CN1320923C - 含有基于癌抑制基因wt1的产物的癌抗原和阳离子脂质体的癌疫苗 - Google Patents
含有基于癌抑制基因wt1的产物的癌抗原和阳离子脂质体的癌疫苗 Download PDFInfo
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Abstract
含有以癌抑制基因WT1的产物或其部分肽或修饰产物为活性成分的癌抗原和阳离子脂质体构成的癌疫苗。
Description
技术领域
本发明涉及到含有源于Wilms肿瘤的癌抑制基因WT1的产物的癌抗原和脂质转染试剂Lipofectin的癌疫苗。该癌疫苗作为针对以下癌症以及表达WT1的所有癌的抗癌疫苗是有用的,这些癌症包括白血病、骨髓异形成综合征、多发性骨髓瘤、恶性淋巴瘤等血液癌、或实体癌、例如胃癌、结肠癌、肺癌、乳腺癌、胚细胞癌、肝癌、皮肤癌、膀胱癌、***癌、子宫癌、子***、卵巢癌等。
背景技术
在用于排除异物的免疫***中,一般存在着体液免疫和细胞免疫,体液免疫涉及到作为抗原呈递细胞起着识别抗原作用的巨噬细胞、识别该巨噬细胞的抗原呈递产生各种淋巴因子,对其他T-细胞等进行活化的辅助性T细胞、由于该淋巴因子的作用向产生抗体细胞分化的B-淋巴细胞等;而细胞免疫涉及到接受抗原的呈递之后进行分化的杀伤性T-细胞攻击靶细胞并进行破坏。
目前,人们认为癌的免疫主要是由有杀伤性T-细胞参与的细胞免疫引起的。在通过杀伤性T-细胞引起的癌免疫中,对以主要组织相容性抗原(Major Histocompatibility Complex;MHC)I类和癌抗原的复合物的形式呈递的癌抗原进行识别后的前体T-细胞分化增殖,生成的杀伤性T-细胞攻击癌细胞,并进行破坏。此时,癌细胞在该细胞表面呈递MHCI类抗原和癌抗原的复合物,该复合物成为杀伤性T-细胞的靶子(Cur.Opin,Immunol.,5,709,1993;Cur.Opin,Immunol.,5,719,1993;Cell,82,13,1995;Immunol.Rev,146,167,1995)。
有人认为作为靶细胞的癌细胞上通过MHC1类抗原呈递的上述癌抗原是由癌细胞内合成的抗原蛋白质经细胞内蛋白酶加工生成的约8~12个氨基酸构成的肽(Cur.Opin,Immunol.,5,709,1993;Cur.Opin,Immunol.,5,719,1993;Cell,82,13,1995;Immunol.Rev,146,167,1995)。
Wilms肿瘤的癌抑制基因WT1(WT1基因)是根据对合并有Wilms肿瘤、无虹膜、泌尿生殖器异常、精神发育迟滞等WAGR综合征的分析作为Wilms肿瘤的原因基因之一从染色体11p13分离出来的基因(Gessler,M.等人、Nature,Vol.343,p.774-778,1990),基因组DNA约50kb,由10个外显子构成,其cDNA约3kb。由cDNA推定的氨基酸序列如序列1所示(Mol.Cell.Biol.,11,1707,1991)。
WT1基因在人白血病中高表达,如果用WT1反义寡聚物对白血病细胞进行处理,由该细胞的增殖被抑制(特开平9-104627号公报)等现象暗示WT1基因起着促进白血病细胞增殖的作用。另外WT1基因在胃癌、结肠癌、肺癌、乳腺癌、胚细胞癌、肝癌、皮肤癌、膀胱癌、***癌、子宫癌、子***、卵巢癌等实体癌中也高表达(特原平9-191635),判明WT1基因是白血病以及实体癌中的新的肿瘤标记。
由于这一标记,期望给药含有由WT1基因的表达产物的部分构成的约8~12个氨基酸的肽,对上述很大范围的癌能够作为癌疫苗发挥作用。然而,将那样的肽原封不动地直接给药很难起到癌疫苗的作用。预料其原因是由于给药的肽不能有效地送到抗原呈递细胞上的主要组织抗原类I。
作为阳离子脂质体的脂质转染试剂Lipofectin是人工脂N-[1-(2,3-二油酰氧)丙基]-N,N,N-氯化三甲铵和作为磷脂的二油酰磷脂酰乙醇胺的1∶1混合物,以前作为非病毒性的基因导入用载体引人注目,后来由于起到将肽抗原送到抗原呈递细胞表面的主要组织抗原I类的作用而正被人关注(临床免疫Vol.34,No.6,p842-847,2000)。然而,阳离子脂质体作为抗原载体具有的普遍性程度如何还不清楚,是否能够起到作为由癌抑制基因WT1的表达产物片段构成的癌抗原的载体的功能也不清楚。
发明的公开
因此,本发明的目的是提供含有来自WT1基因表达产物的癌抗原肽和作为该肽载体的有效物质构成的新的癌疫苗。
本发明人为了解决上述课题进行了各种研究,结果确认,由其中至少含有一个在WT1基因表达产物的氨基酸序列中预测在与小鼠和人的MHCI类以及MHCII类的结合中起着锚定氨基酸功能的氨基酸的连续的7~30个氨基酸构成的多肽具有癌抗原的功能,脂质转染试剂Lipofectin等阳离子脂质体可以作为该癌抗原的载体,完成了本发明。
因此,本发明提供含有鼠WT1表达产物或其部分的癌抗原以及阳离子脂质体的癌疫苗。在优选的方案中,本发明提供由其中含有从在对应于WT1的cDNA的序列1所示的氨基酸序列中推定起着用于与MHC抗原结合的锚定氨基酸作用的Phe,Tyr,Leu,Met,Asn以及Ile中选择出的至少一个氨基酸的6~30个氨基酸构成的肽作为活性成分的癌抗原和阳离子脂质体的癌疫苗。
另外本发明提供由其中含有从在对应于WT1的cDNA的序列2所示的氨基酸序列中推定起着用于与MHC抗原结合的锚定氨基酸作用的Met,Ler以及Val中选择出的至少一个氨基酸的7~30个氨基酸构成的肽作为活性成分的癌抗原和阳离子脂质体的癌疫苗。
附图的简单说明
图1A是表示在实施例1中,使用(3)RNAS细胞以及用(4)癌抗原肽Db126进行刺激的RNAS细胞,对癌抗原肽Db126和脂质转染试剂Lipofectin(LPF)的混合物(黑圆点)、用Db126进行脉冲后的脂多糖-胚细胞(脂多糖-blast)(白正方形)、单独脂质转染试剂Lipofectin(白三角)以及单独癌抗原肽Db126(白圈)的细胞毒性T细胞诱导能力进行比较的图,图B是表示使用(1)C1498细胞以及(2)导入了WT1基因的C1498细胞进行与上述A同样的实验后的结果图。图A表明癌抗原肽Db126和脂质转染试剂Lipofectin的组合具有诱导细胞毒性T细胞能力,B表示其活性是WT1特异的。
图2A是表示在实施例2中作为佐剂(载体)的脂质转染试剂Lipofectin对使用导入了WT1基因的C1498细胞的肽Db126的抗癌效果的效果图,图2B是表示使用C1498细胞进行同样实验的结果的图。表示被检测物质的记号与图1相同。A表示脂质转染试剂Lipofectin作为癌抗原肽Db126的佐剂(载体)是有效的,比较A和B表明抗癌效果是WT1特异的。
发明的实施形态
在本发明中,作为设计癌抗原肽时的基础,对小鼠MHC I类的Kb和Db以及人HLA的A0201进行选择,选择了预测与他们有高亲和性的肽。
根据Immunogenetics Vol.41,p.178-228(1995)的报道,作为与Kb结合的锚定氨基酸推测是第5位的Phe以及Try以及第8位的Leu以及Met等,而作为与Db结合的锚定氨基酸推测是第5位的Asn以及第9位的Met和Ile等。
另外,已知在癌细胞的表面通过MHC I类呈递的癌抗原肽的大小大约为8~12个氨基酸。因此,本发明的癌抗原肽是由至少含有序列1所示WT1基因产物的氨基酸序列中的Phe,Tyr,Leu,Met,Asn以及Ile中的一个氨基酸的7~30个氨基酸构成的肽。氨基酸的数目优选是8~12个,例如8或9个。
在本发明中,作为该肽的具体例子,如与MHC I类的Kb结合的肽,可以是由8个氨基酸构成的下列肽:
Kb45 Gly Ala Ser Ala
Tyr Gly Ser
Leu(序列3)
Kb330 Cys Asn Lys Arg
Tyr Phe Lys
Leu(序列4)
作为与MHC I类的Db结合的肽,例如可以是由9个氨基酸构成的下列肽:
Db126 Arg Met Phe Pro
Asn Ala Pro Tyr
Leu(序列5)
Db221 Tyr Ser Ser Asp
Asn Leu Tyr Gln
Met(序列6)
Db235 Cys Met Thr Trp
Asn Gln Met Asn
Leu(序列7)
在上述序列中加了下划线的氨基酸是推测起着锚定氨基酸作用的氨基酸。
无论哪一个肽与Kb或Db都有强~中等程度的结合亲和性(kd值),但将表现出最高亲和活性的Db126肽用于以后的实验中。
另外关于人,根据Immunogenetics Vol.41,p.178-228(1995)的报道,作为与人的HLA-A0201结合的锚定氨基酸,推测是从N-末端开始的第2位的Leu和Met,以及从N-末端开始的第9位Val和Leu。因此,这里列举了由在人WT1蛋白质的氨基酸序列(Mol.Coll..Biol.Vol.11,p.1707-1712,1991)(序列2)中,符合上述条件的9个氨基酸构成的下列2种肽。
Db126:Arg
Met Phe Pro Asn Ala Pro Tyr
Leu(序列5)
(与小鼠中的Db126序列相同)
WH 187:Ser
Leu Gly Glu Gln Gln Tyr Ser
Val(序列8)
(下划线表示锚定氨基酸)
另外本发明的癌抗原肽也可以是在作为WT1基因表达产物的部分的肽中导入了氨基酸置换等修饰的肽。作为这样的被修饰的肽的例子如含有以下氨基酸序列:Cys Tyr Thr Trp Asn Gln Met Asn Leu(序列9),由9~30个氨基酸构成的肽作为活性成分的癌抗原肽。具体的例子如含有上述的肽Db235(序列7)的2位Met变换为Tyr的氨基酸序列:Cys Tyr ThrTrp Asn Gln Met Asn Leu(序列9)的肽。
作为阳离子脂质体,可列举例如,含有N-[1-(2,3-二油酰氧)丙基]-N,N,N-氯化三甲铵(DOTMA)、N-[1-(2,3-二油酰氧)丙基]-N,N,N-三甲铵硫酸甲酯(DOTAP)、或双十八烷基酰胺甘氨酰精胺(DOGS)或他们与中性脂的混合物的脂质体。
作为中性脂,可列举例如,卵磷脂、溶血卵磷脂、[神经]鞘髓磷脂、磷脂酸、磷脂酸乙醇胺、二油酰磷脂酰乙醇胺(DOPE)等。作为混合物的例子,可列举例如,作为人工脂的N-[1-(2,3-二油酰氧)丙基]-N,N,N-氯化三甲铵(DOTMA)和作为磷脂的二油酰磷脂酰乙醇胺(DOPE)的1∶1混合物的脂质转染试剂Lipofectin。
本发明的癌疫苗可以用于伴随着WT1基因表达水平上升的癌,例如白血病、骨髓异形成综合征、多发性骨髓瘤、恶性淋巴瘤等血液癌、胃癌、结肠癌、肺癌、乳腺癌、胚细胞癌、肝癌、皮肤癌、膀胱癌、***癌、子宫癌、子***、卵巢癌等实体癌的预防和治疗。该疫苗可以通过口服给药、非口服给药、例如腹腔内给药、皮下给药、皮内给药、肌肉内给药、静脉内给药、鼻腔内给药等进行给药。
本发明的癌疫苗的给药量一般是每天0.1μ~1mg/kg。
实施例
以下通过实施例阐明本发明的癌疫苗的有用性。
实施例1
脂多糖-胚细胞(LPS-blast)的制备
从C57BL/6鼠中回收脾细胞,将该脾细胞于含有脂多糖(LPS)(10μg/mL)的完全RPMI培养基中培养3天。将该细胞洗净后,于含有癌抗原肽Db126(1μM)以及卵清清蛋白(OVA)(100μg/mL)的完全RPMI培养基中培养2小时。将细胞洗净后,悬浮于2mL的HBSS(Hanks’balanced salt solution)中,该悬浮液作为脂多糖-胚细胞(LPS-blast)。
细胞毒性T细胞(CTL)诱导能力的评价
每隔一周向C57BL/6鼠的背部皮下注射共3次癌抗原肽Db126和脂质转染试剂Lipofectin(LPF)的混合物(Db126和LPF以1∶2的重量比混合后的混合物)(每只鼠10nmol的Db126),或作为阳性对照每隔一周向腹腔注射共3次的脂多糖-胚细胞(LPS-blast)(每只鼠1mL),进行免疫。在最终免疫10天后,回收脾细胞,作为效应细胞(effector)。用HBSS将经癌抗原肽Db126刺激(1μM、2小时、37℃、5%CO2条件下)的脾细胞洗净,得到刺激(stimulator)细胞。
将上述的效应细胞(5×106细胞/孔)和上述刺激细胞(2.5×106细胞/孔)混合,通过进行淋巴细胞·淋巴细胞混合培养,进行细胞毒性T细胞的体外2次抗原刺激。5天后,回收细胞毒性T细胞。将都用Na2 51CrO4标记的(0.56MBq/106cells、37℃、5%CO2条件下处理1小时)(1)C1498细胞、(2)导入WT1基因后使其表达的C1498细胞(C1498muWT1)、(3)RMA-S细胞以及(4)经癌抗原肽Db126刺激(1μM、5%CO2条件下处理1小时)的RMA-S细胞(Db126-pulsed RMA-S细胞)作为靶细胞,铺在96孔微量反应板中(104细胞/孔),然后向培养板中接种上述那样制备的细胞毒性T细胞。加入效应细胞,培养4小时,测定上清中游离的51Cr的放射活性。细胞损伤活性根据下式算出。
结果
首先,为了研究诱导的细胞毒性T细胞是否对癌抗原肽Db126特异,使用作为靶细胞的上述(3)RMA-S细胞和(4)Db126-pulsed RMA-S细胞,确认了癌抗原肽Db126特异性地诱导细胞毒性T细胞(图1.A)。另外为了研究被诱导的细胞毒性T细胞是否特异损伤WT1表达细胞,作为靶细胞使用上述(1)C1498细胞、(2)导入WT1基因的细胞C1498muWT1,确认了WT1特异的CTL的诱导(图1.B)。
实施例2.作为癌疫苗载体使用脂质转染试剂Lipofectin(LPF)时的
癌抗原特异的抗肿瘤效果
在实施例1中,由于通过使用作为癌抗原肽Db126的佐剂脂质转染试剂Lipofectin(LPF)表现出有效地诱导细胞毒性T细胞,为了进一步确认作为癌疫苗用佐剂的脂质转染试剂Lipofectin(LPF)的有用性,就作为佐剂(载体)使用脂质转染试剂Lipofectin进行免疫时的癌抗原特异的抗肿瘤效果进行了研究。
作为肿瘤模型使用导入了WT1基因的C1498细胞(C1498muWT1),作为免疫动物使用C57BL/6小鼠,而作为模型癌抗原使用肽Db126。即,象实施例1那样,每隔一周向C57BL/6小鼠的背部皮下注射共3次肽Db126和脂质转染试剂Lipofectin(LPF)的混合物(每只鼠10nmol),或每隔一周向C57BL/6小鼠的腹腔注射共3次脂多糖-胚细胞(LPS-blast)(每只鼠1mL),进行免疫。最终免疫一周后,将C1498muWT1细胞、或C1498细胞以2×106细胞/100mL的量移植到腹腔内。肿瘤疫苗效果通过测定随着时间推移的肿瘤直径,用以下式子算出肿瘤的大小进行评价。
[肿瘤的大小]=[(长径)×(短径)2]1/3
各组中肿瘤的大小达到20mm时,终止实验。
结果
作为癌疫苗用佐剂(载体)的脂质转染试剂Lipofectin(LPF)的评价,是以作为模型肿瘤抗原使用WT1,作为模型肿瘤使用导入了WT1基因的细胞(C1498muWT1细胞),进行Db126/脂质转染试剂Lipofectin(LPF)混合物免疫时对C1498muWT1细胞的抵抗性为指标进行的。结果,如果免疫肽Db126/脂质转染试剂Lipofectin(LPF)混合物,可以看到8例中有3例完全拒绝(图2.A)。
另外为了确认该抗肿瘤效果是WT1特异的,使用不表达WT1的C1498细胞,进行同样的研究,在免疫(a)肽Db126/脂质转染试剂Lipofectin(LPF)混合物、(b)游离的肽Db126以及(c)脂多糖-blast(LPS-blast)的任一种时,看不到与没有进行免疫的组的差别(图2.B)。因此,确认上述的抗肿瘤效果是WT1特异的。
序列表
<110>中外制药株式会社
真弓,忠范
杉山,治夫
<120>含有基于癌抑制基因WT1的产物的癌抗原和阳离子脂质体的癌疫苗
<130>K807
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Ser Leu Gly Gly Gly Gly Gly Cys Ala Leu Pro Val Ser Gly Ala Ala
20 25 30
Gln Trp Ala Pro Val Leu Asp Phe Ala Pro Pro Gly Ala Ser Ala Tyr
35 40 45
Gly Ser Leu Gly Gly Pro Ala Pro Pro Pro Ala Pro Pro Pro Pro Pro
50 55 60
Pro Pro Pro Pro His Ser Phe Ile Lys Gln Glu Pro Ser Trp Gly Gly
65 70 75 80
Ala Glu Pro His Glu Glu Gln Cys Leu Ser Ala Phe Thr Val His Phe
85 90 95
Ser Gly Gln Phe Thr Gly Thr Ala Gly Ala Cys Arg Tyr Gly Pro Phe
100 105 110
Gly Pro Pro Pro Pro Ser Gln Ala Ser Ser Gly Gln Ala Arg Met Phe
115 120 125
Pro Asn Ala Pro Tyr Leu Pro Ser Cys Leu Glu Ser Gln Pro Ala Ile
130 135 140
Arg Asr Gln Gly Tyr Ser Thr Val Thr Phe Asp Gly Thr Pro Ser Tyr
145 150 155 160
Gly His Thr Pro Ser His His Ala Ala Gln Phe Pro Asr His Ser Phe
165 170 175
Lys His Glu Asp Pro Met Gly Gln Gln Gly Ser Leu Gly Glu Gln Gln
180 185 190
Tyr Ser Val Pro Pro Pro Val Tyr Gly Cys His Thr Pro Thr Asp Ser
195 200 205
Cys Thr Gly Ser Gln Ala Leu Leu Leu Arg Thr Pro Tyr Ser Ser Asp
210 215 220
Asr Leu Tyr Gln Met Thr Ser Gln Leu Glu Cys Met Thr Trp Asn Gln
225 230 235 240
Met Asn Leu Gly Ala Thr Leu Lys Gly Val Ala Ala Gly Ser Ser Ser
245 250 255
Ser Val Lys Trp Thr Glu Gly Gln Ser Asn His Ser Thr Gly Tyr Glu
260 265 270
Ser Asp Asr His Thr Thr Pro Ile Leu Cys Gly Ala Gln Tyr Arg Ile
275 280 285
His Thr His Gly Val Phe Arg Gly Ile Gln Asp Val Arg Arg Val Pro
290 295 300
Gly Val Ala Pro Thr Leu Val Arg Ser Ala Ser Glu Thr Ser Glu Lys
305 310 315 320
Arg Pro Phe Met Cys Ala Tyr Pro Gly Cys Asn Lys Arg Tyr Phe Lys
325 330 335
Leu Ser His Leu Gln Met His Ser Arg Lys His Thr Gly Glu Lys Pro
340 345 350
Tyr Gln Cys Asp Phe Lys Asp Cys Glu Arg Arg Phe Ser Arg Ser Asp
355 360 365
Gln Leu Lys Arg His Gln Arg Arg His Thr Gly Val Lys Pro Phe Gln
370 375 380
Cys Lys Thr Cys Gln Arg Lys Phe Ser Arg Ser Asp His Leu Lys Thr
380 395 400
His Thr Arg Thr His Thr Gly Lys Thr Ser Glu Lys Pro Phe Ser Cys
405 4l0 415
Arg Trp Pro Ser Cys Gln Lys Lys Phe Ala Arg Ser Asp Glu Leu Val
420 425 430
Arg His His Asn MetHis Gln Arg Asn Met Thr Lys Leu Gln Leu Ala
435 440 445
Leu
<210>3
<211>8
<212>PRT
<213>人工序列
<220>
<223>合成肽
<400>3
Gly Ala Ser Ala Tyr Gly Ser Leu
1 5
<210>4
<211>8
<212>PRT
<213>人工序列
<220>
<223>合成肽
<400>4
Cys Asn Lys Arg Tyr Phe Lys Leu
1 5
<210>5
<211>9
<212>PRT
<213>人工序列
<220>
<223>合成肽
<400>5
Arg Met Phe Pro Asn Ala Pro Tyr Leu
1 5
<210>6
<211>9
<212>PRT
<213>人工序列
<220>
<223>合成肽
<400>6
Tyr Ser Ser Asp Asn Leu Tyr Gln Met
1 5
<210>7
<211>9
<212>PRT
<213>人工序列
<220>
<223>合成肽
<400>7
Cys Met Thr Trp Asn Gln Met Asn Leu
1 5
<210>8
<211>9
<212>PRT
<213>人工序列
<220>
<223>合成肽
<400>8
Ser Leu Gly Glu Gln G1n Tyr Ser Val
1 5
<210>9
<211>9
<212>PRT
<213>人工序列
<220>
<223>合成肽
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Cys Tyr Thr Trp Asn Gln Met Asn Leu
1 5
Claims (14)
1.含有以癌抑制基因WT1的产物、或其部分肽或修饰产物为活性成分的癌抗原和阳离子脂质体的癌疫苗;其中所述癌抗原为包含如序列3、4、5、6、7、8和9中任一序列所示的8-12个氨基酸的肽。
2.权利要求1记载的癌疫苗,其中癌是白血病、骨髓异形成综合征、恶性淋巴瘤、多发性骨髓瘤、胃癌、结肠癌、肺癌、乳腺癌、胚细胞癌、肝癌、皮肤癌、膀胱癌、***癌、子宫癌、子***、或卵巢癌。
3.权利要求1记载的癌疫苗,其中上述癌抗原肽是下列的任一种:
Kb45 Gly Ala Ser Ala Tyr Gly Ser Leu(序列3)
Kb330 Cys Asn Lys Arg Tyr Phe Lys Leu(序列4)
Db126 Arg Met Phe Pro Asn Ala Pro Tyr Leu(序列5)
Db221 Tyr Ser Ser Asp Asn Leu Tyr Gln Met(序列6)
Db235 Cys Met Thr Trp Asn Gln Met Asn Leu(序列7)
WH 187 Ser Leu Gly Glu Gln Gln Tyr Ser Val(序列8)。
4.权利要求3记载的癌疫苗,其中上述癌抗原肽是:
Db126 Arg Met Phe Pro Asn Ala Pro Tyr Leu(序列5)或
WH187 Ser Leu Gly Glu Gln Gln Tyr Ser Val(序列8)。
5.权利要求1记载的癌疫苗,其中上述癌抗原肽是含有如下氨基酸序列:Cys Tyr Thr Trp Asn Gln Met Asn Leu(序列9)的肽。
6.权利要求1~5中任一项记载的癌疫苗,其中上述阳离子脂质体是含有N-[1-(2,3-二油酰氧)丙基]-N,N,N-氯化三甲铵)、N-[1-(2,3-二油酰氧)丙基]-N,N,N-三甲铵硫酸甲酯、或双十八烷基酰胺甘氨酰精胺或它们与中性脂的混合物的脂质体。
7.权利要求6记载的癌疫苗,其中上述阳离子脂质体是脂质转染试剂Lipofectin。
8.以癌抑制基因WT1的产物或其部分肽或修饰物作为活性成分的癌抗原和阳离子脂质体的用途,用于制备癌疫苗;其中所述癌抗原为包含如序列3、4、5、6、7、8和9中任一序列所示的8-12个氨基酸的肽。
9.权利要求8记载的用途,其中癌是白血病、骨髓异形成综合征、恶性淋巴瘤、多发性骨髓瘤、胃癌、结肠癌、肺癌、乳腺癌、胚细胞癌、肝癌、皮肤癌、膀胱癌、***癌、子宫癌、子***、卵巢癌。
10.权利要求8记载的用途,其中上述癌抗原肽是下列的任一种:
Kb45 Gly Ala Ser Ala Tyr Gly Ser Leu(序列3)
Kb330 Cys Asn Lys Arg Tyr Phe Lys Leu(序列4)
Db126 Arg Met Phe Pro Asn Ala Pro Tyr Leu(序列5)
Db221 Tyr Ser Ser Asp Asn Leu Tyr Gln Met(序列6)
Db235 Cys Met Thr Trp Asn Gln Met Asn Leu(序列7)
WH187 Ser Leu Gly Glu Gln Gln Tyr Ser Val(序列8)。
11.权利要求10中记载的应用,其中上述癌抗原肽是
Db126 Arg Met Phe Pro Asn Ala Pro Tyr Leu(序列5)或
WH187 Ser Leu Gly Glu Gln Gln Tyr Ser Val(序列8)。
12.权利要求8记载的用途,其中上述癌抗原肽是含有如下氨基酸序列:Cys Tyr Thr Trp Asn Gln Met Asn Leu(序列9)的肽。
13.权利要求8~12任一项中记载的用途,其中上述阳离子脂质体是含有N-[1-(2,3-二油酰氧)丙基]-N,N,N-氯化三甲铵)、N-[1-(2,3-二油酰氧)丙基]-N,N,N-三甲铵硫酸甲酯、或双十八烷基酰胺甘氨酰精胺或他们与中性脂的混合物的脂质体。
14.权利要求13记载的用途,其中上述阳离子脂质体是脂质转染试剂Lipofectin。
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US (2) | US20060165708A1 (zh) |
EP (2) | EP2014300B1 (zh) |
JP (2) | JP4229832B2 (zh) |
CN (1) | CN1320923C (zh) |
AT (1) | ATE494905T1 (zh) |
CA (1) | CA2451846A1 (zh) |
DE (1) | DE60238958D1 (zh) |
HK (1) | HK1064589A1 (zh) |
WO (1) | WO2003002142A1 (zh) |
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EP2338509B1 (en) | 2003-06-27 | 2014-06-18 | International Institute of Cancer Immunology, Inc. | Method of selecting patients suitable for WT1 vaccine |
ATE470435T1 (de) * | 2003-11-03 | 2010-06-15 | Yissum Res Dev Co | Verfahren zur auswahl von kationischen oder anionischen liposomen zur behandlung einer schleimhautmembran und diese enthaltendes set |
KR20130062368A (ko) * | 2003-11-05 | 2013-06-12 | 인터내셔널 인스티튜트 오브 캔서 이무놀로지 인코퍼레이티드 | Wt1 유래의 hla-dr 결합성 항원 펩티드 |
US7303881B2 (en) * | 2004-04-30 | 2007-12-04 | Pds Biotechnology Corporation | Antigen delivery compositions and methods of use |
EP1657250A1 (en) * | 2004-11-11 | 2006-05-17 | Charité - Universitätsmedizin Berlin | HLA-A *01-binding T-cell epitope of WT1 |
CA2626238C (en) * | 2005-10-17 | 2015-10-06 | Sloan Kettering Institute For Cancer Research | Wt1 hla class ii-binding peptides and compositions and methods comprising same |
JP5002749B2 (ja) * | 2006-03-22 | 2012-08-15 | 富士フイルム株式会社 | 癌抑制剤 |
EP3834836A1 (en) | 2006-04-10 | 2021-06-16 | Memorial Sloan Kettering Cancer Center | Immunogenic wt-1 peptides and uses thereof |
JP2008031063A (ja) * | 2006-07-27 | 2008-02-14 | Fujifilm Corp | 癌抑制剤 |
NZ599161A (en) | 2007-02-27 | 2012-07-27 | Int Inst Cancer Immunology Inc | Method for activation of helper t cell and composition for use in the method |
WO2008105174A1 (ja) * | 2007-02-28 | 2008-09-04 | National University Corporation Hokkaido University | 細胞性免疫誘導用リポソーム |
EP3173480A3 (en) | 2007-03-05 | 2017-08-16 | International Institute of Cancer Immunology, Inc. | Cancer antigen-specific t-cell receptor gene, peptide encoded by the gene, and use of them |
US8877206B2 (en) * | 2007-03-22 | 2014-11-04 | Pds Biotechnology Corporation | Stimulation of an immune response by cationic lipids |
ITTO20070401A1 (it) * | 2007-06-07 | 2008-12-08 | Univ Degli Studi Torino | Vaccino anti-tumorale |
RU2530555C2 (ru) | 2008-04-17 | 2014-10-10 | ПиДиЭс БАЙОТЕКНОЛОДЖИ КОРПОРЭЙШН | Стимуляция иммунного ответа энантиомерами катионных липидов |
TW201623616A (zh) | 2010-10-05 | 2016-07-01 | 癌免疫研究所股份有限公司 | 用於活化細胞毒性t細胞的方法及組成物,及用於細胞毒性t細胞之活化誘導物 |
JP6082901B2 (ja) * | 2011-01-31 | 2017-02-22 | オリンパス株式会社 | ワクチン・アジュバント |
US9803246B2 (en) | 2011-06-28 | 2017-10-31 | International Institute Of Cancer Immunology, Inc. | Receptor gene for peptide cancer antigen-specific T cell |
EP2802347B1 (en) | 2012-01-13 | 2019-01-09 | Memorial Sloan Kettering Cancer Center | Immunogenic wt-1 peptides and methods of use thereof |
US8562681B2 (en) | 2012-01-31 | 2013-10-22 | Styker Spine | Laminoplasty implant, method and instrumentation |
TWI672149B (zh) | 2012-09-21 | 2019-09-21 | 美商Pds生技公司 | 改良之疫苗組成物及使用方法 |
NZ708990A (en) | 2012-12-17 | 2020-03-27 | Int Inst Cancer Immunology Inc | Method for activating helper t cell |
CN116789792A (zh) | 2013-01-15 | 2023-09-22 | 纪念斯隆凯特林癌症中心 | 免疫原性wt-1肽和其使用方法 |
US10815273B2 (en) | 2013-01-15 | 2020-10-27 | Memorial Sloan Kettering Cancer Center | Immunogenic WT-1 peptides and methods of use thereof |
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CN1520311A (zh) | 2004-08-11 |
EP1410804A1 (en) | 2004-04-21 |
EP2014300A1 (en) | 2009-01-14 |
US20060165708A1 (en) | 2006-07-27 |
WO2003002142A1 (en) | 2003-01-09 |
JP4229832B2 (ja) | 2009-02-25 |
JP2009079054A (ja) | 2009-04-16 |
HK1064589A1 (en) | 2005-02-04 |
ATE494905T1 (de) | 2011-01-15 |
EP1410804A4 (en) | 2007-10-24 |
EP2014300B1 (en) | 2011-01-12 |
US20090098197A1 (en) | 2009-04-16 |
WO2003002142A8 (en) | 2003-12-11 |
DE60238958D1 (de) | 2011-02-24 |
JPWO2003002142A1 (ja) | 2004-10-14 |
CA2451846A1 (en) | 2003-01-09 |
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