CN104830769B - Butylated hydroxy anisole is to the application in central memory t cell amplification in vitro - Google Patents
Butylated hydroxy anisole is to the application in central memory t cell amplification in vitro Download PDFInfo
- Publication number
- CN104830769B CN104830769B CN201510228515.1A CN201510228515A CN104830769B CN 104830769 B CN104830769 B CN 104830769B CN 201510228515 A CN201510228515 A CN 201510228515A CN 104830769 B CN104830769 B CN 104830769B
- Authority
- CN
- China
- Prior art keywords
- cell
- central memory
- butylated hydroxy
- hydroxy anisole
- vitro
- 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.)
- Expired - Fee Related
Links
- 239000004255 Butylated hydroxyanisole Substances 0.000 title claims abstract description 39
- 235000019282 butylated hydroxyanisole Nutrition 0.000 title claims abstract description 39
- 210000003071 memory t lymphocyte Anatomy 0.000 title claims abstract description 19
- 229940043253 butylated hydroxyanisole Drugs 0.000 title claims abstract description 17
- 238000000338 in vitro Methods 0.000 title claims abstract description 16
- CZBZUDVBLSSABA-UHFFFAOYSA-N butylated hydroxyanisole Chemical compound COC1=CC=C(O)C(C(C)(C)C)=C1.COC1=CC=C(O)C=C1C(C)(C)C CZBZUDVBLSSABA-UHFFFAOYSA-N 0.000 title claims abstract 8
- 230000003321 amplification Effects 0.000 title description 9
- 238000003199 nucleic acid amplification method Methods 0.000 title description 9
- 210000001744 T-lymphocyte Anatomy 0.000 claims abstract description 47
- 238000000034 method Methods 0.000 claims abstract description 12
- 210000004027 cell Anatomy 0.000 claims description 34
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims description 9
- 210000004369 blood Anatomy 0.000 claims description 9
- 239000008280 blood Substances 0.000 claims description 9
- 238000010790 dilution Methods 0.000 claims description 6
- 239000012895 dilution Substances 0.000 claims description 6
- 239000012980 RPMI-1640 medium Substances 0.000 claims description 5
- 239000011324 bead Substances 0.000 claims description 5
- 210000004698 lymphocyte Anatomy 0.000 claims description 4
- 239000002609 medium Substances 0.000 claims description 4
- 238000000926 separation method Methods 0.000 claims description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 3
- 210000001616 monocyte Anatomy 0.000 claims description 3
- 229920006395 saturated elastomer Polymers 0.000 claims description 3
- 238000004113 cell culture Methods 0.000 abstract description 7
- 238000009169 immunotherapy Methods 0.000 abstract description 5
- 238000011161 development Methods 0.000 abstract description 3
- MRBKEAMVRSLQPH-UHFFFAOYSA-N 3-tert-butyl-4-hydroxyanisole Chemical compound COC1=CC=C(O)C(C(C)(C)C)=C1 MRBKEAMVRSLQPH-UHFFFAOYSA-N 0.000 description 10
- 230000000694 effects Effects 0.000 description 8
- 238000001514 detection method Methods 0.000 description 7
- 206010028980 Neoplasm Diseases 0.000 description 5
- 239000003963 antioxidant agent Substances 0.000 description 4
- 230000003078 antioxidant effect Effects 0.000 description 4
- 235000006708 antioxidants Nutrition 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 239000003814 drug Substances 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 102100036301 C-C chemokine receptor type 7 Human genes 0.000 description 2
- 101000716065 Homo sapiens C-C chemokine receptor type 7 Proteins 0.000 description 2
- 101001018097 Homo sapiens L-selectin Proteins 0.000 description 2
- 102100033467 L-selectin Human genes 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 230000011712 cell development Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000007619 statistical method Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 108010002586 Interleukin-7 Proteins 0.000 description 1
- SHGAZHPCJJPHSC-NUEINMDLSA-N Isotretinoin Chemical compound OC(=O)C=C(C)/C=C/C=C(C)C=CC1=C(C)CCCC1(C)C SHGAZHPCJJPHSC-NUEINMDLSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- RYMZZMVNJRMUDD-UHFFFAOYSA-N SJ000286063 Natural products C12C(OC(=O)C(C)(C)CC)CC(C)C=C2C=CC(C)C1CCC1CC(O)CC(=O)O1 RYMZZMVNJRMUDD-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 210000000612 antigen-presenting cell Anatomy 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000012636 effector Substances 0.000 description 1
- 238000013401 experimental design Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 229960005280 isotretinoin Drugs 0.000 description 1
- 230000002045 lasting effect Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000006386 memory function Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000009456 molecular mechanism Effects 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- RYMZZMVNJRMUDD-HGQWONQESA-N simvastatin Chemical compound C([C@H]1[C@@H](C)C=CC2=C[C@H](C)C[C@@H]([C@H]12)OC(=O)C(C)(C)CC)C[C@@H]1C[C@@H](O)CC(=O)O1 RYMZZMVNJRMUDD-HGQWONQESA-N 0.000 description 1
- 229960002855 simvastatin Drugs 0.000 description 1
- 229940126586 small molecule drug Drugs 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 238000011287 therapeutic dose Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
Landscapes
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides the application that butylated hydroxy anisole is expanded in vitro to central memory t cell, the concentration of the butylated hydroxy anisole is 5 ~ 40uM, and optimum concentration is 20uM, and the central memory t cell is CD4+T cell.Application of the butylated hydroxy anisole in vitro in T cell culture is made public for the first time in the present invention, method safety is effectively, with low cost, and good technical support is provided for the extensive development of adoptive immunotherapy.
Description
Technical field
The invention belongs to central memory t cell Amplification Technologies field, more particularly, to butylated hydroxy anisole
To the application in central memory t cell amplification in vitro.
Background technology
Butylated hydroxy anisole（English：Butylated hydroxyanisole, abbreviation BHA）, it is a kind of antioxidant,
For two kinds of isomer 2- tertiary butyl-4-hydroxies anisoles and the mixture of 3- tertiary butyl-4-hydroxy anisoles, BHA master
It is in food to want purposes（Including animal foodstuff）, packaging for foodstuff, play antioxidant and preservative in cosmetics and oil product
Effect.In addition, it is also used for the anti-oxidant of medicine, such as Accutane, Simvastatin.
Because T cell plays an important role in immune system, increasing research group is attempted with T cell
The mode for the treatment of of adopting carries out oncotherapy.Central memory t cell（TCM）Ability with self-renewing, and response
Time is short, long action time, to the Small side effects of human body.This year, multiple seminar's reports find the T with memory function
After cell is fed back in tumor patient body, therapeutic effect is substantially and the continued treatment time is long, it is possible to reduce the pain of patient with
And the excessive bio-safety risk of reduction cell injuring model number of times.But relative populations are less in human body, and due to external
The T cell of culture can make T cell lose vigor for a long time, and most cell differentiations are the effector cell of terminal differentiation so that
Feed back the internal T cell time-to-live too short, it is impossible to produce the function of lasting killing tumor cell, therefore how to obtain in vitro
Obtain the new study hotspot that substantial amounts of memory t cell is cellular immunotherapy.
At present, external many research groups are applied to the external of T cell by building artificial antigen presenting cells
Culture, improves the time-to-live of T cell, for adoptive immunotherapy, but to obtain the T cell of sufficient amount, and technical requirements are complicated,
The success rate of culture is relatively low.When carrying out T cell treatment, main policies are to first pass through clonal expansion, and obtain sufficient amount has
The T cell of killing ability, can be co-cultured with the presenting cells such as DC, can also be directly with associated tumor antigen stimulation cell clone
Amplification, is obtained with the specific T cell of particular tumor antigens.The T cell that such a method culture is obtained turns base without any
Because of modification, security is higher.But the cell of this tumour high response needs to be expanded to the 10 of clinical needs9-1011Therapeutic dose
Quantity number, process duration is long, and the time, which is also one, for tumor patient needs the factor of more considerations.And
And the time of in vitro culture is long, cell is easily ageing, and vigor declines, mostly the cell of terminal differentiation, there is preferable effect in vitro
Really, but in vivo the time-to-live is shorter, and therapeutic effect is impacted.
The content of the invention
The technical problem to be solved in the present invention is to overcome the shortcomings of existing central memory t cell Amplification Technologies, is carried
For butylated hydroxy anisole to the application in central memory t cell amplification in vitro.
Above-mentioned purpose of the present invention is achieved through the following technical solutions：
The invention provides the application that butylated hydroxy anisole is expanded in vitro to central memory t cell.
Preferably, the concentration of the butylated hydroxy anisole is 5 ~ 40uM.
It is highly preferred that the concentration of the butylated hydroxy anisole is 20uM.
Preferably, the central memory t cell is CD4+T cell.
Preferably, the extracting method of the central memory t cell is first by periphery component blood with containing 2% BSA and 0.5%
EDTA PBS presses 1:4 are well mixed；Then blood after dilution be slowly added to the top of lymphocyte separation medium, the two
Ratio is 1:1；Centrifuge the min of 300 × g 30；It is careful to draw monocyte, insert in another centrifuge tube, add containing for 5 times of volumes
0.5% BSA and 2% EDTA PBS dilutions, after being well mixed, the min of 300 × g 15；Repeat previous step once；It is incubated CD4
+ T cell the moon selects primary antibody, 15 min；Diluted with the PBS containing 0.5% BSA and 2% EDTA of 10 times of volumes, after being well mixed,
300×g 12 min；It is incubated the secondary antibody with magnetic bead, 30 min；Cross post and carry out cell sorting, obtain CD4+ T cells.
Preferably, the cultural method of the central memory t cell is in 5% CO2, will under the conditions of saturated humidity and 37 DEG C
5×105The CD4 in individual/hole+T cell is placed in RPMI1640 of 1 mL containing 10% hyclone and cultivated.
Compared with prior art, the present invention has advantages below and beneficial effect：
Present invention firstly discloses the application in butylated hydroxy anisole in vitro T cell culture, method safety is effective, into
This is cheap, and good technical support is provided for the extensive development of adoptive immunotherapy.
Brief description of the drawings
Fig. 1 is to CD4 when BHA concentration is 20uM+CD4 in T cell+The detection of TCM proportions.
Fig. 2 is the BHA of various concentrations to central Memorability CD4+The ratio expanding effect of T cell.
Embodiment
The present invention is further illustrated below in conjunction with specific embodiment, but embodiment does not do any type of to the present invention
Limit.Unless stated otherwise, the reagent of the invention used, method and apparatus is the art conventional reagent, methods and apparatus.
Unless stated otherwise, agents useful for same and material of the present invention are purchased in market.
Embodiment 1
1st, test material prepares
The periphery component blood of people is provided by Guangzhou Blood Center, and the blood sample of 24 parts of normal persons has been randomly selected in experiment
This.
Main agents：Hyclone is purchased from GIBCO companies；RPMI1640 culture mediums are purchased from INVERTROGEN companies；
BHA is purchased from INVERTROGEN companies；Flow cytomery antibody anti-CD45RA-Texas Red, anti-CCR7-
AF700 anti-CD62L-PE-cy7 be purchased from BD companies；Lymphocyte separation medium is limited purchased from the positive biological products of Tianjin Hao
Responsible company；CD4+T cell Solid phase magnetic bead is purchased from BD companies.
Main laboratory apparatus：Table model high speed centrifuge（Eppendorf Centrifuge 5810R）, flow cytometer
（Beckman Coulter companies）、CO2Cell culture incubator（Thermo SCIENTIFIC）, Biohazard Safety Equipment（Thermo
SCIENTIFIC）, inverted biologic microscope（Leica）, magnetic bead sorting magnetic frame（BD Pharmigen）.
Cell culture
2.1 cell extractions are separated
By periphery component blood and PBS（Containing 2% BSA, 0.5% EDTA）By 1:4 are well mixed；Then after diluting
Blood be slowly added to the top of lymphocyte separation medium, the ratio of the two is 1:1；Centrifuge the min of 300 × g 30；It is careful to draw
Monocyte, is inserted in another centrifuge tube, adds the PBS of 5 times of volumes（Containing 0.5% BSA, 2% EDTA）Dilution, is well mixed
Afterwards, the min of 300 × g 15；Repeat previous step once；It is incubated CD4+ T cell the moon and selects primary antibody, 15 min；The PBS of 10 times of volumes
（Containing 0.5% BSA, 2% EDTA）Dilution, after being well mixed, the min of 300 × g 12；It is incubated the secondary antibody with magnetic bead, 30 min；
Cross post and carry out cell sorting, obtain CD4+ T cells, add the RPMI1640 containing 10% hyclone and cell is resuspended, count cell
Cell is adjusted afterwards to required concentration.
2.2 condition of culture
In 5% CO2, saturated humidity and 37oUnder C, by 5 × 105The CD4 in individual/hole+T cell is placed in 1 mL containing 10% tire ox blood
Cultivated in clear RPMI1640.
Plating cells and agent-feeding treatment
3.1 bed board
By the CD4 sorted out+T cell is layered in 12 orifice plates, according to experimental design, be 5 per cell number expected from hole ×
105。
3.2 agent-feeding treatment
To the CD4 after bed board+T cell carries out 5 kinds of different processing, and every kind of processing sets three multiple holes as parallel right
According to, and at the 4th day of cell culture, the 7th day, the 10th day, apply within the 13rd day corresponding stimulate.
Cell subsets analysis and the detection of cell quantity
Cell culture, after 15 days, different donors are drawn respectively（n=24）Culture hole in cell, use cell counter
Counted；5 × 10 are drawn in the corresponding culture hole of slave phase5Individual cell, the min of 300 × g 15, cell is washed with PBS twice,
It is incubated detection CD4+TCM streaming antibody（CD45RA、CCR7、CD62L）Half an hour；Wash streaming antibody off, add PBS dilute
Respective concentration is released, with flow cytomery.Control group and the CD4 that experimental group is same donor+ T cells, Mei Geshi
Between put and take the cell of different donors to be detected respectively.
Statistics software and statistical method
Statistical analysis is carried out using SPSS13.0 analysis softwares.All results of measurement data use mean ± standard deviation table
Show.Experimental group is compared with control group to be examined using t, P<0.01 is to have significant difference.
Experimental result
Flow cytometer detections of 6.1 BHA to CD4+TCM proportions in CD4+T cells
In order to determine the original ratio for adding TCM in the CD4+ T cells that BHA is stimulated, by CD4+T cells are with 5 × 106
Individual/hole is laid in 24 orifice plates, adds anti-CD3 antibody, anti-CD28 antibody, IL-7 and IL-15.Control group and experimental group
Streaming antibody labeled cells are used in 15 d, are then detected sample by flow cytometer.Negative from CD45RA
In cell, it is the CD4 to be observed to choose the double positive cells of CD62L and CCR7+TCM.According to above by flow cytometer
Sort CD4+TCM method, to CD4+TCM is in CD4+Ratio in T cell detected, as shown in Figure 1.
The BHA of 6.2 various concentrations is for CD4+ The detection of T cell expanding effect
The time point that selection BHA handles the 15th day is detected, is added without the CD4 of BHA control group+TCM cells are accounted for
Total CD4+The ratio of T cell is 16.06%；BHA concentration is that 5uM ratios are 22.59%；The ratio that BHA concentration is 10uM is
25.91%；The ratio that BHA concentration is 20uM is 29.09%；The ratio that BHA concentration is 40uM is 28.33%.Test result indicates that,
The BHA of various concentrations is for CD4+There is dose-dependent trend in the amplification effect of T cell, such as in the case of less than 20uM
Shown in Fig. 2.
Interpretation of result
We promote CD4 using small-molecule drug BHA first+The amplification of TCM cells in vitro.Experimental group is compared with control group
Compared to CD4+The ratio of TCM cells is in CD4+Have in T cell and significantly raise.
Among the immune treatment of adopting of T cell, if substantial amounts of adoptive transfer T cell can produce radical response, in experiment
During, we be have found in the case where T cell sum is constant, and substantial amounts of T cells are converted into CD4+TCM cells
Method.We, which also further have found BHA, in experiment can promote CD4+The optimum concentration that TCM is expanded in vitro：20uM.I
Find, using relatively low-dose BHA handle cell after, CD4+ TCM cells are in CD4+Lift exists aobvious in ratio in T cell
The dose-dependant trend of work；And lift effect when BHA concentration reaches 20uM, in ratio and substantially tend towards stability（Fig. 2）, this may
It is due to that drug concentration is excessive so that the growing environment of cell there occurs larger change and have impact on the normal growth of cell.
In this experiment, counted after choose BHA processing cells 15 days and detection is due to the CD4 in peripheral blood+TCM's rises
Beginning quantity is considerably less, CD4+T cell needs longer time could be under anti-CD3 and anti-CD28 Co stituations gradually
Form more CD4+TCM.Experimental result also indicates that selection culture long period rear carries out detection and can obtain more preferable effect
（Fig. 2）.
BHA is a kind of conventional food and medicine antioxidant.BHA T cell cultures in vitro are elaborated in the present invention first
In brand-new application, provide good technical support for the extensive development of adoptive immunotherapy, its molecular mechanism needs subsequently
Further research.
Claims (6)
1. the application that butylated hydroxy anisole is expanded in vitro to central memory t cell.
2. application according to claim 1, it is characterised in that the concentration of the butylated hydroxy anisole is 5 ~ 40 μM.
3. application according to claim 2, it is characterised in that the concentration of the butylated hydroxy anisole is 20 μM.
4. the application according to claims 1 to 3 any one, it is characterised in that the central memory t cell is CD4+
T cell.
5. application according to claim 1, it is characterised in that the extracting method of the central memory t cell is first will
Periphery component blood presses 1 with the PBS containing 2% BSA and 0.5% EDTA:4 are well mixed；Then the blood after dilution is slowly added to
The top of lymphocyte separation medium, the ratio of the two is 1:1；Centrifuge the min of 300 × g 30；It is careful to draw monocyte, insert
In another centrifuge tube, the dilutions of the PBS containing 0.5% BSA and 2% EDTA of 5 times of volumes are added, after being well mixed, 300 × g are centrifuged
15 min；Repeat previous step once；It is incubated CD4+T cell the moon selects primary antibody, 15 min；Contain 0.5% BSA with 10 times of volumes
Diluted with 2% EDTA PBS, after being well mixed, centrifuge the min of 300 × g 12；It is incubated the secondary antibody with magnetic bead, 30 min；Cross
Post carries out cell sorting, obtains CD4+T cell.
6. the application that butylated hydroxy anisole according to claim 1 is expanded in vitro to central memory t cell, its
It is characterised by, the cultural method of the central memory t cell is in 5% CO2, under the conditions of saturated humidity and 37 DEG C by 5 × 105
The CD4 in individual/hole+T cell is placed in RPMI1640 of 1 mL containing 10% hyclone and cultivated.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510228515.1A CN104830769B (en) | 2015-05-07 | 2015-05-07 | Butylated hydroxy anisole is to the application in central memory t cell amplification in vitro |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510228515.1A CN104830769B (en) | 2015-05-07 | 2015-05-07 | Butylated hydroxy anisole is to the application in central memory t cell amplification in vitro |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104830769A CN104830769A (en) | 2015-08-12 |
| CN104830769B true CN104830769B (en) | 2017-08-04 |
Family
ID=53808971
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510228515.1A Expired - Fee Related CN104830769B (en) | 2015-05-07 | 2015-05-07 | Butylated hydroxy anisole is to the application in central memory t cell amplification in vitro |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104830769B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103379921A (en) * | 2010-12-17 | 2013-10-30 | 人类起源公司 | Treatment of spinal cord injury and traumatic brain injury using amnion derived adherent cells |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040203142A1 (en) * | 2003-04-14 | 2004-10-14 | Reliance Life Sciences Pvt. Ltd. | Growth of neural precursor cells using umbilical cord blood serum and a process for the preparation thereof for therapeutic purposes |
-
2015
- 2015-05-07 CN CN201510228515.1A patent/CN104830769B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103379921A (en) * | 2010-12-17 | 2013-10-30 | 人类起源公司 | Treatment of spinal cord injury and traumatic brain injury using amnion derived adherent cells |
Non-Patent Citations (5)
| Title |
|---|
| Cheryl E.Rockwell等.Th2 Skewing by Activation of Nrf2 in CD4+ T Cells.《The Journal of Immunology》.2012,第188卷第1630-1637页. * |
| Fang-Ming Hung等.Butylated hydroxyanisole affects immunomodulation and promotes macrophage phagocytosis in normal BALB/c mice.《Molecular Medicine Reports》.2012,第5卷第683-687页. * |
| John Whysner等.Butylated Hydroxyanisole Mechanistic Data and Risk Assessment:Conditional Species-Specific Cytotoxicity,Enhanced Cell Proliferation,and Tumor Promotion.《Pharmacology & Therapeutics》.1996,第71卷(第1-2期),第137-151页. * |
| P.A.E.L.Schiderman等.Induction of oxidative DNA damage and enhancement of cell proliferation in human lymphocytes in vitro by butylated hydroxyanisole.《 Carcinogenesis 》.1995,第16卷(第3期),第507-512页. * |
| 王卫民等.BHA诱导人脐带间充质干细胞分化为神经细胞.《东南大学学报（医学版）》.2013,第32卷(第2期),第201-205页. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104830769A (en) | 2015-08-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107083360B (en) | A kind of method of external evoked amplification human antigen's non-specificity regulatory T cells | |
| CN103756963B (en) | A kind of method of amplification in vitro NK cell | |
| CN103756961B (en) | A kind of method of external evoked amplification NKT cell | |
| CN104894065B (en) | A kind of cultural method of NK cell culture mediums and NK cells | |
| US11278569B2 (en) | Natural killer cell containing exogenous mitochondrium and pharmaceutical composition comprising same | |
| CN107475192A (en) | It is a kind of to do lymphocyte populations and its external Efficient amplification method of the T cell as main component to remember | |
| Barrie et al. | Prostaglandin E2 and IL‐23 plus IL‐1β Differentially Regulate the Th1/Th17 Immune Response of Human CD161+ CD4+ Memory T Cells | |
| CN108060129A (en) | Regulatory T cells amplification in vitro method | |
| CN106566806A (en) | Method for in-vitro culture and enrichment of CD8+ T cells | |
| CN106661551A (en) | A method of generating multilineage potential cells from lymphocytes | |
| CN107603948B (en) | A method of direct external evoked human peripheral blood single nucleus cell becomes compound antigen-non-specific regulatory T cells | |
| CN106554942A (en) | A kind of efficient clinical grade CD56+The preparation method of group's immunocyte | |
| TWI224137B (en) | A process for proliferating natural killer cell, method for determining cytotoxic activity same, and therapeutic agent of tumor | |
| Yeap et al. | Immunomodulatory effect of Rhaphidophora korthalsii on natural killer cell cytotoxicity | |
| CN105106237B (en) | A kind of Efficient killing effect tumour cell biological agent | |
| CN101914497B (en) | Clinical N-CIK cell culture and quality control and identification kit and application | |
| CN104830769B (en) | Butylated hydroxy anisole is to the application in central memory t cell amplification in vitro | |
| US20220168348A1 (en) | Method for culturing allogeneic immune cell, immune cell culture obtained thereby, and immune cell therapeutic agent comprising same | |
| CN108441481A (en) | A kind of Chimeric antigen receptor T cell and its cultural method | |
| CN102925411A (en) | Application of vitamin C in in-vitro expansion of number of effector memory T cells | |
| CN110337446A (en) | CCR2 in adoptive cellular therapy+The t cell activation that candidate stem cell mediates | |
| CN104830767B (en) | Cefonicid sodium is to the application in central memory t cell amplification in vitro | |
| CN106479973B (en) | A kind of external IAK immunocyte cultural method | |
| CN104818250B (en) | Bisacody is to the application in central memory t cell amplification in vitro | |
| CN104830770B (en) | Ampicillin is to the application in central memory t cell amplification in vitro |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| EXSB | Decision made by sipo to initiate substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20170804 |