Methods of treatment using CTLA-4 antibodies |
US20040007601A1 (en) * | 2002-06-26 | 2004-01-15 | Masatoshi Masuda | Valve mechanism for tube-type fluid container |
US20060165652A1 (en) * | 2002-09-06 | 2006-07-27 | Government Of The United States Of America, Repre- Sented By The Secretary Dhhs | Immunotherapy with in vitro-selected antigen-specific lymphocytes after non-myeloablative lymphodepleting chemotherapy |
US20050032699A1 (en) * | 2003-07-25 | 2005-02-10 | Jocelyn Holash | Composition of a VEGF antagonist and an anti-proliferative agent |
US20070020232A1 (en) * | 2005-04-26 | 2007-01-25 | Eisai Co., Ltd. | Compositions and methods for cancer immunotherapy |
US20100112077A1 (en) * | 2006-11-06 | 2010-05-06 | Abraxis Bioscience, Llc | Nanoparticles of paclitaxel and albumin in combination with bevacizumab against cancer |
US20100047234A1 (en) * | 2007-03-14 | 2010-02-25 | Markovic Svetomir N | Treating skin cancer |
US20110097340A1 (en) * | 2007-10-22 | 2011-04-28 | Sumant Ramachandra | Fully human anti-vegf antibodies and methods of using |
US8119129B2 (en) * | 2008-08-01 | 2012-02-21 | Bristol-Myers Squibb Company | Combination of anti-CTLA4 antibody with dasatinib for the treatment of proliferative diseases |
US20100172835A1 (en) * | 2008-12-23 | 2010-07-08 | Burnham Institute For Medical Research | Methods and compositions for synaphically-targeted treatment for cancer |
Non-Patent Citations (2)
Title |
---|
Rao et al. (2006), Cancer, Vol. 106, pp. 375-82. * |
Weber (2007), The Oncologist, Vol. 12, pp.864-872. * |
Cited By (53)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US10287344B2 (en) | 2008-07-03 | 2019-05-14 | Mayo Foundation For Medical Education And Research | Methods for reducing global chronic inflammation and the presence of melanoma |
US9387244B2 (en) | 2008-07-03 | 2016-07-12 | Mayo Foundation For Medical Education And Research | Methods for reducing global chronic inflammation and the presence of melanoma |
US9763982B2 (en) | 2010-06-29 | 2017-09-19 | The Regents Of The University Of California | Depletion of cancer stem cells |
US20120009203A1 (en) * | 2010-06-29 | 2012-01-12 | Anahid Jewett | Depletion of cancer stem cells |
US9427477B2 (en) | 2011-05-09 | 2016-08-30 | Mayo Foundation For Medical Education And Research | Cancer treatments |
US10765741B2 (en) | 2011-05-09 | 2020-09-08 | Mayo Foundation For Medical Education And Research | Methods for treating VEGF-expressing cancer using preformed nanoparticle complexes comprising albumin-bound paclitaxel and bevacizumab |
US10376579B2 (en) | 2012-10-01 | 2019-08-13 | Mayo Foundation For Medical Education And Research | Nanoparticle complexes of albumin, paclitaxel, and anti-VEGF antibody for treatment of cancer |
US10471145B2 (en) | 2012-10-01 | 2019-11-12 | Mayo Foundation For Medical Education And Research | Methods for treating cancer using nanoparticle complexes of paclitaxel, rituximab, and albumin |
US10279035B2 (en) | 2012-10-01 | 2019-05-07 | Mayo Foundation For Medical Education And Research | Nanoparticle complexes of paclitaxel, trastuzumab, and albumin |
US11648311B2 (en) | 2012-10-01 | 2023-05-16 | Mayo Foundation For Medical Education And Research | Cancer treatments |
US10307482B2 (en) | 2012-10-01 | 2019-06-04 | Mayo Foundation For Medical Education And Research | Nanoparticle complexes of paclitaxel, cetuximab, and albumin |
US10668151B2 (en) | 2012-10-01 | 2020-06-02 | Mayo Foundation For Medical Education And Research | Nanoparticle complexes of albumin, paclitaxel, and panitumumab for treatment of cancer |
US10507243B2 (en) | 2012-10-01 | 2019-12-17 | Mayo Foundation For Medical Education And Research | Nanoparticle complexes of rituximab, albumin and pacltaxel |
US10376580B2 (en) | 2012-10-01 | 2019-08-13 | Mayo Foundation For Medical Education And Research | Methods of treating cancer with antibody-albumin nanoparticle complexes comprising albumin, trastuzumab, and paclitaxel |
US9757453B2 (en) | 2012-10-01 | 2017-09-12 | Mayo Foundation For Medical Education And Research | Nanoparticle complexes of anti-CD20 antibodies, albumin and paclitaxel |
US10406224B2 (en) | 2012-10-01 | 2019-09-10 | Mayo Foundation For Medical Education And Research | Nanoparticle complexes of paclitaxel, trastuzumab, and albumin |
US10413606B2 (en) | 2012-10-01 | 2019-09-17 | Mayo Foundation For Medical Education And Research | Methods for treating cancer with nanoparticle complexes of albumin-bound paclitaxel and anti-VEGF antibodies |
US10420839B2 (en) | 2012-10-01 | 2019-09-24 | Mayo Foundation For Medical Education And Research | Methods for treating CD52-expressing cancers using compositions comprising nanoparticle complexes of paclitaxel, alemtuzumab, and albumin |
US10441656B2 (en) | 2012-10-01 | 2019-10-15 | Mayo Foundation For Medical Education And Research | Nanoparticle complexes of paclitaxel, cetuximab, and albumin, and methods of making the same |
US10279036B2 (en) | 2012-10-01 | 2019-05-07 | Mayo Foundation For Medical Education And Research | Antibody-albumin nanoparticle complexes comprising albumin, bevacizumab, and paclitaxel, and methods of making and using the same |
US10478495B2 (en) | 2012-10-01 | 2019-11-19 | Mayo Foundation For Medical Education And Research | Methods for treating cancer using nanoparticle complexes of paclitaxel, cetuximab, and albumin |
US10493150B2 (en) | 2012-10-01 | 2019-12-03 | Mayo Foundation For Medical Education And Research | Nanoparticle complexes of paclitaxel, alemtuzumab, and albumin |
US10213513B2 (en) | 2014-06-16 | 2019-02-26 | Mayo Foundation For Medical Education And Research | Treating myelomas |
US11285221B2 (en) | 2014-06-16 | 2022-03-29 | Mayo Foundation For Medical Education And Research | Treating myelomas |
US10610484B2 (en) | 2014-10-06 | 2020-04-07 | Mayo Foundation For Medical Education And Research | Methods of using albumin-CD20 paclitaxel nanoparticle complex compositions for treating cancer |
US10322084B2 (en) | 2014-10-06 | 2019-06-18 | Mayo Foundation For Medical Education And Research | Carrier-antibody compositions and methods of making and using the same |
US10596112B2 (en) | 2014-10-06 | 2020-03-24 | Mayo Foundation For Medical Education And Research | Methods of using albumin-antibody nanoparticle complex compositions for treating cancer |
US10300016B2 (en) | 2014-10-06 | 2019-05-28 | Mayo Foundation For Medical Education And Research | Carrier-antibody compositions and methods of making and using the same |
US10624846B2 (en) | 2014-10-06 | 2020-04-21 | Mayo Foundation For Medical Education And Research | Lyophilized compositions comprising albumin-antibody paclitaxel nanoparticle complexes |
US11433023B2 (en) | 2014-10-06 | 2022-09-06 | Mayo Foundation For Medical Education And Research | Albumin-PD-1 paclitaxel nanoparticle complex compositions and methods of making and using the same |
US10391055B2 (en) | 2014-10-06 | 2019-08-27 | Mayo Foundation For Medical Education And Research | Carrier-antibody compositions and methods of making and using the same |
US10772833B2 (en) | 2014-10-06 | 2020-09-15 | Mayo Foundation For Medical Education And Research | Albumin-CTLA-4 paclitaxel nanopartilce complex compositions and methods of making and using the same |
US10780050B2 (en) | 2014-10-06 | 2020-09-22 | Mayo Foundation For Medical Education And Research | Lyophilized compositions comprosing albumin-EGFR paclitaxel nanoparticle complexes |
US10780049B2 (en) | 2014-10-06 | 2020-09-22 | Mayo Foundation For Medical Education And Research | Lyophilized compositions comprising albumin-rankl or albumin-GD2 paclitaxel nanoparticle complexes |
US10966923B2 (en) | 2014-10-06 | 2021-04-06 | Mayo Foundation For Medical Education And Research | Carrier-antibody compositions and methods of making and using the same |
US10993912B2 (en) | 2014-10-06 | 2021-05-04 | Mayo Foundation For Medical Education And Research | Carrier-antibody compositions and methods of making and using the same |
US10993911B2 (en) | 2014-10-06 | 2021-05-04 | Mayo Foundation For Medical Education And Research | Carrier-antibody compositions and methods of making and using the same |
US10596111B2 (en) | 2014-10-06 | 2020-03-24 | Mayo Foundation For Medical Education And Research | Methods of making lyophilized compositions comprising albumin-VEGF paclitaxel nanoparticle complexes |
US11241387B2 (en) | 2015-08-18 | 2022-02-08 | Mayo Foundation For Medical Education And Research | Carrier-binding agent compositions and methods of making and using the same |
US10561726B2 (en) | 2015-10-06 | 2020-02-18 | Vavotar Life Sciences LLC | Methods of treating cancer using compositions of antibodies and carrier proteins with antibody pretreatment |
US11660339B2 (en) | 2015-10-06 | 2023-05-30 | Mayo Foundation For Medical Education And Research | Methods of treating cancer using compositions of antibodies and carrier proteins with antibody pretreatment |
US11571469B2 (en) | 2016-01-07 | 2023-02-07 | Mayo Foundation For Medical Education And Research | Methods of treating cancer with interferon wherein the cancer cells are HLA negative or have reduced HLA expression |
US11351254B2 (en) | 2016-02-12 | 2022-06-07 | Mayo Foundation For Medical Education And Research | Hematologic cancer treatments |
US11305020B2 (en) | 2016-03-21 | 2022-04-19 | Mayo Foundation For Medical Education And Research | Methods for reducing toxicity of a chemotherapeutic drug |
US11878061B2 (en) | 2016-03-21 | 2024-01-23 | Mayo Foundation For Medical Education And Research | Methods for improving the therapeutic index for a chemotherapeutic drug |
US11655301B2 (en) | 2016-04-06 | 2023-05-23 | Mayo Foundation For Medical Education And Research | Carrier-binding agent compositions and methods of making and using the same |
US10618969B2 (en) | 2016-04-06 | 2020-04-14 | Mayo Foundation For Medical Education And Research | Carrier-binding agent compositions and methods of making and using the same |
US11160876B2 (en) | 2016-09-01 | 2021-11-02 | Mayo Foundation For Medical Education And Research | Methods and compositions for targeting t-cell cancers |
US11548946B2 (en) | 2016-09-01 | 2023-01-10 | Mayo Foundation For Medical Education And Research | Carrier-PD-L1 binding agent compositions for treating cancers |
US11311631B2 (en) | 2016-09-06 | 2022-04-26 | Mayo Foundation For Medical Education And Research | Paclitaxel-albumin-binding agent compositions and methods for using and making the same |
US11590098B2 (en) | 2016-09-06 | 2023-02-28 | Mayo Foundation For Medical Education And Research | Methods of treating triple-negative breast cancer using compositions of antibodies and carrier proteins |
US11872205B2 (en) | 2016-09-06 | 2024-01-16 | Mayo Foundation For Medical Education And Research | Methods of treating triple-negative breast cancer using compositions of antibodies and carrier proteins |
US11427637B2 (en) | 2016-09-06 | 2022-08-30 | Mayo Foundation For Medical Education And Research | Methods of treating PD-L1 expressing cancer |
Also Published As
Publication number | Publication date |
---|---|
US9387244B2 (en) | 2016-07-12 |
EP2310006A4 (en) | 2012-04-25 |
EP3266453A1 (en) | 2018-01-10 |
US20160304597A1 (en) | 2016-10-20 |
WO2010003057A2 (en) | 2010-01-07 |
US20140056909A1 (en) | 2014-02-27 |
WO2010003057A3 (en) | 2010-04-01 |
US10287344B2 (en) | 2019-05-14 |
EP2310006A2 (en) | 2011-04-20 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US10287344B2 (en) | Methods for reducing global chronic inflammation and the presence of melanoma | |
Prins et al. | Gene expression profile correlates with T-cell infiltration and relative survival in glioblastoma patients vaccinated with dendritic cell immunotherapy | |
von Euw et al. | CTLA4 blockade increases Th17 cells in patients with metastatic melanoma | |
Datta et al. | Anti-HER2 CD4+ T-helper type 1 response is a novel immune correlate to pathologic response following neoadjuvant therapy in HER2-positive breast cancer | |
Gray et al. | Phosphatidylserine-targeting antibodies augment the anti-tumorigenic activity of anti-PD-1 therapy by enhancing immune activation and downregulating pro-oncogenic factors induced by T-cell checkpoint inhibition in murine triple-negative breast cancers | |
Akasaki et al. | Phase I/II trial of combination of temozolomide chemotherapy and immunotherapy with fusions of dendritic and glioma cells in patients with glioblastoma | |
Bjoern et al. | Safety, immune and clinical responses in metastatic melanoma patients vaccinated with a long peptide derived from indoleamine 2, 3-dioxygenase in combination with ipilimumab | |
US20150335679A1 (en) | Methods and Compositions for Treating Malignancies with Dendritic Cells | |
Li et al. | Functional interleukin-17 receptor A are present in the thyroid gland in intractable Graves disease | |
Shevchenko et al. | Autologous dendritic cells and activated cytotoxic T‑cells as combination therapy for breast cancer | |
Botticelli et al. | The 5-Ws of immunotherapy in head and neck cancer | |
Rawat et al. | The CD200-CD200R cross-talk helps Leishmania donovani to down regulate macrophage and CD4+ CD44+ T cells effector functions in an NFκB independent manner | |
JP2015524252A (en) | Methods for determining personalized treatment compositions for prostate cancer and breast cancer | |
Hirschhorn-Cymerman et al. | T cell immunotherapies trigger neutrophil activation to eliminate tumor antigen escape variants | |
Liu et al. | Trichostatin A-modified vaccine provides superior protection against ovarian cancer formation and development | |
Srivastava | Therapeutic Intervention in Skin Cancer: Future Prospects | |
WO2022194401A1 (en) | Combination therapy for cancer | |
Lorrey et al. | Intracranial tumors elicit systemic sympathetic hyperactivity that limits immunotherapeutic responses | |
Dudzinski | Leptin Repolarizes Tumor-Associated Macrophages to Boost Immunotherapy Efficacy in Obesity | |
und Tumorimmunität | Steroid Hormones and Cancer Immunity–learning from Adrenocortical Carcinoma | |
Landwehr | Steroid Hormones and Cancer Immunity-learning from Adrenocortical Carcinoma | |
Parney et al. | Neuro-Oncology Advances | |
Turbitt Jr | The Effects of Changes in Energy Balance on Immune Regulation and Tumor Progression in the 4T1. 2 Mammary Tumor Model | |
Boi | Overcoming obesity-induced immunotherapeutic impairment | |
TW202317757A (en) | Modified t cells, pharmaceutical composition, manufacturing method thereof, and method of using the same |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AS | Assignment |
Owner name: MAYO FOUNDATION FOR MEDICAL EDUCATION AND RESEARCH Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:MARKOVIC, SVETOMIR N.;REEL/FRAME:029000/0242 Effective date: 20110111 |
|
STCB | Information on status: application discontinuation |
Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION |