相关申请的交叉引用本专利申请要求于2017年9月29日提交的美国临时专利申请号62/565,383的权益，其通过引用整体并入本文。关于联邦政府资助的研究和开发的声明本发明是由美国国立卫生研究院国家癌症研究所(nationalinstitutesofhealth,nationalcancerinstitute)在政府支持下完成的，项目号为bc010985。政府拥有本发明的某些权利。通过引用并入以电子方式提交的材料通过引用整体并入本文的是与此同时提交并且标识如下的计算机可读核苷酸/氨基酸序列表:日期为2018年9月7日的一个名为“739980_st25.txt”的687,555字节ascii(文本)文件。发明背景某些癌症的治疗选择可能非常有限，尤其是当癌症转移且无法切除时。尽管例如手术、化疗和放射治疗等治疗取得了进展，但许多癌症(例如胰腺、结直肠、肺、子宫内膜、卵巢和前列腺癌)的预后可能较差。因此，对癌症额外治疗的需求尚未得到满足。发明简述本发明的一个实施方案提供对突变的人p53具有抗原特异性的分离或纯化t细胞受体(tcr)。本发明的其它实施方案提供涉及本发明tcr的相关多肽和蛋白质以及相关核酸、重组表达载体、宿主细胞、细胞群以及药物组合物。本发明的另外实施方案提供了检测哺乳动物中癌症存在的方法以及治疗或预防哺乳动物中癌症的方法。若干附图视图的简述图1是显示与用所指示的浓度(μg/ml)渐低的wtp53-g245肽(wt)(无阴影条)或突变型p53-g245s肽(mut)(阴影条)脉冲的自体树突状细胞(dc)共培养后，在与用来自患者4127的一个指定tcr转导的自体pbl共培养后测量的ifn-γ(pg/ml)浓度的图。图2是显示在将用来自患者4127的一个所示tcr转导的自体pbl与用所指示的浓度(μg/ml)渐低的wtp53-g245肽(wt)(无阴影条)或突变型p53-g245s肽(mut)(阴影条)脉冲的自体树突状细胞(dc)共培养后，检测到的4-1bb阳性细胞的百分比(％cd4+mtcrβ+)的图。图3是显示在将用来自患者4127的一个所示tcr转导的自体pbl与用表a所示的一种肽脉冲的自体dc共培养后，检测到的4-1bb阳性细胞的百分比(％cd4+mtcrβ+)的图。图4是显示在将来自患者4127的til或用一个所示tcr转导的t细胞与cos7细胞共培养后测量的ifn-γ(pg/ml)浓度的图，所述cos7细胞用一种所示hla等位基因转导并且用dmso或一种所示肽脉冲。图5是显示在将来自患者4127的til或用一个所示tcr转导的t细胞与cos7细胞共培养后检测到的4-1bb阳性细胞(cd4+mtcrβ+的％)的百分比的图，所述cos7细胞用一种所示hla等位基因转导并且用dmso或一种所示肽脉冲。图6是显示在共培养来自患者4127的til与同种异体的(drb3*01:01:01或drb3*02:02:01)apc之后测量的每2x104个效应细胞的斑点的数量的图，所述同种异体的(drb3*01:01:01或drb3*02:02:01)apc(1)用由无关(irr；左侧斜线空条)、wtp53(p53-wt；左侧斜线灰条)或突变的p53(p53-mut；灰条)序列组成的tmg电穿孔或(2)用肽媒介物(dmso；右侧斜线空条)或由wtp53-g245序列(lp-p53-wt-g245；右侧斜线灰条)或突变的p53-g245s(lp-p53-mut-g245s；黑条)序列组成的经纯化的(>95％，通过hplc)25个氨基酸的肽脉冲。图7是显示在共培养表达4127-tcr1的细胞与同种异体的(drb3*01:01:01或drb3*02:02:01)apc之后测量的每2x104个效应细胞的斑点的数量的图，所述同种异体的(drb3*01:01:01或drb3*02:02:01)apc(1)用由无关(irr；左侧斜线空条)、wtp53(p53-wt；左侧斜线灰条)或经突变的p53(p53-mut；灰条)序列组成的tmg电穿孔或(2)用肽媒介物(dmso；右侧斜线空条)或由wtp53-g245序列(lp-p53-wt-g245；右侧斜线灰条)或经突变的p53-g245s(lp-p53-mut-g245s；黑条)序列组成的经纯化的(>95％，通过hplc)25个氨基酸的肽脉冲。图8a是显示将来自用于治疗患者4196的输注袋的til与转染了tmg1的质粒和指定hla等位基因的cos7细胞共培养后测量的ifn-γ分泌(斑点/2×104个细胞)的图。图8b是显示将来自用于治疗患者4196的输注袋的til与用候选最小表位(hmtevvrhc(seqidno:530)或(sqhmtevrh(seqidno:531))脉冲的自体dc共培养后测量的ifn-γ分泌(斑点/2×104个细胞)的图。单独培养的t细胞作为对照。图9a-9d是显示用指示浓度(μg/ml)的最小wt(圆圈)或突变的(mut)(方块)表位脉冲的dc与使用以下tcr之一转导的细胞共同培养后检测到的mtcrβ+cd8+41bb+细胞的百分比的图:av12/bv6-1(a)，av2/bv6-1(b)、av6/bv11(c)或av38/bv10-3(d)。将脉冲dcs与未转导(ut)细胞(三角形)共培养作为对照。显示的结果代表了两次实验。图10a-10b是显示用指示浓度(μg/ml)的最小突变的表位脉冲的t2细胞与使用以下tcr之一转导的细胞共同培养后测得的ifn-γ分泌(pg/ml)(a)和4-1bb阳性细胞百分比(％，mtcrβ+)(b)的图:av12/bv6-1(实心三角形)，av6/bv11(圆形)或av38/bv10-3(方形)。将脉冲的t2细胞与未转导(ut)细胞(空心三角形)共培养作为对照。图11是显示将tcr转导细胞与指定靶细胞系共培养后，检测到的每2x104个效应细胞的ifn-γ阳性斑点数的图。用av12/bv6-1(黑色条)、av6/bv11(条纹条)或av38/bv10-3(灰色条)转导效应细胞。hla-a*02:01转导的靶细胞由ls123-a2、ccrf-cem-a2和au-565-a2指示。肽脉冲的靶细胞表示为(+)。未用肽脉冲的靶细胞用(-)表示。使用与靶细胞系共培养的未转导效应细胞(无阴影条)作为对照。图12是显示在患者4238til片段f1-f24(n＝24)与用由不相关的(irr；空心条)或突变型p53(p53-mut；黑条)序列组成的tmg电穿孔的自体apc共培养后，检测到的每2x104个效应细胞的ifn-γ阳性斑点数的图。图13是显示在患者4238til片段f1-f24(n＝24)与用肽媒介物(dmso；空心条)或突变p53-r248q(lp-p53-r248q-mut；黑条)序列组成的纯化(hplc法>95％)的25个氨基酸的肽脉冲的自体apc共培养后，检测到的每2x104个效应细胞的ifn-γ阳性斑点数的图。图14是显示患者4238til片段f1-f24(n＝24)与用不相关的(irr；空心条)或突变型p53(p53-mut；黑条)序列组成的tmg电穿孔的自体apc共培养后，检测到的cd8+4-1bb+细胞的百分比的图。图15是显示4238til片段f1-f24(n＝24)与用肽媒介物(dmso；空心条)或突变p53-r248q(lp-p53-r248q-mut；黑条)序列组成的纯化(hplc法>95％)的25个氨基酸的肽脉冲的自体apc共培养后，检测到的cd8+4-1bb+细胞的百分比的图。图16是显示在共培养来自患者4253的til片段(f1-f24，n＝24)与自体apc之后测量的每2x104个效应细胞的ifn-γ-阳性点的数量的图，所述自体apc(1)用肽媒介物(dmso；空条)脉冲，(2)用由突变的p53r248w(lp-p53r248w-mut；黑条)序列组成的经纯化的(>95％，通过hplc)25-个氨基酸的肽脉冲，(3)用无关tmg电穿孔(tmg-irr；灰斜线条)、或(4)含突变p53-r248w序列的p53-mut-tmg电穿孔。图17是显示在共培养来自患者4273的til片段(f1-f24，n＝24)与自体apc之后测量的每2x104个效应细胞的ifn-γ-阳性点的数量的图，所述自体apc用由无关(irr；空条)、wtp53(p53-wt；灰条)或经突变的p53(p53-mut；黑条)序列组成的tmg电穿孔。图18是显示在共培养来自患者4273的til片段(f1-f24，n＝24)与自体apc之后检测的cd4+4-1bb+细胞的百分比的图，所述自体apc用由无关(irr；空条)、wtp53(p53-wt；灰条)或经突变的p53(p53-mut；黑条)序列组成的tmg电穿孔。图19是显示在共培养来自患者4273的til片段(f1-f24，n＝24)与自体apc之后测量的每2x104个效应细胞的ifn-γ-阳性斑点的数量的图，所述自体apc用肽媒介物(dmso；空条)或由wtp53-r248序列(lp-p53-r248-wt；灰条)或经突变的p53-r248w(lp-p53-r248w-mut；黑条)序列组成的经纯化的(>95％，通过hplc)25-个氨基酸的肽脉冲。图20显示在共培养来自患者4273的til片段(f1-f24，n＝24)与自体apc之后检测的cd4+4-1bb+细胞的百分比的图，所述自体apc用肽媒介物(dmso；空条)或由wtp53-r248序列(lp-p53-r248-wt；灰条)或经突变的p53-r248w(lp-p53-r248w-mut；黑条)序列组成的经纯化的(>95％，通过hplc)25-个氨基酸的肽脉冲。图21是显示在共培养来自患者4149的自体pbl(经4149-tcra2b1或4149-tcra2b2转座)与自体apc之后测量的每2x104个效应细胞的ifn-γ-阳性斑点的数量的图，所述自体apc(1)用由无关的(irr，右侧斜线黑条)、wtp53(p53-wt-tmg,左侧斜线黑条)或经突变的p53(p53-mut-tmg，灰条)序列组成的tmg电穿孔，或(2)用肽媒介物(dmso，空心条)或由wtp53-y220序列(lp-p53-y220-wt，水平阴影黑条)或经突变的p53-y220c(lp-p53-y220c-mut，黑条)序列组成的经纯化的(>95％，通过hplc)25-个氨基酸的肽脉冲。佛波醇12-肉豆蔻酸13-乙酸酯(pma)和离子霉素(iono)为阳性对照(灰色条)。图22是显示在共培养来自患者4149的自体pbl(其用tcr(4149-tcra2b1或4149-tcra2b2)转座)与自体apc共培养之后检测的cd4+4-1bb+细胞的百分比的图，所述自体apc(1)用由无关的(irr，右侧斜线黑条)、wtp53(p53-wt-tmg,左侧斜线黑条)或经突变的p53(p53-mut-tmg，灰条)序列组成的tmg电穿孔，或(2)用肽媒介物(dmso，空心条)或由wtp53-y220序列(lp-p53-y220-wt，水平阴影黑条)或经突变的p53-y220c(lp-p53-y220c-mut，黑条)序列组成的经纯化的(>95％，通过hplc)25-个氨基酸的肽脉冲。pma和iono为阳性对照(灰色条)。图23是显示来自患者4149的til与用表f的其中一个肽脉冲的自体dc共同培养后检测到的4-1bb+细胞的百分比(cd4+t细胞百分比)的图。图24是显示4149-tcra2b2转座的t细胞与用单个hla等位基因+/-tmg共转染的cos7细胞共培养后的ifn-γ分泌(pg/ml)的图。未转染tmg的细胞用p53y220c15-mer肽脉冲。脉冲的靶细胞用阴影条表示。用无阴影条指示tmg转染的靶细胞。图25是显示患者4213的til片段(f2和f24)与自体apc共培养之后检测的cd8+4-1bb+细胞的百分比的图，所述自体apc用肽媒介物(dmso，空心条)或由突变的p53-r248q(lp-p53-r248q-mut；黑条)序列组成的经纯化的(>95％，通过hplc)25-个氨基酸的肽脉冲。图26是患者4213的til片段(f2和f24)与自体apc共培养之后测量的每2x104个效应细胞的ifn-γ-阳性斑点的数量的图，所述自体apc用肽媒介物(dmso，空心条)或由突变的p53-r248q(lp-p53-r248q-mut；黑条)序列组成的经纯化的(>95％，通过hplc)25-个氨基酸的肽脉冲。ifn-γ的分泌由空心条表示。4-1bb的表达由实心条显示。图27是患者4268的til片段(f1-f24，n＝24)与自体apc共培养之后测量的每2x104个效应细胞的ifn-γ-阳性斑点的数量的图，所述自体apc用由无关的(irr，空心条)、wtp53(p53-wt,灰条)或经突变的p53(p53-mut，黑条)序列组成的tmg电穿孔。图28是显示患者4268的til片段(f1-f24，n＝24)与自体apc共培养之后测量的每2x104个效应细胞的ifn-γ-阳性斑点的数量的图，所述自体apc用肽媒介物(dmso，空心条)或由wtp53-r248序列(lp-p53-r248-wt；灰色条)或突变的p53-r248q(lp-p53-r248q-mut；黑条)序列组成的经纯化的(>95％，通过hplc)25-个氨基酸的肽脉冲。图29是显示患者4268的til片段(f1-f24，n＝24)与自体apc共培养之后检测的cd4+4-1bb+细胞的百分比的图，所述自体apc用肽媒介物(dmso，空心条)或由wtp53-r248序列(lp-p53-r248-wt；灰色条)或突变的p53-r248q(lp-p53-r248q-mut；黑条)序列组成的经纯化的(>95％，通过hplc)25-个氨基酸的肽脉冲。图30是显示患者4268的til片段(f1-f24，n＝24)与自体apc共培养之后检测的cd8+4-1bb+细胞的百分比的图，所述自体apc用肽媒介物(dmso，空心条)或由wtp53-r248序列(lp-p53-r248-wt；灰色条)或突变的p53-r248q(lp-p53-r248q-mut；黑条)序列组成的经纯化的(>95％，通过hplc)25-个氨基酸的肽脉冲。图31是显示在来自患者4266的til片段(f1-f24，n＝24)与自体apc共培养之后测量的每2x104个效应细胞的ifn-γ-阳性斑点的数量的图，所述自体apc用由无关的(irr，空心条)、wtp53(p53-wt，灰条)或经突变的p53(p53-mut，黑条)序列组成的tmg电穿孔。图32是显示在来自患者4266的til片段(f1-f24，n＝24)与自体apc共培养之后检测的cd8+4-1bb+细胞的百分比的图，所述自体apc用由无关的(irr，空心条)、wtp53(p53-wt，灰条)或经突变的p53(p53-mut，黑条)序列组成的tmg电穿孔。图33是显示在来自患者4266的til片段(f1-f24，n＝24)与自体apc共培养之后测量的每2x104个效应细胞的ifn-γ-阳性斑点的数量的图，所述自体apc用肽媒介物(dmso，空心条)或由wtp53-r248序列(lp-p53-r248-wt；灰色条)或突变的p53-r248w(lp-p53-r248w-mut；黑条)序列组成的经纯化的(>95％，通过hplc)25-个氨基酸的肽脉冲。图34是显示在来自患者4266的til片段(f1-f24，n＝24)与自体apc共培养之后检测的cd8+4-1bb+细胞的百分比的图，所述自体apc用肽媒介物(dmso，空心条)或由wtp53-r248序列(lp-p53-r248-wt；灰色条)或突变的p53-r248w(lp-p53-r248w-mut；黑条)序列组成的经纯化的(>95％，通过hplc)25-个氨基酸的肽脉冲。图35显示了九种p53剪接变体的氨基酸序列的比对。sp|p04637|p53_human(seqidno:1)；sp|p04637-2|p53_human(seqidno:535)；sp|p04637-3|p53_human(seqidno:536)；sp|p04637-4|p53_human(seqidno:537)；sp|p04637-5|p53_human(seqidno:538)；sp|p04637-6|p53_human(seqidno:539)；sp|p04637-7|p53_human(seqidno:540)；sp|p04637-8|p53_human(seqidno:541)；和sp|p04637-9|p53_human(seqidno:542)。图36是显示在与4127-o37-tcr转导的t细胞与自体apc共培养后检测到的4-1bb阳性细胞百分比(mtcrβ+的％)的图，所述apc用对应于wt(空心圆)或突变(实心方形)p53–g245s序列的浓度渐低的25个氨基酸的肽脉冲。图37a-37c是显示在将4127-o37-tcr(a)、4127-tp31-g245s-tcr1(b)或4127-o102-tcr(c)转导的t细胞与cos7细胞共同培养后测量的4-1bb阳性细胞百分比(mtcrβ+的％)(左y轴；斜线条)和ifn-γ(pg/ml)(右y轴；黑条)的图，所述cos7细胞用一种所示的hla等位基因进行共转染并用25-氨基酸的p53-g245s肽脉冲。图38是显示在将患者4141的til(片段培养物12)与自体apc共培养后测量的4-1bb阳性细胞百分比(％cd8+)的图形(右y轴；黑条)和ifn-γ(斑点数/2x104个细胞)(左y轴；斜线条)的图，所述apc用编码无关突变(tmg-irr)、wtp53序列(tp53-wt-tmg)或突变p53序列(包括r175h)(tp53-mut-tmg)的tmg转染。单独的培养基以及pma和离子霉素分别为阴性和阳性对照。图39为显示将患者4141的til(片段培养物12)与cos7细胞共培养之后测量的每2x104个效应细胞的ifn-γ-阳性斑点的数量的图，所述cos7细胞用所示的hla等位基因并无额外基因(仅hla；空心条)共转染、或者用wttp53tmg(灰色阴影条)或含有p53-r175h序列的突变(黑色条)tp53tmg共转染。图40为显示将表达模拟物(无tcr)或4141-tcr1a2的t细胞与t2肿瘤细胞(表达hla-a*02:01)共培养后测定的ifn-γ浓度(pg/ml)的图。t2细胞用肽媒介物(dmso；灰色条)或纯化(hplc法>95％)的由wtp53-r175肽(阴影灰色条)或突变p53-r175h肽(黑色条)组成的肽脉冲。仅培养基(空心条)以及pma和离子霉素(格状条)分别为阴性和阳性对照。数据为平均值±sem(n＝3)。图41是显示将表达4141-tcr1a2的t细胞与saos2细胞(p53-null和hla-a*02:01+)共培养后阳性表达一种所示标记物的细胞百分比的图，这些细胞要么未经处理(无阴影条)，要么被使得过度表达全长p53r-175h蛋白(阴影条)。数据为平均值±sem(n＝3)。对两种细胞系之间的每种细胞因子进行student双尾t检验，以进行统计分析(***p<0.001)。图42是显示在将til(来自患者4259的片段培养物6)与自体apc共培养后检测到的cd4+4-1bb阳性细胞的百分比的图，所述apc或者(1)用由无关的(tmg-irr)、wtp53(tmg-p53-wt)或经突变的p53(tmg-p53-mut)序列组成的tmg电穿孔，或者(2)用肽媒介物(dmso)或由wtp53-y220序列(lp-p53-y220-wt)或突变的p53-y220c(lp-p53-y220c-mut)序列组成的纯化的(hplc法>95％)25-氨基酸肽脉冲。图43是显示将来自患者4259的til片段培养物(no.6)与自体apc共培养后检测到的cd4+4-1bb阳性细胞的百分比的图，所述apc用对应于wtp53序列(空心圆)或突变的p53-y220c(实心方形)的浓度逐渐降低的25-氨基酸肽在37oc下脉冲2小时。图44是显示患者4259的til与用dmso、wtp53-y220肽或突变p53-y220c肽脉冲的自体apc共培养后检测到的cd4+4-1bb阳性细胞的百分比的图。图45是显示将来自患者4259的til片段培养物(no.6)与cos7细胞共培养后检测到的4-1bb阳性细胞百分比(cd4+％)的图，所述cos7细胞用来自患者4259的所示hla等位基因共转染，并用dmso(空心条)或p53-y220c肽(实心条)脉冲。图46是tc#4259靶细胞(内源性表达p53y220c和hla-drb1*04:01:01)与表达模拟物(无tcr，空心条)或p53-y220c特异性tcr(4259-f6-tcr；黑条)的效应t细胞(105)共培养后检测到的4-1bb+细胞百分比的图。将tc#4259细胞不与任何物质孵育、或将tc#4259细胞与w6/32泛-hlai类特异性阻断抗体、iva12泛-hlaii类特异性阻断抗体或经突变的p53-y220c肽孵育。单独的培养基(无tc)及pma和离子霉素分别是阴性对照和阳性对照。数据是平均值±sem(n＝3)。在指定的组之间进行学生双尾t检验用于统计分析(**p<0.01和***p<0.001)。图47是显示将来自患者4285的til片段(f1-f22和f24，n＝23)与自体apc共培养之后检测的每2x104个效应细胞的ifn-γ-阳性斑点的数量的图，所述自体apc用由无关的(irr，空心条)、wtp53(p53-wt，灰条)或经突变的p53(p53-mut，黑条)序列组成的tmg电穿孔。图48是显示将来自患者4285的til片段(f1-f22和f24，n＝23)与自体apc共培养之后检测的每2x104个效应细胞的ifn-γ-阳性斑点的数量的图，所述apc用肽媒介物(dmso；空心条)或由wtp53-r175序列(lp-p53-r175-wt；灰色条)或突变型p53-r175h(lp-p53-r175h-mut；黑条)组成的经纯化(hplc法>95％)的25个氨基酸的肽脉冲。图49是显示将来自患者4285的til片段(f1-f22和f24，n＝23)与自体apc共培养之后检测的cd4+4-1bb阳性细胞的百分比的图，所述自体apc用由无关的(irr，空心条)、wtp53(p53-wt，灰条)或经突变的p53(p53-mut，黑条)序列组成的tmg电穿孔。。图50是显示将来自患者4285的til片段(f1-f22和f24，n＝23)与自体apc共培养之后检测的cd4+4-1bb阳性细胞的百分比的图，所述apc用肽媒介物(dmso；空心条)或由wtp53-r175序列(lp-p53-r175-wt；灰色条)或突变型p53-r175h(lp-p53-r175h-mut；黑条)组成的经纯化(hplc法>95％)的25个氨基酸的肽脉冲。图51是显示在共培养til片段培养物4285-f6、4285-f9和4285-f10与cos7细胞之后检测的4-1bb+细胞的百分比(cd4+百分比)的图，所述cos7细胞用指定hla等位基因转染并用dmso(肽媒介物；灰和黑斜线条)或突变的p53-r175h肽(灰、格子和黑条)脉冲。图52是显示在共培养用4285-tcr1转座的t细胞与自体apc之后检测的4-1bb+细胞的百分比(cd4+百分比)的图，所述自体apc用降低浓度的25-个或15-个氨基酸的肽(对应于wt(空心环和方块)或经突变的(实心环和方块)p53-r175h序列)脉冲。图53是显示在共培养来自患者4266的til与cos7细胞之后检测的4-1bb+细胞的百分比(cd8+百分比)的图，所述cos7细胞用来自患者4266的单独hla等位基因共转染以及用无肽(空条)、dmso(肽媒介物；灰条)、wtp53-r248肽(灰斜线条)或经突变的p53-r248w肽(黑条)脉冲。数据是平均值±sem(n＝3)。图54是显示在共培养表达模拟物(无tcr)、4266-tcr1、4266-tcr2、4266-tcr3或4266-tcr4的t细胞与自体apc之后检测的4-1bb+细胞的百分比(cd8+百分比)的图，所述自体apc用肽媒介物(dmso；灰条)或由wtp53-r248序列(斜线灰条)或经突变的p53-r248w(黑条)序列组成的经纯化的(>95％，通过hplc)25个氨基酸的肽脉冲。单独的培养基(空条)及pma和离子霉素(格条)分别是阴性对照和阳性对照。图55是显示在共培养表达模拟物(无tcr)或p53-r248w-特异性tcr(4266-tcr2、4266-tcr3或4266-tcr4)的t细胞与从由患者4266切除的异种移植的肿瘤碎片建立然后通过免疫受损小鼠(tc#4266)连续传代的肿瘤细胞(tc)系之后检测的4-1bb+细胞的百分比(cd8+百分比)的图。将tc#4266细胞不与任何物质孵育(黑条)、或将tc#4266细胞与w6/32泛-hlai类特异性阻断抗体(右侧灰斜线条)、iva12泛-hlaii类特异性阻断抗体(灰条)或突变的p53-r248w肽(左侧灰斜线条)孵育。单独的培养基(无tc；空条)及pma和离子霉素(灰格条)分别是阴性对照和阳性对照。图56是显示在来自患者4273的til与自体apc共培养后检测到的cd4+4-1bb+细胞的百分比的图，所述apc用编码不相关突变(灰色条)、野生型p53序列(灰色斜线条)或包括p53-r248w的突变p53序列(黑色条)的tmg转染。单独的培养基(空心条)以及pma和离子霉素(格状条)分别为阴性和阳性对照。图57是显示患者4273的til与自体apc共培养后检测到的cd4+4-1bb+细胞百分比的图，所述apc用对应于野生型(空心环)的25个氨基酸的肽或来自p53-r248w新表位的突变(实心方形)的25个氨基酸的肽脉冲。dmso为肽媒介物。图58是显示患者4273的til与自体apc共培养后检测到的cd4+4-1bb+细胞的百分比的图，所述apc用重叠14个氨基酸的来自p53-r248w新表位的15个氨基酸肽脉冲(以粗体表示氨基酸取代)。dmso为肽媒介物，单独的培养基(仅t细胞)，以及pma和离子霉素为对照。25个氨基酸肽(wtp53-r248和突变的p53-r248w)是15个氨基酸肽的额外对照。肽为:图59是显示来自患者4273的til与cos7细胞共培养之后分泌的ifn-γ(pg/ml)的浓度的图，所述cos7细胞用来自患者4273的单独hla等位基因以及分别在有或无p53-r248w新抗原的情况下的wt(空条)或突变的(黑条)tp53tmg共转染。图60是显示在表达模拟物(无tcr)或4273-tcr1a2的t细胞与自体apc的共培养之后测量的每2x104个细胞的ifn-γ-点的数量的图，所述自体apc用肽媒介物(dmso；灰条)或由wtp53-r248序列(斜线灰条)或经突变的p53-r248w(黑条)序列组成的经纯化的(>95％，通过hplc)25个氨基酸的肽脉冲。单独的培养基(空条)及pma和离子霉素(格条)分别是阴性对照和阳性对照。发明详述肿瘤蛋白p53(也称为“tp53”或“p53”)通过例如调节细胞分裂而作为肿瘤遏制物。p53蛋白位于细胞核中，在那里其直接结合至dna。当dna受损时，p53蛋白参与确定dna是否会被修复或受损的细胞是否会发生凋亡。如果dna可以被修复，则p53激活其他基因以恢复损伤。如果dna无法被修复，则p53蛋白阻止细胞分裂并向其发出进行凋亡的信号。通过阻止具有突变或受损dna的细胞分裂，p53有助于预防肿瘤发展。野生型(wt)全长p53包含seqidno:1的氨基酸序列。p53蛋白中的突变可降低或消除p53蛋白的肿瘤遏制功能。可替代地或另外地，p53突变可以是通过以显性负性方式干扰野生型p53的功能获得性突变。经突变的p53蛋白可以在多种人类癌症中的任一种中表达，诸如例如，胆管癌、黑素瘤、结肠癌、直肠癌、卵巢癌、子宫内膜癌、非小细胞肺癌(nsclc)、成胶质细胞瘤、子宫颈癌、头颈癌、乳房癌、胰腺癌或膀胱癌。本发明的一个实施方案提供一种对突变的人p53具有抗原性特异性的分离或纯化的t细胞受体(tcr)(下文称为“突变的p53”)。下文中，除非另有规定，否则提及“tcr”亦指tcr的功能部分及功能变体。p53的突变在本文中通过参考全长氨基酸酸序列wtp53(seqidno:1)来定义。因此，p53的突变在本文中通过参考在特定位置存在的氨基酸残基，然后是位置编号，然后是在所讨论的特定突变中取代此残基的氨基酸来描述。例如，当位置如由seqidno:1所定义时，术语“r175”是指存在于seqidno:1的位置175处的精氨酸，“r175h”表示存在于seqidno:1的位置175处的精氨酸被组氨酸替代，而“g245s”表示存在于seqidno:1的位置245处的甘氨酸已经被丝氨酸替代。p53具有九种已知的剪接变体。本文所述的p53突变在所有九种p53剪接变体中是保守的。图35中显示了9种p53剪接变体的比对。因此，通过本发明的方法分离的t细胞可对由九种p53剪接变体中任一种编码的本文所述的任何经突变的p53氨基酸序列具有抗原特异性。当位置如seqidno:1所定义时，则p53的特定剪接变体的氨基酸序列的实际位置是相对于seqidno:1的相应位置来定义的，并且如由seqidno:1定义的位置可与特定剪接变体中的实际位置不同。因此，例如，突变是指对应于seqidno:1的393个氨基酸的序列的指定位置的p53的特定剪接变体的氨基酸序列中的氨基酸残基的替代，要理解剪接变体中的实际位置可有所不同。在本发明的一个实施方案中，tcr对在seqidno:1的位置175、220、245、248、249、273或282处具有突变的人p53具有抗原特异性。tcr可对在seqidno:1的位置175、220、245或248具有突变的人p53具有抗原特异性。p53突变可能是任何错义突变。因此，seqidno:1的位置175、220、245、248、249、273或282处的突变可以是seqidno:1的位置175、220、245、248、249、273或282处存在的天然(wt)氨基酸残基的取代，其中在所讨论的特定位置处存在除天然(wt)氨基酸残基以外的任何氨基酸残基。在本发明的一个实施方案中，tcr对具有以下一种人p53突变的人p53具有抗原性特异性:r175h、y220c、g245d、g245s、r248l、r248q、r248w、r249s、r273h、r273c、r273l或r282w。seqidnoseqidnotcr可对具有以下一种人p53突变的人p53具有抗原特异性:r175h、y220c、g245s、r248q或r248w。例如，本发明tcr可对选自seqidno:2-13的突变型p53氨基酸序列具有抗原特异性。在本发明的一个实施方案中，本发明tcr可以能够以hla(人类白细胞抗原)分子依赖性方式识别突变型p53。本文使用的“hla-分子依赖性方式”意指tcr在hla分子背景下结合突变型p53时产生免疫应答，hla分子由从中分离出tcr的患者表达。本发明tcr可以能够识别由适用的hla分子呈递的突变型p53，且除突变型p53外还可与hla分子结合。本发明的tcr提供许多优点，包括当由用于过继细胞转移的细胞表达时。突变型p53由肿瘤细胞表达，而不是由正常的非癌细胞表达。在不约束于特定理论或机制的情况下，认为本发明tcr有利地靶向破坏癌细胞，同时尽可能减少或消除正常非癌细胞的破坏，从而例如通过最小化或消除毒性而减少毒性。此外，有利地，本发明tcr可成功地治疗或预防对其他类型的治疗(例如，化疗、手术或辐射)无响应的突变p53阳性癌症。此外，本发明tcr可提供对突变p53的高亲和识别，其可提供识别未经处理的肿瘤细胞的能力(例如，未经干扰素(ifn)-γ处理的肿瘤细胞，未用编码一种或两种突变p53和适用的hla分子的载体转染的肿瘤细胞，未用带有p53突变的p53肽脉冲的肿瘤细胞，或其组合)。大约一半的肿瘤携带p53突变，其中约一半为错义突变，约30％的错义突变发生在以下“热点”残基:r175h、y220c、g245d、g245s、r248l、r248q、r248w、r249s、r273c、r273l、r273h和r282w。此外，相同的p53“热点”突变(例如，r175h、y220c、g245d、g245s、r248l、r248q、r248w、r249s、r273h、r273c、r273l或r282w)常发生于无关人群的肿瘤中(累积约30％的p53错义突变)。因此，本发明tcr可以增加可能适合免疫疗法治疗的患者人数。如本文所使用的短语“抗原特异性”意指tcr可特异性结合并免疫识别具有高亲和力的突变p53。例如，如果表达tcr的约1x104至约1x105t细胞与以下抗原阴性、适用hla分子阳性的靶细胞共培养时分泌至少约200pg/ml或更高(例如，200pg/ml或更高、300pg/ml或更高、400pg/ml或更高、500pg/ml或更高、600pg/ml或更高、700pg/ml或更高、1000pg/ml或更高、5,000pg/ml或更高、7,000pg/ml或更高、10,000pg/ml或更高、20,000pg/ml或更高，或由前述值中的任两个所限定的范围)的ifn-γ，则所述tcr被视为对突变的p53具有“抗原特异性”:(a)用低浓度的突变p53肽(例如，约0.05ng/ml至约5ng/ml、0.05ng/ml、0.1ng/ml、0.5ng/ml、1ng/ml、5ng/ml，或由前述值中的任两个定义的范围)脉冲的抗原阴性的、适用hla分子阳性的靶细胞，或(b)已引入编码突变p53的核苷酸序列以使靶细胞表达突变p53的抗原阴性、适用的hla分子阳性的靶细胞。表达本发明tcr的细胞也可在与用较高浓度的突变p53肽脉冲的抗原阴性、适用hla分子阳性靶细胞共培养时分泌ifn-γ。可选地或者另外地，如果与阴性对照表达ifn-γ的量相比，表达tcr的t细胞与(a)用低浓度突变型p53肽脉冲的抗原阴性、适用hla分子阳性的靶细胞或(b)已引入编码突变型p53的核苷酸序列以使靶细胞表达突变p53的的抗原阴性、适用hla分子阳性的靶细胞共同培养时分泌至少两倍的ifn-γ，则tcr可被视为对突变型p53具有“抗原特异性”。阴性对照可以是，例如(i)与以下靶细胞共培养的表达tcr的t细胞:(a)用相同浓度的不相关的肽(例如，与突变型p53肽具有不同序列的一些其他肽)脉冲的抗原阴性、适用hla分子阳性的靶细胞或(b)已引入编码不相关肽的核苷酸序列以使靶细胞表达不相关肽的抗原阴性、适用hla分子阳性的靶细胞，或(ii)与以下靶细胞共培养的未转导的t细胞(例如来自pbmc，其不表达tcr):(a)与相同浓度的突变p53肽脉冲的抗原阴性、适用hla分子阳性靶细胞或(b)已引入编码突变p53的核苷酸序列以使靶细胞表达突变的p53的抗原阴性、适用hla分子阳性的靶细胞。ifn-γ分泌可藉由本领域已知的方法(例如，酶联免疫吸附测定(elisa))来测量。可选地或者另外地，如果与分泌ifn-γ的阴性对照t细胞的数量相比，至少两倍数量的表达tcr的t细胞与以下靶细胞共培养时分泌ifn-γ，则所述tcr可被视为对突变型p53具有“抗原特异性”:(a)用低浓度突变型p53肽脉冲的抗原阴性、适用hla分子阳性靶细胞或(b)已引入编码突变型p53的核苷酸序列以使靶细胞表达突变的p53的抗原阴性、适用hla分子阳性的靶细胞。肽和阴性对照的浓度可如本文关于本发明的其它方面所述。可藉由本领域的已知方法(例如，酶联免疫斑点(elisot)分析)来测量分泌ifn-γ的细胞的数量。可选地或者另外地，如果在与(a)用低浓度突变型p53肽脉冲的抗原阴性、适用hla分子阳性的靶细胞或(b)已引入编码突变型p53的核苷酸序列以使靶细胞表达突变的p53的抗原阴性、适用hla分子阳性的靶细胞共同培养时，通过elispot检测到表达tcr的t细胞的的至少两倍的斑点，则tcr可被视为对突变型p53具有“抗原特异性”，其中上述斑点数是与通过elispot检测的与相同靶细胞共培养的阴性对照t细胞的斑点数相比。肽和阴性对照的浓度可如本文结合关于本发明的其它方面所述。可选地或者另外地，如果对于与以下靶细胞共培养的表达tcr的t细胞而言通过elispot检测到大于约50个斑点，则tcr可被视为对突变型p53具有“抗原特异性”:(a)用低浓度突变型p53肽脉冲的抗原阴性、适用hla分子阳性的靶细胞或(b)已将编码突变型p53的核苷酸序列引入以便靶细胞表达突变型p53的抗原阴性、适用hla分子阳性的靶细胞，。肽的浓度可如本文关于本发明的其它方面所述。可选地或者另外地，如果表达tcr的t细胞上调4-1bb和ox40的一种或两种表达(例如在用表达突变p53的靶细胞刺激后通过流式细胞术测量)，则tcr可被视为对突变p53具有“抗原特异性”。本发明提供包含两个多肽(即多肽链)的tcr，例如tcr的alpha(α)链、tcr的beta(β)链、tcr的gamma(γ)链、tcr的delta(δ)链或其组合。本发明tcr的多肽可包含任何氨基酸序列，前提是tcr对突变的p53具有抗原特异性。在本发明的一个实施方案中，tcr包含两条多肽链，每条多肽链均包含包含tcr的互补决定区(cdr)1、cdr2及cdr3的可变区。在本发明的一个实施方案中，tcr包含第一多肽链和第二多肽链，第一多肽链包含α链cdr1(cdr1α)、α链cdr2(cdr2α)和α链cdr3(cdr3α)的，以及第二多肽链包含β链cdr1(cdr1β)、β链cdr2(cdr2β)和β链cdr3(cdr3β)。在本发明的一个实施方案中，tcr包含以下氨基酸序列:(1)所有seqidno:27-32；(2)所有seqidno:37-42；(3)所有seqidno:47-52；(4)所有seqidno:57-62；(5)所有seqidno:67-72；(6)所有seqidno:77-82；(7)所有seqidno:87-92；(8)所有seqidno:97-102；(9)所有seqidno:107-112；(10)所有seqidno:117-122；(11)所有seqidno:127-132；(12)所有seqidno:137-142；(13)所有seqidno:147-152；(14)所有seqidno:157-162；(15)所有seqidno:167-172；(16)所有seqidno:177-182；(17)所有seqidno:187-192；(18)所有seqidno:197-202；(19)所有seqidno:207-212；(20)所有seqidno:217-222；(21)所有seqidno:227-232；(22)所有seqidno:237-242；(23)所有seqidno:247-252；(24)所有seqidno:257-262；(25)所有seqidno:267-272；(26)所有seqidno:277-282；(27)所有seqidno:287-292；(28)所有seqidno:297-302；(29)所有seqidno:307-312；(30)所有seqidno:317-322；(31)所有seqidno:327-332；(32)所有seqidno:337-342；(33)所有seqidno:347-352；(34)所有seqidno:357-362；(35)所有seqidno:367-372；(36)所有seqidno:377-382；(37)所有seqidno:387-392；(38)所有seqidno:397-402；(39)所有seqidno:407-412；(40)所有seqidno:417-422；(41)所有seqidno:427-432；(42)所有seqidno:437-442；(43)所有seqidno:447-452；(44)所有seqidno:457-462；(45)所有seqidno:467-472；(46)所有seqidno:477-482；或(47)所有seqidno:487-492。本段中前述47个氨基酸序列集合中的每一个列出了对突变人p53具有抗原特异性的47个不同tcr的每一个的6个cdr区域。每一集合中的6个氨基酸序列分别对应于tcr的cdr1α、cdr2α、cdr3α、cdr1β、cdr2β和cdr3β。在本发明的一个实施方案中，tcr包含α链可变区氨基酸序列及β链可变区氨基酸序列，其共同包含上述cdr集合之一。在这方面，tcr可包含以下氨基酸序列:(1)seqidno:33-34两者；(2)seqidno:43-44两者；(3)seqidno:53-54两者；(4)seqidno:63-64两者；(5)seqidno:73-74两者；(6)seqidno:83-84两者；(7)seqidno:93-94两者；(8)seqidno:103-104两者；(9)seqidno:113-114两者；(10)seqidno:123-124两者；(11)seqidno:133-134两者；(12)seqidno:143-144两者；(13)seqidno:153-154两者；(14)seqidno:163-164两者；(15)seqidno:173-174两者；(16)seqidno:183-184两者；(17)seqidno:193-194两者；(18)seqidno:203-204两者；(19)seqidno:213-214两者；(20)seqidno:223-224两者；(21)seqidno:233-234两者；(22)seqidno:243-244两者；(23)seqidno:253-254两者；(24)seqidno:263-264两者；(25)seqidno:273-274两者；(26)seqidno:283-284两者；(27)seqidno:293-294两者；(28)seqidno:303-304两者；(29)seqidno:313-314两者；(30)seqidno:323-324两者；(31)seqidno:333-334两者；(32)seqidno:343-344两者；(33)seqidno:353-354；(34)seqidno:363-364；(35)seqidno:373-374两者；(36)seqidno:383-384两者；(37)seqidno:393-394两者；(38)seqidno:403-404两者；(39)seqidno:413-414两者；(40)seqidno:423-424两者；(41)seqidno:433-434两者；(42)seqidno:443-444两者；(43)seqidno:453-454两者；(44)seqidno:463-464两者；(45)seqidno:473-474两者；(46)seqidno:483-484两者；或(47)seqidno:493-494两者。本段中的前述47个氨基酸序列集合中的每一个列出了对突变人p53具有抗原特异性的47个不同tcr的每一个的两个可变区域。每个集合中的两个氨基酸序列分别对应于tcr的α链的可变区和β链的可变区。本发明tcr可进一步包含恒定区。恒定区可衍生自任何适宜物种(例如人或小鼠)。在本发明的一个实施方案中，tcr进一步包含鼠恒定区。如本文所使用，当提及本文所述的tcr或tcr的任何组分时(例如，互补决定区(cdr)、可变区、恒定区、α链和/或β链)，术语“鼠”或“人”意指分别衍生自小鼠或人的tcr(或其组分)，即，分别起源自或曾经由小鼠t细胞或人t细胞表达的tcr。在本发明的一个实施方案中，tcr可包含鼠α链恒定区及鼠β链恒定区。鼠α链恒定区可是被修饰的或未被修饰的。如例如us2017/0145070中所述，修饰的鼠α链恒定区可以是例如半胱氨酸取代的、lvl修饰的、或同时半胱氨酸取代和lvl修饰的。鼠β链恒定区可以是修饰的或未修饰的。修饰的鼠β链恒定区可以是例如半胱氨酸取代的，如例如us2017/0145070中所述。在本发明的一个实施方案中，tcr包含半胱氨酸取代的、lvl修饰的鼠α链恒定区，其包含seqidno:23或24的氨基酸序列。在本发明的一个实施方案中，tcr包含包含seqidno:25的氨基酸序列的经半胱氨酸取代的鼠β链恒定区。在本发明的一个实施方案中，本发明tcr可包含tcr的α链及tcr的β链。tcr的α链可包含α链的可变区和α链的恒定区。这种类型的α链可与tcr的任何β链配对。β链可包含β链的可变区及β链的恒定区。在本发明的一个实施方案中，tcr包含以下氨基酸序列:(1)seqidno:35-36两者；(2)seqidno:45-46两者；(3)seqidno:55-56两者；(4)seqidno:65-66两者；(5)seqidno:75-76两者；(6)seqidno:85-86两者；(7)seqidno:95-96两者；(8)seqidno:105-106两者；(9)seqidno:115-116两者；(10)seqidno:125-126两者；(11)seqidno:135-136两者；(12)seqidno:145-146两者；(13)seqidno:155-156两者；(14)seqidno:165-166两者；(15)seqidno:175-176两者；(16)seqidno:185-186两者；(17)seqidno:195-196两者；(18)seqidno:205-206两者；(19)seqidno:215-216两者；(20)seqidno:225-226两者；(21)seqidno:235-236两者；(22)seqidno:245-246两者；(23)seqidno:255-256两者；(24)seqidno:265-266两者；(25)seqidno:275-276两者；(26)seqidno:285-286两者；(27)seqidno:295-296两者；(28)seqidno:305-306两者；(29)seqidno:315-316两者；(30)seqidno:325-326两者；(31)seqidno:335-336两者；(32)seqidno:345-346两者；(33)seqidno:355-356两者；(34)seqidno:365-366两者；(35)seqidno:375-376两者；(36)seqidno:385-386两者；(37)seqidno:395-396两者；(38)seqidno:405-406两者；(39)seqidno:415-416两者；(40)seqidno:425-426两者；(41)seqidno:435-436两者；(42)seqidno:445-446两者；(43)seqidno:455-456两者；(44)seqidno:465-466两者；(45)seqidno:475-476两者；(46)seqidno:485-486两者；或(47)seqidno:495-496两者本段中的上述47个氨基酸序列集合中的每一个列出了对突变人p53具有抗原特异性的47个不同tcr的每一个的α链和β链。每个集合中的两个氨基酸序列分别对应于tcr的α链和β链。本发明范围包括本文所述的本发明tcr的功能性变体。如本文所使用的术语“功能性变体”是指与母体tcr、多肽或蛋白质具有实质性或显著序列同一性或相似性的tcr、多肽或蛋白质，该功能性变体保留其为变体的tcr、多肽或蛋白质的生物活性。例如，功能性变体包括本文所述的tcr、多肽或蛋白质(母体tcr、多肽或蛋白质)的那些变体，其保留与母体tcr、多肽或蛋白质具有相似程度、相同程度或更高程度的特异性结合突变p53的能力，所述母体tcr对所述突变p53具有抗原特异性、或所述母体多肽或蛋白质特异性结合所述突变p53。就母体tcr、多肽或蛋白质而言，功能性变体可以例如在氨基酸序列中分别与母体tcr、多肽或蛋白质至少约30％、至少约50％、至少约75％、至少约80％、至少约90％、至少约95％、至少约96％、至少约97％、至少约98％、至少约99％或更多相同。例如，功能性变体可包含具有至少一个保守氨基酸取代的母体tcr、多肽或蛋白质的氨基酸序列。保守氨基酸取代为本领域已知，并且包括氨基酸取代，其中具有某些物理和/或化学性质的一个氨基酸交换为具有相同化学或物理性质的另一个氨基酸。例如，保守氨基酸取代可为酸性氨基酸取代另一酸性氨基酸(例如，asp或glu)，具有非极性侧链的氨基酸取代另一具有非极性侧链的氨基酸(例如，ala、gly、val、ile、leu、met、phe、pro、trp、val、等)，碱性氨基酸被另一种碱性氨基酸(lys、arg等)取代，具有极性侧链的氨基酸取代另一具有极性侧链的氨基酸(asn、cys、gln、ser、thr、tyr等)，等等。可选地或者另外地，功能性变体可包含具有至少一个非保守氨基酸取代的母体tcr、多肽或蛋白质的氨基酸序列。在这种情况下，优选非保守氨基酸取代不干扰或抑制功能性变体的生物活性。优选地，非保守氨基酸取代增强功能性变体的生物活性，使得与母体tcr、多肽或蛋白质相比，功能性变体的生物活性增加。tcr、多肽或蛋白质基本上可由本文所述的指定氨基酸序列或序列组成，使得tcr、多肽或蛋白质的其他组分，例如其他氨基酸，不会实质性地改变tcr、多肽或蛋白质的生物活性。本发明还提供包含本文所述的任一tcr的功能部分的多肽。如本文所使用的术语“多肽”包括寡肽并且是指通过一个或多个肽键连接的氨基酸的单链。关于本发明的多肽，功能部分可以是包含为tcr的一部分的tcr的邻接氨基酸的任何部分，前提是功能部分特异性地结合到突变型p53。当在提及tcr时使用时，术语“功能部分”是指本发明的tcr的任何部分或片段，该部分或片段保留其为其一部分的tcr的生物活性(母体tcr)。例如，功能部分包含tcr的那些部分，其保留与突变p53特异性结合的能力(例如，以适用的hla分子依赖性方式)，或检测、治疗或预防癌症的能力，程度与母体tcr相似、相同或更高。就母体tcr而言，功能部分可包含例如母体tcr的约10％、约25％、约30％、约50％、约68％、约80％、约90％、约95％或更多。上述功能部分可包含该部分的氨基末端或羧基末端或两末端的额外氨基酸，所述额外氨基酸不存在于母体tcr的氨基酸序列中。理想的是，所述额外氨基酸不会干扰功能部分的生物功能，例如，特异性结合突变的p53；和/或能够检测癌症、治疗或预防癌症等。更理想的是，与母体tcr的生物活性相比，所述额外氨基酸增强了生物活性。所述多肽可包含本发明tcr的α链和β链之一或两者的功能部分，例如包含本发明tcr的α链和/或β链的可变区的cdr1、cdr2和cdr3中的一个或多个的功能部分。在本发明的一个实施方案中，多肽可包含的功能部分包含以下氨基酸序列:(1)seqidno:27-32的全部；(2)seqidno:37-42的全部；(3)seqidno:47-52的全部；(4)seqidno:57-62的全部；(5)seqidno:67-72的全部；(6)seqidno:77-82的全部；(7)seqidno:87-92的全部；(8)seqidno:97-102的全部；(9)seqidno:107-112的全部；(10)seqidno:117-122的全部；(11)seqidno:127-132的全部；(12)seqidno:137-142的全部；(13)seqidno:147-152的全部；(14)seqidno:157-162的全部；(15)seqidno:167-172的全部；(16)seqidno:177-182的全部；(17)seqidno:187-192的全部；(18)seqidno:197-202的全部；(19)seqidno:207-212的全部；(20)seqidno:217-222的全部；(21)seqidno:227-232的全部；(22)所有seqidno:237-242的全部；(23)seqidno:247-252的全部；(24)seqidno:257-262的全部；(25)seqidno:267-272的全部；(26)seqidno:277-282的全部；(27)seqidno:287-292的全部；(28)seqidno:297-302的全部；(29)seqidno:307-312的全部；(30)seqidno:317-322的全部；(31)seqidno:327-332的全部；(32)seqidno:337-342的全部；(33)seqidno:347-352的全部；(34)seqidno:357-362的全部；(35)seqidno:367-372的全部；(36)seqidno:377-382的全部；(37)seqidno:387-392的全部；(38)seqidno:397-402的全部；(39)所有seqidno:407-412的全部；(40)seqidno:417-422的全部；(41)seqidno:427-432的全部；(42)seqidno:437-442的全部；(43)seqidno:447-452的全部；(44)seqidno:457-462的全部；(45)seqidno:467-472的全部；(46)seqidno:477-482的全部；或(47)seqidno:487-492的全部。在本发明的一个实施方案中，本发明多肽可包含(例如)本发明tcr的可变区，其包含上述cdr区之组合。在这方面，多肽可包含以下氨基酸序列:(1)seqidno:33-34两者；(2)seqidno:43-44两者；(3)seqidno:53-54两者；(4)seqidno:63-64两者；(5)seqidno:73-74两者；(6)seqidno:83-84两者；(7)seqidno:93-94两者；(8)seqidno:103-104两者；(9)seqidno:113-114两者；(10)seqidno:123-124两者；(11)seqidno:133-134两者；(12)seqidno:143-144两者；(13)seqidno:153-154两者；(14)seqidno:163-164两者；(15)seqidno:173-174两者；(16)seqidno:183-184两者；(17)seqidno:193-194两者；(18)seqidno:203-204两者；(19)seqidno:213-214两者；(20)seqidno:223-224两者；(21)seqidno:233-234两者；(22)seqidno:243-244两者；(23)seqidno:253-254两者；(24)seqidno:263-264两者；(25)seqidno:273-274两者；(26)seqidno:283-284两者；(27)seqidno:293-294两者；(28)seqidno:303-304两者；(29)seqidno:313-314两者；(30)seqidno:323-324两者；(31)seqidno:333-334两者；(32)seqidno:343-344两者；(33)seqidno:353-354两者；(34)seqidno:363-364两者；(35)seqidno:373-374两者；(36)seqidno:383-384两者；(37)seqidno:393-394两者；(38)seqidno:403-404两者；(39)seqidno:413-414两者；(40)seqidno:423-424；(41)seqidno:433-434两者；(42)seqidno:443-444两者；(43)seqidno:453-454两者；(44)seqidno:463-464两者；(45)seqidno:473-474两者；(46)seqidno:483-484两者；或(47)seqidno:493-494两者。在本发明的一个实施方案中，本发明多肽可进一步包含上述本发明tcr的恒定区。在这方面，多肽可包含(i)seqidno:23-25之一或(ii)seqidno:25和seqidno:23-24之一的氨基酸序列。在本发明的一个实施方案中，本发明多肽可包含本发明tcr的α链及β链。在这方面，多肽可包含以下氨基酸序列:(1)seqidno:35-36两者；(2)seqidno:45-46两者；(3)seqidno:55-56两者；(4)seqidno:65-66两者；(5)seqidno:75-76两者；(6)seqidno:85-86两者；(7)seqidno:95-96两者；(8)seqidno:105-106两者；(9)seqidno:115-116两者；(10)seqidno:125-126两者；(11)seqidno:135-136两者；(12)seqidno:145-146两者；(13)seqidno:155-156两者；(14)seqidno:165-166两者；(15)seqidno:175-176两者；(16)seqidno:185-186两者；(17)seqidno:195-196两者；(18)seqidno:205-206两者；(19)seqidno:215-216两者；(20)seqidno:225-226两者；(21)seqidno:235-236两者；(22)seqidno:245-246两者；(23)seqidno:255-256两者；(24)seqidno:265-266；(25)seqidno:275-276两者；(26)seqidno:285-286两者；(27)seqidno:295-296两者；(28)seqidno:305-306两者；(29)seqidno:315-316两者；(30)seqidno:325-326两者；(31)seqidno:335-336两者；(32)seqidno:345-346两者；(33)seqidno:355-356两者；(34)seqidno:365-366两者；(35)seqidno:375-376两者；(36)seqidno:385-386两者；(37)seqidno:395-396两者；(38)seqidno:405-406两者；(39)seqidno:415-416两者；(40)seqidno:425-426两者；(41)seqidno:435-436两者；(42)seqidno:445-446两者；(43)seqidno:455-456；(44)seqidno:465-466两者；(45)seqidno:475-476两者；(46)seqidno:485-486两者；或(47)seqidno:495-496两者。本发明进一步提供包含本文所述的至少一种多肽的蛋白质。“蛋白质”意指包含一个或多个多肽链的分子。在一个实施方案中，本发明的蛋白质可包含:(1)包含seqidno:27-29全部的氨基酸序列的第一多肽链及包含所有seqidno:30-32的氨基酸序列的第二多肽链；(2)包含seqidno:37-39全部的氨基酸序列的第一多肽链和包含seqidno:40-42全部的氨基酸序列的第二多肽链；(3)包含seqidno:47-49全部的氨基酸序列的第一多肽链和包含seqidno:50-52全部的氨基酸序列的第二多肽链；(4)包含seqidno:57-59全部的氨基酸序列的第一多肽链和包含seqidno:60-62全部的氨基酸序列的第二多肽链；(5)包含seqidno:67-69全部的氨基酸序列的第一多肽链，和包含seqidno:70-72全部的氨基酸序列的第二多肽链；(6)包含seqidno:77-79全部的氨基酸序列的第一多肽链和包含seqidno:80-82全部的氨基酸序列的第二多肽链；(7)包含seqidno:87-89全部的氨基酸序列的第一多肽链和包含seqidno:90-92全部的氨基酸序列的第二多肽链；(8)包含seqidno:97-99全部的氨基酸序列的第一多肽链和包含seqidno:100-102全部的氨基酸序列的第二多肽链；(9)包含seqidno:107-109全部的氨基酸序列的第一多肽链和包含seqidno:110-112全部的氨基酸序列的第二多肽链；(10)包含seqidno:117-119全部的氨基酸序列的第一多肽链和包含seqidno:120-122全部的氨基酸序列的第二多肽链；(11)包含seqidno:127-129全部的氨基酸序列的第一多肽链和包含seqidno:130-132全部的氨基酸序列的第二多肽链；(12)包含seqidno:137-139全部的氨基酸序列的第一多肽链和包含seqidno:140-142全部的氨基酸序列的第二多肽链；(13)第一多肽链，其包含seqidno:147-149全部的氨基酸序列，和第二多肽链，其包含seqidno:150-152全部的氨基酸序列；(14)第一多肽链，其包含seqidno:157-159全部的氨基酸序列，和第二多肽链，其包含seqidno:160-162全部的氨基酸序列；(15)包含seqidno:167-169全部的氨基酸序列的第一多肽链，和包含seqidno:170-172全部的氨基酸序列的第二多肽链；(16)包含seqidno:177-179全部的氨基酸序列的第一多肽链，和包含seqidno:180-182全部的氨基酸序列的第二多肽链；(17)第一多肽链，其包含seqidno:187-189全部的氨基酸序列，和第二多肽链，其包含seqidno:190-192全部的氨基酸序列；(18)第一多肽链，其包含seqidno:197-199全部的氨基酸序列，和第二多肽链，其包含seqidno:200-202全部的氨基酸序列；(19)第一多肽链，其包含seqidno:207-209全部的氨基酸序列，和第二多肽链，其包含seqidno:210-212全部的氨基酸序列；(20)第一多肽链，其包含seqidno:217-219全部的氨基酸序列，和第二多肽链，其包含seqidno:220-222全部的氨基酸序列；(21)包含seqidno:227-229全部的氨基酸序列的第一多肽链，和包含seqidno:230-232全部的氨基酸序列的第二多肽链；(22)包含seqidno:237-239全部的氨基酸序列的第一多肽链，和包含seqidno:240-242全部的氨基酸序列的第二多肽链；(23)第一多肽链，其包含seqidno:247-249全部的氨基酸序列，和第二多肽链，其包含seqidno:250-252全部的氨基酸序列；(24)第一多肽链，其包含seqidno:257-259全部的氨基酸序列，和第二多肽链，其包含seqidno:260-262全部的氨基酸序列；(25)第一多肽链，其包含seqidno:267-269全部的氨基酸序列，和第二多肽链，其包含seqidno:270-272全部的氨基酸序列；(26)第一多肽链，其包含seqidno:277-279全部的氨基酸序列，和第二多肽链，其包含seqidno:280-282全部的氨基酸序列；(27)第一多肽链，其包含seqidno:287-289全部的氨基酸序列，和第二多肽链，其包含seqidno:290-292全部的氨基酸序列；(28)第一多肽链，其包含seqidno:297-299全部的氨基酸序列，和第二多肽链，其包含seqidno:300-302全部的氨基酸序列；(29)第一多肽链，其包含seqidno:307-309全部的氨基酸序列，和第二多肽链，其包含seqidno:310-312全部的氨基酸序列；(30)包含seqidno:317-319全部的氨基酸序列的第一多肽链，和包含seqidno:320-322全部的氨基酸序列的第二多肽链；(31)包含seqidno:327-329全部的氨基酸序列的第一多肽链，和包含seqidno:330-332全部的氨基酸序列的第二多肽链；(32)第一多肽链，其包含seqidno:337-339全部的氨基酸序列，和第二多肽链，其包含seqidno:340-342全部的氨基酸序列；(33)第一多肽链，其包含seqidno:347-349全部的氨基酸序列，和第二多肽链，其包含seqidno:350-352全部的氨基酸序列；(34)包含seqidno:357-359全部的氨基酸序列的第一多肽链，和包含seqidno:360-362全部的氨基酸序列的第二多肽链；(35)包含seqidno:367-369全部的氨基酸序列的第一多肽链，和包含seqidno:370-372全部的氨基酸序列的第二多肽链；(36)第一多肽链，其包含seqidno:377-379全部的氨基酸序列，和第二多肽链，其包含seqidno:380-382全部的氨基酸序列；(37)第一多肽链，其包含seqidno:387-389全部的氨基酸序列和第二多肽链，其包含seqidno:390-392全部的氨基酸序列；(38)包含seqidno:397-399全部的氨基酸序列的第一多肽链，和包含seqidno:400-402全部的氨基酸序列的第二多肽链；(39)包含seqidno:407-409全部的氨基酸序列的第一多肽链和包含seqidno:410-412全部的氨基酸序列的第二多肽链；(40)包含seqidno:417-419全部的氨基酸序列的第一多肽链和包含seqidno:420-422全部的氨基酸序列的第二多肽链；(41)第一多肽链，其包含seqidno:427-429全部的氨基酸序列，和第二多肽链，其包含seqidno:430-432全部的氨基酸序列；(42)第一多肽链，其包含seqidno:437-439全部的氨基酸序列，和第二多肽链，其包含seqidno:440-442全部的氨基酸序列；(43)第一多肽链，其包含seqidno:447-449全部的氨基酸序列，和第二多肽链，其包含seqidno:450-452全部的氨基酸序列；(44)包含seqidno:457-459全部的氨基酸序列的第一多肽链，和包含seqidno:460-462全部的氨基酸序列的第二多肽链；(45)第一多肽链，其包含seqidno:467-469全部的氨基酸序列，和第二多肽链，其包含seqidno:470-472全部的氨基酸序列；(46)第一多肽链，其包含seqidno:477-479全部的氨基酸序列，和第二多肽链，其包含seqidno:480-482全部的氨基酸序列；或(47)包含seqidno:487-489全部的氨基酸序列的第一多肽链，和包含seqidno:490-492全部的氨基酸序列的第二多肽链。在本发明的一个实施方案中，蛋白质包含:(1)包含seqidno:33的氨基酸序列的第一多肽链及包含seqidno:34的氨基酸序列的第二多肽链；(2)包含seqidno:43的氨基酸序列的第一多肽链和包含seqidno:44的氨基酸序列的第二多肽链；(3)包含seqidno:53的氨基酸序列的第一多肽链和包含seqidno:54的氨基酸序列的第二多肽链；(4)包含seqidno:63的氨基酸序列的第一多肽链和包含seqidno:64的氨基酸序列的第二多肽链；(5)包含seqidno:73的氨基酸序列的第一多肽链和包含seqidno:74的氨基酸序列的第二多肽链；(6)包含seqidno:83的氨基酸序列的第一多肽链和包含seqidno:84的氨基酸序列的第二多肽链；(7)包含seqidno:93的氨基酸序列的第一多肽链和包含seqidno:94的氨基酸序列的第二多肽链；(8)包含seqidno:103的氨基酸序列的第一多肽链和包含seqidno:104的氨基酸序列的第二多肽链；(9)包含seqidno:113的氨基酸序列的第一多肽链和包含seqidno:114的氨基酸序列的第二多肽链；(10)包含seqidno:123的氨基酸序列的第一多肽链和包含seqidno:124的氨基酸序列的第二多肽链；(11)包含seqidno:133的氨基酸序列的第一多肽链和包含seqidno:134的氨基酸序列的第二多肽链；(12)包含seqidno:143的氨基酸序列的第一多肽链和包含seqidno:144的氨基酸序列的第二多肽链；(13)包含seqidno:153的氨基酸序列的第一多肽链和包含seqidno:154的氨基酸序列的第二多肽链；(14)包含seqidno:163的氨基酸序列的第一多肽链和包含seqidno:164的氨基酸序列的第二多肽链；(15)包含seqidno:173的氨基酸序列的第一多肽链和包含seqidno:174的氨基酸序列的第二多肽链；(16)包含seqidno:183的氨基酸序列的第一多肽链和包含seqidno:184的氨基酸序列的第二多肽链；(17)包含seqidno:193的氨基酸序列的第一多肽链和包含seqidno:194的氨基酸序列的第二多肽链；(18)包含seqidno:203的氨基酸序列的第一多肽链和包含seqidno:204的氨基酸序列的第二多肽链；(19)包含seqidno:213的氨基酸序列的第一多肽链和包含seqidno:214的氨基酸序列的第二多肽链；(20)包含seqidno:223的氨基酸序列的第一多肽链和包含seqidno:224的氨基酸序列的第二多肽链；(21)包含seqidno:233的氨基酸序列的第一多肽链和包含seqidno:234的氨基酸序列的第二多肽链；(22)包含seqidno:243的氨基酸序列的第一多肽链和包含seqidno:244的氨基酸序列的第二多肽链；(23)包含seqidno:253的氨基酸序列的第一多肽链和包含seqidno:254的氨基酸序列的第二多肽链；(24)包含seqidno:263的氨基酸序列的第一多肽链和包含seqidno:264的氨基酸序列的第二多肽链；(25)包含seqidno:273的氨基酸序列的第一多肽链和包含seqidno:274的氨基酸序列的第二多肽链；(26)包含seqidno:283的氨基酸序列的第一多肽链和包含seqidno:284的氨基酸序列的第二多肽链；(27)包含seqidno:293的氨基酸序列的第一多肽链和包含seqidno:294的氨基酸序列的第二多肽链；(28)包含seqidno:303的氨基酸序列的第一多肽链和包含seqidno:304的氨基酸序列的第二多肽链；(29)包含seqidno:313的氨基酸序列的第一多肽链和包含seqidno:314的氨基酸序列的第二多肽链；(30)包含seqidno:323的氨基酸序列的第一多肽链和包含seqidno:324的氨基酸序列的第二多肽链；(31)包含seqidno:333的氨基酸序列的第一多肽链和包含seqidno:334的氨基酸序列的第二多肽链；(32)包含seqidno:343的氨基酸序列的第一多肽链和包含seqidno:344的氨基酸序列的第二多肽链；(33)包含seqidno:353的氨基酸序列的第一多肽链和包含seqidno:354的氨基酸序列的第二多肽链；(34)包含seqidno:363的氨基酸序列的第一多肽链和包含seqidno:364的氨基酸序列的第二多肽链；(35)包含seqidno:373的氨基酸序列的第一多肽链和包含seqidno:374的氨基酸序列的第二多肽链；(36)包含seqidno:383的氨基酸序列的第一多肽链和包含seqidno:384的氨基酸序列的第二多肽链；(37)包含seqidno:393的氨基酸序列的第一多肽链和包含seqidno:394的氨基酸序列的第二多肽链；(38)包含seqidno:403的氨基酸序列的第一多肽链和包含seqidno:404的氨基酸序列的第二多肽链；(39)包含seqidno:413的氨基酸序列的第一多肽链和包含seqidno:414的氨基酸序列的第二多肽链；(40)包含seqidno:423的氨基酸序列的第一多肽链和包含seqidno:424的氨基酸序列的第二多肽链；(41)包含seqidno:433的氨基酸序列的第一多肽链和包含seqidno:434的氨基酸序列的第二多肽链；(42)包含seqidno:443的氨基酸序列的第一多肽链和包含seqidno:444的氨基酸序列的第二多肽链；(43)包含seqidno:453的氨基酸序列的第一多肽链和包含seqidno:454的氨基酸序列的第二多肽链；(44)包含seqidno:463的氨基酸序列的第一多肽链和包含seqidno:464的氨基酸序列的第二多肽链；(45)包含seqidno:473的氨基酸序列的第一多肽链和包含seqidno:474的氨基酸序列的第二多肽链；(46)包含seqidno:483的氨基酸序列的第一多肽链和包含seqidno:484的氨基酸序列的第二多肽链；或(47)包含seqidno:493的氨基酸序列的第一多肽链和包含seqidno:494的氨基酸序列的第二多肽链。在本发明的一个实施方案中，蛋白质包含:(1)包含seqidno:35的氨基酸序列的第一多肽链及包含seqidno:36的氨基酸序列的第二多肽链；(2)包含seqidno:45的氨基酸序列的第一多肽链和包含seqidno:46的氨基酸序列的第二多肽链；(3)包含seqidno:55的氨基酸序列的第一多肽链和包含seqidno:56的氨基酸序列的第二多肽链；(4)包含seqidno:65的氨基酸序列的第一多肽链和包含seqidno:66的氨基酸序列的第二多肽链；(5)包含seqidno:75的氨基酸序列的第一多肽链和包含seqidno:76的氨基酸序列的第二多肽链；(6)包含seqidno:85的氨基酸序列的第一多肽链和包含seqidno:86的氨基酸序列的第二多肽链；(7)包含seqidno:95的氨基酸序列的第一多肽链和包含seqidno:96的氨基酸序列的第二多肽链；(8)包含seqidno:105的氨基酸序列的第一多肽链和包含seqidno:106的氨基酸序列的第二多肽链；(9)包含seqidno:115的氨基酸序列的第一多肽链和包含seqidno:116的氨基酸序列的第二多肽链；(10)包含seqidno:125的氨基酸序列的第一多肽链和包含seqidno:126的氨基酸序列的第二多肽链；(11)包含seqidno:135的氨基酸序列的第一多肽链和包含seqidno:136的氨基酸序列的第二多肽链；(12)包含seqidno:145的氨基酸序列的第一多肽链和包含seqidno:146的氨基酸序列的第二多肽链；(13)包含seqidno:155的氨基酸序列的第一多肽链和包含seqidno:156的氨基酸序列的第二多肽链；(14)包含seqidno:165的氨基酸序列的第一多肽链和包含seqidno:166的氨基酸序列的第二多肽链；(15)包含seqidno:175的氨基酸序列的第一多肽链和包含seqidno:176的氨基酸序列的第二多肽链；(16)包含seqidno:185的氨基酸序列的第一多肽链和包含seqidno:186的氨基酸序列的第二多肽链；(17)包含seqidno:195的氨基酸序列的第一多肽链和包含seqidno:196的氨基酸序列的第二多肽链；(18)包含seqidno:205的氨基酸序列的第一多肽链和包含seqidno:206的氨基酸序列的第二多肽链；(19)包含seqidno:215的氨基酸序列的第一多肽链和包含seqidno:216的氨基酸序列的第二多肽链；(20)包含seqidno:225的氨基酸序列的第一多肽链和包含seqidno:226的氨基酸序列的第二多肽链；(21)包含seqidno:235的氨基酸序列的第一多肽链和包含seqidno:236的氨基酸序列的第二多肽链；(22)包含seqidno:245的氨基酸序列的第一多肽链和包含seqidno:246的氨基酸序列的第二多肽链；(23)包含seqidno:255的氨基酸序列的第一多肽链和包含seqidno:256的氨基酸序列的第二多肽链；(24)包含seqidno:265的氨基酸序列的第一多肽链和包含seqidno:266的氨基酸序列的第二多肽链；(25)包含seqidno:275的氨基酸序列的第一多肽链和包含seqidno:276的氨基酸序列的第二多肽链；(26)包含seqidno:285的氨基酸序列的第一多肽链和包含seqidno:286的氨基酸序列的第二多肽链；(27)包含seqidno:295的氨基酸序列的第一多肽链和包含seqidno:296的氨基酸序列的第二多肽链；(28)包含seqidno:305的氨基酸序列的第一多肽链和包含seqidno:306的氨基酸序列的第二多肽链；(29)包含seqidno:315的氨基酸序列的第一多肽链和包含seqidno:316的氨基酸序列的第二多肽链；(30)包含seqidno:325的氨基酸序列的第一多肽链和包含seqidno:326的氨基酸序列的第二多肽链；(31)包含seqidno:335的氨基酸序列的第一多肽链和包含seqidno:336的氨基酸序列的第二多肽链；(32)包含seqidno:345的氨基酸序列的第一多肽链和包含seqidno:346的氨基酸序列的第二多肽链；(33)包含seqidno:355的氨基酸序列的第一多肽链和包含seqidno:356的氨基酸序列的第二多肽链；(34)包含seqidno:365的氨基酸序列的第一多肽链和包含seqidno:366的氨基酸序列的第二多肽链；(35)包含seqidno:375的氨基酸序列的第一多肽链和包含seqidno:376的氨基酸序列的第二多肽链；(36)包含seqidno:385的氨基酸序列的第一多肽链和包含seqidno:386的氨基酸序列的第二多肽链；(37)包含seqidno:395的氨基酸序列的第一多肽链和包含seqidno:396的氨基酸序列的第二多肽链；(38)包含seqidno:405的氨基酸序列的第一多肽链和包含seqidno:406的氨基酸序列的第二多肽链；(39)包含seqidno:415的氨基酸序列的第一多肽链和包含seqidno:416的氨基酸序列的第二多肽链；(40)包含seqidno:425的氨基酸序列的第一多肽链和包含seqidno:426的氨基酸序列的第二多肽链；(41)包含seqidno:435的氨基酸序列的第一多肽链和包含seqidno:436的氨基酸序列的第二多肽链；(42)包含seqidno:445的氨基酸序列的第一多肽链和包含seqidno:446的氨基酸序列的第二多肽链；(43)包含seqidno:455的氨基酸序列的第一多肽链和包含seqidno:456的氨基酸序列的第二多肽链；(44)包含seqidno:465的氨基酸序列的第一多肽链和包含seqidno:466的氨基酸序列的第二多肽链；(45)包含seqidno:475的氨基酸序列的第一多肽链和包含seqidno:476的氨基酸序列的第二多肽链；(46)包含seqidno:485的氨基酸序列的第一多肽链和包含seqidno:486的氨基酸序列的第二多肽链；或(47)包含seqidno:495的氨基酸序列的第一多肽链和包含seqidno:496的氨基酸序列的第二多肽链。本发明的蛋白质可以是tcr。或者，如果蛋白质的第一和/或第二多肽链还包含其他氨基酸序列，例如编码免疫球蛋白或其一部分的氨基酸序列，那么本发明的蛋白质可以是融合蛋白。在这方面，本发明还提供包含本文所述的至少一种本发明多肽以及至少一种其它多肽的融合蛋白。其他多肽可作为融合蛋白的单独多肽存在，或可作为多肽存在，其与本文所述的一种本发明多肽同框(串联)表达。其他多肽可编码任何肽或蛋白质分子或其一部分，包括但不限于免疫球蛋白、cd3、cd4、cd8、mhc分子、cd1分子，例如cd1a、cd1b、cd1c、cd1d等。融合蛋白可包含本发明多肽的一个或多个拷贝和/或其他多肽的一或多个拷贝。例如，融合蛋白可包含本发明多肽和/或其他多肽的1个、2个、3个、4个、5个或更多个拷贝。制备融合蛋白的适宜方法是本领域已知的，且包括(例如)重组方法。在本发明的一些实施方案中，本发明的tcr、多肽和蛋白质可表达为包含连接α链和β链的连接肽的单一蛋白质。在这方面，本发明的tcr、多肽和蛋白质可进一步包含连接子肽。连接肽可有利地促进重组tcr、多肽和/或蛋白在宿主细胞中的表达。连接肽可包含任何适宜的氨基酸序列。例如，连接肽可包含seqidno:26的氨基酸序列。通过宿主细胞表达包括连接肽在内的构建体时，连接肽可能被裂解，从而产生分开的α和β链。在本发明的一个实施方案中，tcr、多肽或蛋白质可包含包含全长α链、全长β链、及位于α与β链之间的连接肽的氨基酸序列。本发明的蛋白质可为重组抗体或其抗原结合部分，其包含本文所述的至少一种本发明的多肽。如本文所用，“重组抗体”是指包含本发明的至少一种多肽和抗体的多肽链或其抗原结合部分的重组(例如，基因工程化的)蛋白。抗体的多肽或其抗原结合部分可为重链、轻链、重链或轻链的可变区或恒定区、单链可变区片段(scfv)或抗体的fc、fab或f(ab)2’片段等。抗体的多肽链或其抗原结合部分可作为重组抗体的单独多肽存在。或者，抗体的多肽链或其抗原结合部分可作为多肽存在，其与本发明的多肽同框(串联)表达。抗体的多肽或其抗原结合部分可以是任何抗体或任何抗体片段的多肽，包括本文所述的任何抗体和抗体片段。本发明的tcr、多肽和蛋白质可具有任何长度，即可包含任何数量的氨基酸，前提是tcr、多肽或蛋白质保持其生物活性，例如特异性结合突变p53的能力、检测哺乳动物中癌症的能力、或治疗或预防哺乳动物中的癌症的能力等。例如，多肽可在约50至约5000个氨基酸长度范围内，例如50、70、75、100、125、150、175、200、300、400、500、600、700、800、900、1000或更多个氨基酸。在这方面，本发明的多肽也包括寡肽。本发明的tcr、多肽和蛋白质可包含合成氨基酸替代一种或多种天然存在的氨基酸。此类合成氨基酸在本领域内是已知的，并且包括例如氨基环己烷羧酸、正亮氨酸、α-氨基正癸酸、高丝氨酸、s-乙酰氨基甲基-半胱氨酸、反式-3-和反式-4-羟基脯氨酸、4-氨基苯丙氨酸、4-硝基苯丙氨酸，4-氯苯丙氨酸、4-羧基苯丙氨酸、β-苯丝氨酸β-羟基苯丙氨酸、苯甘氨酸、α-萘丙氨酸、环己基丙氨酸、环己基甘氨酸、吲哚-2-羧酸、1,2,3,4-四氢异喹啉-3-羧酸、氨基马来酸、氨基马来酸单酰胺，n’-苄基-n’-甲基-赖氨酸，n’，n’-二苄基-赖氨酸，6-羟基赖氨酸，鸟氨酸，α-氨基环戊烷羧酸，α-氨基环己烷羧酸，α-氨基环庚烷羧酸，α-(2-氨基-2-正硼烷)-羧酸，α，β-二氨基丁酸，α，β--二氨基丙酸，同苯丙氨酸和α-叔丁基甘氨酸。本发明的tcr、多肽和蛋白质可被糖基化、酰胺化、羧基化、磷酸化、酯化、n-酰化、例如经二硫键环化、或转化为酸加成盐和/或任选地二聚的或聚合的、或共轭的。本发明的tcr、多肽、和/或蛋白质可藉由本领域已知方法(例如，从头合成)获得。此外，可使用本文所述核酸使用标准重组方法重组产生多肽和蛋白质。例如，参见greenandsambrook,molecularcloning:alaboratorymanual,4thed.,coldspringharborpress,coldspringharbor,ny(2012)。可选地，本文所述的tcr、多肽和/或蛋白质可由诸如synpep(dublin,ca)、peptidetechnologiescorp.(gaithersburg,md)和多肽系统(sandiego,ca)等公司商业化合成。在这方面，本发明tcr、多肽和蛋白质可为合成、重组、分离和/或纯化的。本发明的一个实施方案提供包含编码本文所述的任何tcr、多肽或蛋白质的核苷酸序列的核酸。如本文所使用的“核酸”包括“多核苷酸”、“寡核苷酸”和“核酸分子”，并且通常意指dna或rna的聚合物，其可为单链或双链，其可含有天然、非天然或改变的核苷酸，并且可含有天然、非天然或改变的核苷酸间链接，例如磷酰胺酯链接或硫代磷酸酯链接，而不是未修饰寡核苷酸的核苷酸之间发现的磷酸二酯。在一个实施方案中，核酸包含互补dna(cdna)。通常优选地，核酸不包含任何插入、缺失、倒置和/或取代。然而，在一些实例中，如本文所讨论，其可适用于核酸包含一个或多个插入、删除、倒置和/或取代。优选地，本发明的核酸是重组的。如本文所用，术语“重组”是指(i)通过将天然或合成核酸片段连接到可在活细胞中复制的核酸分子而在活细胞外构建的分子，或(ii)由上文(i)中所述的那些复制产生的分子。出于本文的目的，复制可以是体外复制或体内复制。核酸可使用本领域已知程序基于化学合成和/或酶连接反应构建。例如，参见green和sambrook等人，同上。例如，核酸可使用天然存在的核苷酸或设计用于增加分子的生物稳定性或提高杂交后形成的双链体的物理稳定性的各种修饰的核苷酸(例如，硫代磷酸酯衍生物和吖啶取代的核苷酸)进行化学合成。可用于产生核酸的修饰核苷酸的实例包括但不限于5-氟尿嘧啶、5-溴尿嘧啶、5-氯尿嘧啶、5-碘尿嘧啶、次黄嘌呤、黄嘌呤、4-乙酰胞嘧啶、5-(羧羟甲基)尿嘧啶、5-羧甲基氨基甲基-2-硫脲嘧啶、5-羧甲基氨基甲基尿嘧啶、二氢尿嘧啶、β-d-半乳糖喹啉、肌苷、n6-异戊烯基腺嘌呤、1-甲基鸟嘌呤、1-甲基亚麻苷、2,2-二甲基鸟嘌呤、2-甲基腺嘌呤、2-甲基鸟嘌呤、3-甲基胞嘧啶、5-甲基胞嘧啶、n6-取代的腺嘌呤、7-甲基鸟嘌呤、5-甲基氨基甲基尿嘧啶、5-甲氧氨基甲基-2-硫尿嘧啶、β-d-甘露糖苷、5'-甲氧羧甲基尿嘧啶、5-甲氧基尿嘧啶、2-甲硫基-n6-异戊烯基腺嘌呤、尿嘧啶-5-氧乙酸(v)、wybutoxosine、假性尿嘧啶、queosine、2-硫代胞嘧啶、5-甲基-2-硫尿嘧啶、2-硫尿嘧啶、4-硫尿嘧啶、5-甲基尿嘧啶、尿嘧啶-5-羟乙酸甲酯、3-(3-氨基-3-n-2-羧丙基)尿嘧啶和2,6-二氨基嘌呤。或者，本发明的一种或多种核酸可购自诸如macromolecularresources(fortcollins,co)和synthegen(houston,tx)等公司。在本发明的一个实施方案中，核酸包含编码本文所述的任何tcr、多肽或蛋白质的密码子优化核苷酸序列。在不受任何特定理论或机制约束的情况下，核苷酸序列的密码子优化被认为提高了mrna转录本的翻译效率。核苷酸序列的密码子优化可能涉及将天然密码子替换为编码相同氨基酸的另一密码子，但可通过在细胞内更容易获得的trna进行翻译，从而提高翻译效率。核苷酸序列的优化也可减少干扰翻译的次级mrna结构，从而提高翻译效率。本发明还提供包含与本文所述的任何核酸的核苷酸序列互补的核苷酸序列或在严格条件下与本文所述的任何核酸的核苷酸序列杂交的核苷酸序列的核酸。在严格条件下杂交的核苷酸序列最好在高严格条件下杂交。所述“高严格条件”意指核苷酸序列以比非特异性杂交可更强检测的量特异性地杂交至目标序列(本文所述的任何核酸的核苷酸序列)。高严格性条件包括能够将具有精确互补序列或仅包含几个分散不匹配的多核苷酸与随机序列区分的条件，所述随机序列恰好有几个与核苷酸序列匹配的小区域(例如，3-10个碱基)。这种互补的小区域比14-17碱基或更多碱基的全长互补物更容易熔融，高严格性杂交使它们容易区分。相对较高的严格条件将包括(例如)在约50-70℃的温度下由约0.02-0.1mnacl或其等效物提供的低盐和/或高温条件。此等高严格条件耐受核苷酸序列与模板或靶链之间的极少不匹配(如有的话)，并且尤其适于检测任何本发明tcr的表达。通常可以理解，通过增加甲酰胺的量可以使条件变得更加严格。本发明还提供包含与本文所述的任一核酸具有至少约70％或更多，例如约80％、约90％、约91％、约92％、约93％、约94％、约95％、约96％、约97％、约98％或约99％等同的核苷酸序列的核酸于。在这方面，核酸可以基本上由本文所述的任何核苷酸序列组成。本发明的核酸可并入重组表达载体中。在这方面，本发明提供包含本发明的任何核酸的重组表达载体。在本发明的一个实施方案中，重组表达载体包含编码α链、β链和连接肽的核苷酸序列。出于本文的目的，术语“重组表达载体”意指基因修饰的寡核苷酸或多核苷酸构建体，当构建体包含编码mrna、蛋白质、多肽或肽的核苷酸序列且载体在足以使mrna、蛋白质、多肽或肽在细胞内表达的条件下接触细胞时，所述构建体允许宿主细胞表达rna、蛋白质、多肽或肽。本发明的载体作为整体并非自然存在的。然而，载体的部分可以是自然存在的。本发明的重组表达载体可包含任何类型的核苷酸，包括但不限于dna和rna，其可以是部分由天然来源获得或合成的单链或双链，并且可含有天然、非天然或改变的核苷酸。重组表达载体可包含自然存在的、非自然存在的核苷酸间链接或两种类型的链接。优选地，非自然存在或改变的核苷酸或核苷酸间链接不妨碍载体的转录或复制。本发明的重组表达载体可以是任何合适的重组表达载体，并且可以用于转化或转染任何合适的宿主细胞。合适的载体包括那些设计用于增殖和扩增或用于表达或用于上述两者的载体，例如质粒和病毒。载体可选自转座子/转座酶系列、puc系列(fermentaslifesciences)、pbluescript系列(stratagene，lajolla，ca)、pet系列(novagen，madison，wi)、pgex系列(pharmaciabiotech，uppsala，瑞典)、以及pex系列(clontech,paloalto,ca)。也可使用噬菌体载体，如λgt10、λgt11、λzapii(stratagene)、λembl4和λnm1149。植物表达载体的实例包括pbi01、pbi101.2、pbi101.3、pbi121和pbin19(clontech)。动物表达载体的实例包括peuk-cl、pmam和pmamneo(clontech)。优选地，重组表达载体是转座子载体或病毒载体，例如逆转录病毒载体。本发明的重组表达载体可使用例如green和sambrook等人(同上述)所述的标准重组dna技术制备。可制备环形或线性表达载体的构建体，以包含在原核或真核宿主细胞中发挥功能的复制系统。复制系统可衍生自colel、2μ质粒、λ、sv40、牛乳头瘤病毒等。理想的是，重组表达载体包含调节序列，例如转录和翻译起始和终止密码子，所述调控序列对要将载体引入的宿主细胞类型(例如，细菌、真菌、植物或动物)具有特异性，视情况并考虑载体是基于dna还是基于rna。重组表达载体可包括一个或多个标记基因，可用于选择转化或转染的宿主细胞。标记基因包括杀生物剂耐药性，例如抗生素耐药性、重金属等。在营养缺陷型宿主细胞中互补以提供原养性等。适于本发明表达载体的标记基因包括(例如)新霉素/g418抗性基因、潮霉素抗性基因、组氨酸抗性基因、四环素抗性基因及氨苄西林抗性基因。重组表达载体可包含可操作地连接到编码tcr、多肽或蛋白质的核苷酸序列或与编码tcr、多肽或蛋白质的核苷酸序列互补或杂交的核苷酸序列的天然或非天然启动子。启动子的选择，例如，强、弱、可诱导、组织特异性和发育特异性启动子，在本领域普通技术范围内。类似地，核苷酸序列与启动子的组合也在本领域技术范围内。启动子可为非病毒启动子，例如人延伸因子-1α启动子，或病毒启动子，例如巨细胞病毒(cmv)启动子、sv40启动子、rsv启动子和在鼠干细胞病毒的长末端重复序列中发现的启动子。本发明重组表达载体可设计用于瞬时表达、稳定表达或两者。此外，重组表达载体可用于组成型表达或诱导型表达。此外，可制备重组表达载体以包括自杀基因。如本文所用，术语“自杀基因”是指使表达自杀基因的细胞死亡的基因。自杀基因可以是使基因表达的细胞对药剂(例如药物)敏感的基因，并且当细胞接触或暴露于该药剂时使细胞死亡。自杀基因在本领域是已知的，并且包括(例如)单纯疱疹病毒(hsv)胸苷激酶(tk)基因、胞嘧啶脱氨酶、嘌呤核苷磷酸化酶和硝基还原酶。本发明的另一实施方案进一步提供包含本文所述的任何重组表达载体的宿主细胞。如本文所用，术语“宿主细胞”是指可包含本发明重组表达载体的任何类型的细胞。宿主细胞可以是真核细胞，例如植物、动物、真菌或藻类，或者可以是原核细胞，例如细菌或原生动物。宿主细胞可为培养细胞或原代细胞，即直接从生物体(例如人)中分离的。宿主细胞可以是贴壁细胞或悬浮细胞，即悬浮生长的细胞。合适的宿主细胞是本领域已知的并且包括例如dh5α大肠杆菌细胞、中国仓鼠卵巢细胞、猴vero细胞、cos细胞、hek293细胞等。为了扩增或复制重组表达载体，宿主细胞优选原核细胞，例如dh5细胞。为了产生重组tcr、多肽或蛋白质，宿主细胞优选哺乳动物细胞。最优选地，宿主细胞是人类细胞。虽然宿主细胞可以是任何细胞类型，可以来自任何类型的组织，并且可以是任何发育阶段的细胞，但宿主细胞优选是外周血淋巴细胞(pbl)或外周血单核细胞(pbmc)。更优选地，宿主细胞是t细胞。出于本文的目的，t细胞可以是任何t细胞，例如培养的t细胞，例如原代t细胞；或来自培养的t细胞系的t细胞，例如jurkat、supt1等；或从哺乳动物获得的t细胞。如果从哺乳动物获得，t细胞可以从多种来源获得，包括但不限于血液、骨髓、淋巴结、胸腺或其他组织或液体。t细胞也可以富集或纯化。优选地，该t细胞是人类t细胞。t细胞可以是任何类型的t细胞并且可以是任何发育阶段的t细胞，包括但不限于cd4+/cd8+双阳性t细胞、cd4+辅助t细胞例如th1和th2细胞、cd4+t细胞、cd8+t细胞(例如细胞毒性t细胞)、肿瘤浸润性淋巴细胞(tils)、记忆性t细胞(例如中心记忆性t细胞和效应记忆性t细胞)、初始t细胞等。本发明还提供包含本文所述至少一种宿主细胞的细胞群。细胞群可为异质性群体，其包含包含所述的任何重组表达载体的宿主细胞以及至少一种其他细胞，例如不包含任何重组表达载体的宿主细胞(例如t细胞)或除t细胞以外的细胞，例如b细胞、巨噬细胞、中性粒细胞、红细胞、肝细胞、内皮细胞、上皮细胞、肌肉细胞、脑细胞等。或者，细胞群可以是基本上均质的群体，其中群体主要包含宿主细胞(例如，基本上由其组成)，所述宿主细胞包含重组表达载体。该群体还可以是细胞的克隆群体，其中该群体的所有细胞均为包含重组表达载体的单一宿主细胞的克隆，使得该群体的所有细胞均包含重组表达载体。在本发明的一个实施方案中，细胞群是包含宿主细胞的克隆群，所述宿主细胞包含如本文所述的重组表达载体。在本发明的一个实施方案中，该群体中细胞的数量可快速扩大。t细胞的数量可通过所属领域内已知的任何一种方法来实现扩增，例如，美国专利8,034,334；美国专利8,383,099；美国专利申请公开第2012/0244133号；dudleyetal.,j.immunother.,26:332-42(2003)；andriddelletal.,j.immunol.methods,128:189-201(1990)。在一个实施方案中，通过用okt3抗体、il-2和饲养pbmc(例如，经辐照的同种异体pbmc)培养t细胞来扩大t细胞的数量。本发明的tcr、多肽、蛋白质、核酸、重组表达载体和宿主细胞(包括其群体)可以是分离的和/或纯化的。本文所用术语“分离的”意指已从其自然环境中移除。本文所用术语“纯化的”意指已增加纯度，其中“纯度”是相对术语，且不必解释为绝对纯度。例如，纯度可为至少约50％、至少约60％、至少约70％、至少约80％、至少约90％、至少约95％或可为约100％。本发明tcr、多肽、蛋白质、核酸、重组表达载体和宿主细胞(包括其群体)，其在下文中统称为“本发明tcr材料”，可被配制为组合物，例如药物组合物。在这方面，本发明提供包含本文所述的tcr、多肽、蛋白质、核酸、表达载体和宿主细胞(包括其群体)中的任一种和药学上可接受的载体的药物组合物。含有任一本发明tcr材料的本发明药物组合物可包含一种以上本发明tcr材料，例如多肽和核酸，或两种或多种不同tcr。或者，药物组合物可包含与另一种药物活性剂或药物组合的本发明tcr材料，所述另一种药物活性剂或药物例如化学治疗剂，例如天冬酰胺酶、白消安、卡铂、顺铂、柔红霉素、阿霉素、氟尿嘧啶、吉西他滨、羟基脲、甲氨蝶呤、紫杉醇、利妥昔单抗、长春碱、长春新碱等优选地，该载体是医药上可接受的载体。关于药物组合物，载体可以是常规用于所考虑的特定本发明tcr材料的任何载体。制备可施用组合物的方法为本领域技术人员已知或显而易见且更详细地阐述于例如remington:thescienceandpracticeofpharmacy,22nded.，pharmaceuticalpress(2012)。优选地，医学上可接受的载体是在使用条件下不具有有害副作用或毒性的载体。载体的选择将部分由特定本发明tcr材料以及用于施用与本发明tcr材料的特定方法决定。因此，存在本发明的药物组合物的各种适宜调配物。适宜调配物可包括用于肠外、皮下、静脉内、肌内、动脉内、鞘内、瘤内或腹膜间施用的任何调配物。一种以上途径可用于施用本发明tcr材料，且在某些情况下，特定途径可提供比另一途径更立即且更有效的响应。优选地，通过例如静脉注射施用本发明tcr材料。当本发明tcr材料是表达本发明tcr的宿主细胞时，注射用细胞的药学可接受的载体可包括任何等渗载体，例如生理盐水(约0.90％w/vnacl水溶液、约300mosm/lnacl水溶液或约9.0gnacl/升水)、normosolr电解质溶液(abbott，芝加哥，il)、plasma-lytea(baxter，deerfield，il)、约5％葡萄糖水溶液或林格乳酸盐。在一个实施例中，用所述药学上可接受的载体添加了人血清蛋白。出于本发明的目的，施用的本发明tcr材料的量或剂量(例如，当本发明tcr材料为一个或多个细胞时的细胞数量)应足以在合理的时间范围内对个体或动物产生治疗或预防反应。例如，本发明tcr材料的剂量应足以与癌症抗原(例如突变的p53)结合，或在自施用时起约2小时或更长时间(例如12至24小时或更长时间)内检测、治疗或预防癌症。在某些实施例中，时间周期甚至可能更长。剂量将通过特定本发明tcr材料的有效性和动物(例如人)的状况以及待处理动物(例如人)的体重确定。确定给药剂量的许多分析是本领域已知的。出于本发明的目的，一种包括以下的分析可用于确定向哺乳动物施用的起始剂量:在一组哺乳动物中，每一只给予不同剂量的t细胞，比较当将给定剂量的所述t细胞给予哺乳动物时，靶细胞裂解的程度，或者表达本发明tcr、多肽或蛋白质的t细胞分泌ifn-γ的程度。可藉由本领域已知方法来分析施用特定剂量时靶细胞裂解的程度或所分泌的ifn-γ的程度。本发明tcr材料的剂量还通过施用特定的本发明tcr材料时可能伴随的任何不良副作用的存在、性质及程度来确定。通常，主治医生将决定用于治疗每例个体患者的本发明tcr材料的剂量，考虑各种因素，例如年龄、体重、一般健康状况、饮食、性别、拟给予的本发明tcr材料、给药途径和所治疗癌症的严重程度。在其中本发明tcr材料是细胞群的一个实施方案中，每次输注施用的细胞数量可变化，例如约1x106到约1x1012个细胞或更多。在某些实施例中，可施用少于1×106个细胞。本领域普通技术人员将容易了解，本发明的发明tcr材料可以任何多种方式修饰，以便通过修饰来提高本发明tcr材料的治疗或预防效果。例如，本发明tcr材料可直接或间接通过桥接与化学治疗剂偶联。将化合物偶联到化学治疗剂的实践在本领域是已知的。本领域普通技术人员认识到，对于本发明tcr材料的功能而言不必需的本发明tcr材料上的部位是连接桥接和/或化学治疗剂的理想部位，前提是桥接和/或化学治疗剂一旦连接到本发明tcr材料上，就不会干扰本发明tcr材料的功能，即与突变型p53结合或检测、治疗或预防癌症的能力。预期本发明的药物组合物、tcr、多肽、蛋白质、核酸、重组表达载体、宿主细胞或细胞群可用于治疗或预防癌症的方法。在不约束于特定理论的情况下，认为本发明的tcr特异性结合于突变的p53，使得当由细胞表达时，tcr(或相关的本发明的多肽或蛋白质)能够介导对表达突变p53的靶细胞的免疫应答。在这方面，本发明的实施方案提供治疗或预防哺乳动物中的癌症的方法，其包含以有效治疗或预防哺乳动物中癌症的量向哺乳动物施用本文所述的任一种药物组合物、tcr、多肽或蛋白质、包含编码本文所述的任一tcr、多肽或蛋白质的核苷酸序列的任何核酸或重组表达载体、或者任何宿主细胞或细胞群，其包含编码本文所述的任一tcr、多肽或蛋白质的重组载体。本发明的实施方案提供本文所述的任何药物组合物、tcr、多肽或蛋白质、包含编码本文所述的任何tcr、多肽、蛋白质的核苷酸序列的任何核酸或重组表达载体、或包含编码本文所述的任何tcr、多肽或蛋白质的重组载体的任何宿主细胞或细胞群，其用于治疗或预防哺乳动物中的癌症。如本文所使用的术语“治疗”和“预防”以及由此产生的词语不一定意味着100％或完全治疗或预防。而是存在不同程度的治疗或预防，本领域技术人员认识到其具有潜在的益处或治疗效果。在这方面，本发明方法可在哺乳动物中提供任何水平的癌症治疗或预防。此外，本发明方法提供的治疗或预防可包括对正在治疗或预防的癌症的一种或多种病况或症状的治疗或预防。例如，治疗或预防可包括促进肿瘤消退。此外，出于本文之目的，“预防”可包括延迟癌症或其症状或状况的发作。可选地或者另外地，“预防”可包括预防或延迟癌症或其症状或状况的复发。本发明实施方案还提供了检测哺乳动物中是否存在癌症的方法。该方法包括(i)使包含来自哺乳动物的一或多个细胞的样品与本文所述的任何本发明tcr、多肽、蛋白质、核酸、重组表达载体、宿主细胞、细胞群或药物组合物接触，由此形成复合物，并检测所述复合物，其中所述复合物的检测指示所述哺乳动物中存在癌症。关于在哺乳动物中检测癌症的本发明方法，细胞样本可以是包含全细胞、其裂解物或全细胞裂解物的部分，例如，核或细胞质部分、全蛋白部分或核酸部分的样品。出于本发明检测方法的目的，对于哺乳动物，可在体外或体内进行接触。优选地，该接触是在体外。此外，可通过本领域已知的任何多种方式来检测该复合物。例如，本文所述的本发明的tcr、多肽、蛋白质、核酸、重组表达载体、宿主细胞或细胞群，可以用可检测的标签标记，例如，放射性同位素、荧光团(例如，异硫氰酸荧光素(fitc)、藻红蛋白(pe))，酶(例如碱性磷酸酶、辣根过氧化物酶)和元素颗粒(例如金颗粒)。出于本发明方法的目的，其中施用宿主细胞或细胞群，所述细胞可以是哺乳动物同种异体或自体的。优选地，细胞是哺乳动物自体的。就本发明方法而言，癌症可为任何癌症，包括急性淋巴细胞癌、急性髓性白血病、肺泡横纹肌肉瘤、骨癌、脑癌、乳腺癌、肛门癌、肛管或肛门直肠癌、眼癌、肝内胆管癌、关节癌、颈部癌、胆囊或胸膜癌、鼻、鼻腔或中耳癌、口腔癌、阴道癌、外阴癌、慢性淋巴细胞性白血病、慢性髓系癌、结肠癌、结直肠癌、子宫内膜癌、食管癌、子宫颈癌、胃肠道类癌、胶质瘤、霍奇金淋巴瘤、下咽癌、肾癌、喉癌、肝癌、肺癌、恶性间皮瘤、黑色素瘤、多发性骨髓瘤、鼻咽癌、非霍奇金淋巴瘤、口咽癌、卵巢癌、阴茎癌、胰腺癌、腹膜、网膜和肠系膜癌、咽癌、前列腺癌、直肠癌、肾癌、皮肤癌、小肠癌、软组织癌、胃癌、睾丸癌、甲状腺癌、子宫癌、输尿管癌和膀胱癌。在一个优选的实施方案中，该癌症是表达突变p53的癌症。癌症可在seqidno:1的位置175、220、245、248、249、273和282中的任何一个或多个处表达具有突变的p53。癌症可表达具有以下一种或多种人类p53突变的p53:r175h、y220c、g245d、g245s、r248l、r248q、r248w、r249s、r273h、r273c、r273l和r282w。优选地，癌症是上皮癌或胆管癌、黑色素瘤、结肠癌、直肠癌、卵巢癌、子宫内膜癌、非小细胞肺癌(nsclc)、胶质母细胞瘤、子宫颈癌、头颈癌、乳腺癌、胰腺癌或膀胱癌。本发明方法中提及的哺乳动物可以是任何哺乳动物。如本文所使用，术语“哺乳动物”是指任何哺乳动物，包括但不限于啮齿目的哺乳动物，例如小鼠和仓鼠，和兔形目的哺乳动物，例如兔。优选哺乳动物来自食肉目，包括猫(猫)和犬(犬)。更优选的是，哺乳动物来自偶蹄目，包括bovines(奶牛)和swines(猪)；或来自persodactyla目，包括equines(马)。最优选地，哺乳动物属于灵长动物目，ceboids，或simoids(猴子)或类人猿目(人类和类人猿)。特别优选的哺乳动物是人类。以下实例进一步说明本发明，但当然不应以任何方式解释为限制其范围。实施例在以下实施例中所述的实验中使用表1-3中列出的氨基酸序列。在表1-2中，“lp”代表“长肽”。在表3中，“tmg”代表“串联微基因”。表1表2列出了表1的肽的野生型形式。表2表3实施例1该实施例证实了来自患者4127的4个抗突变p53的tcr的分离和特异的反应性。结合图1-7、36和37a-37c所述对患者4127进行实验。通过美国专利申请号2017/0224800(“tran方法”)中描述的方法鉴别该患者的突变p53反应性t细胞。分离单个tcr的方法如下所述。使用图1中所示的其中一种tcr转导自体pbl，并在2周后进行检测。将自体dc细胞以3x104个细胞/孔铺板，并用逐渐降低浓度的wtp53-g245肽(wt)或突变型p53-g245s肽(mut)脉冲过夜。每孔添加3x104个转导的t细胞并且在37℃下共培养过夜。收获上清液，用于ifn-γelisa。ifn-γelisa结果见图1。通过facs设门测定4-1bb表达:淋巴细胞\pi(neg)cd3+\cd3+mtcr+\cd8(neg)cd4+。facs结果见图2。用图3中所示的一种tcr转导自体pbl，并按照图2中所述的实验进行共培养，但dcs用dmso(肽媒介物)或表a中所示的具有重叠的14个氨基酸的15-merp53-g245s肽之一脉冲。如图2所述进行4-1bb表达。结果见图3。表a将cos7细胞(每孔2.5x104个)涂铺在平底96孔板的孔上。20小时后，将细胞用单独的hla等位基因共转染。20小时后，将细胞用10μg/ml的p53g245s-15-mer肽在37℃下脉冲3小时。洗涤后，将t细胞(105)加入孔中，并于37℃共培养过夜(20小时)。通过elisa法检测ifn-γ的分泌。通过netmhciipan预测:cbs.dtu.dk/services/netmhciipan/。通过facs测量4-1bb表达。facs设门:淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)→cd4+(4127-til)或cd4+mtcr+(tcr转导的t细胞)。结果显示在图4-5和表b中。表b在ncihla数据库中，3719例drb_型患者中有1367例(37％)表达drb3*02，在nci-sb(美国国家癌症研究所外科分会)的子宫内膜癌和卵巢癌患者中有5/9(56％)表达drb3*02。根据allelefrequencies.netwebsite，报告的drb3*02等位基因频率非常高。例如，本网站报告drb3*02等位基因的频率是美国nmdp中东或非洲北海岸人口的0.3447。来自患者4127的til与同种异体(drb3*01:01:01或drb3*02:02:01)抗原呈递细胞(apc)共培养，所述抗原呈递细胞(1)用由无关、wtp53或突变的p53序列组成的串联微型基因(tmg)电穿孔，或者(2)用肽媒介物(dmso)或由wtp53-g245s序列或突变的p53-g245s序列组成的经纯化的(>95％，通过高效液相色谱(hplc))25个氨基酸的肽脉冲。在37℃下进行共培养过夜。使用elispot测定评价ifn-γ的分泌。结果示于图6中。对p53-g245s特异的、表达4127-tcr1的t细胞与同种异体apc共培养，如图6实验所述。结果示于图7中。自体apc用浓度逐渐降低的对应于wtp53-g245或突变型p53-g245s序列的25个氨基酸的肽脉冲。用来自患者4127的4127-o37-tcr转导的t细胞与肽脉冲的apc在37℃共培养过夜。对于淋巴细胞→单细胞→活细胞)→cd3+mtcr+设门后，通过流式细胞术评价4-1bb的表达。结果见图36。将cos7细胞(2.5x104/孔)铺在平底96孔板的孔上。第二天，将细胞与来自患者4127的单个hla等位基因共转染。第二天，在转染的cos7细胞上脉冲25个氨基酸的p53-g245s肽。洗去过量的肽。t细胞用来自患者4127的一种tcr转导。向与cos7细胞共培养物中加入转导的t细胞(2x104个细胞/孔)。共培养物在37℃下孵育过夜。采用elisa法评价ifn-γ的分泌。对于淋巴细胞→单细胞→活细胞)→cd3+mtcr+设门后，通过流式细胞术评价4-1bb的表达。结果见图37a-37c。从患者4127中分离的tcr4127-tp53-g245s-tcr1的序列如下所述。从氨基末端开始，第一个下划线区域为cdr1β(seqidno:30)，第二个下划线区域为cdr2β(seqidno:31)，第三个下划线区域为cdr3β(seqidno:32)，第四个下划线区域为cdr1α(seqidno:27)，第五个下划线区域为cdr2α(seqidno:28)，第六个下划线区域为cdr3α(seqidno:29)。粗体区域为连接子(seqidno:26)。从氨基末端开始，第一个斜体区域是β链恒定区(seqidno:25)，第二个斜体区域是α链恒定区(seqidno:23)。β链可变区(seqidno:34)包括从氨基末端开始并且就在β链恒定区开始之前结束的序列。α链可变区(seqidno:33)包括连接子后立即开始并且就在α链恒定区开始之前结束的序列。全长β链(seqidno:36)包括从氨基末端开始并且就连接子开始前结束的序列。全长α链(seqidno:35)包括在连接子之后立即开始并且以羧基末端结束的序列。如下所述鉴定癌症反应性t细胞。如下所述分离tcr。tcr名称:4127-tp53-g245s-tcr1p53突变的识别:g245s方法:tran法共培养以鉴定tcr:与g245s长肽共培养4127-f10til片段，分选cd4+41bb+til细胞(仅单细胞rt-pcr)鉴定tcr的方法:单细胞rt-pcr(见下文)，适应性pairseq(4127-f10片段)，来自til克隆o71的5'race所有配对tcr中tcr的丰度:31.3％(在16对中观察到5次)tcr方向:β-α表达载体:γ-逆转录病毒患者4127的tcr4127-tp53-g245s-tcr1的统计数据如下表4所示。表4参数#频率总孔数96100％cdr3α型66.3％cdr3β1313.5％4127-tp3-g245s-tcr1对55.2％总配对的tcr1616.7％从患者4127中分离的tcr4127-tp53-g245s-tcr4的序列如下所述。从氨基末端开始，第一个下划线区域为cdr1β(seqidno:40)，第二个下划线区域为cdr2β(seqidno:41)，第三个下划线区域为cdr3β(seqidno:42)，第四个下划线区域为cdr1α(seqidno:37)，第五个下划线区域为cdr2α(seqidno:38)，第六个下划线区域为cdr3α(seqidno:39)。粗体区域为连接子(seqidno:26)。从氨基末端开始，第一个斜体区域是β链恒定区(seqidno:25)，第二个斜体区域是α链恒定区(seqidno:23)。β链可变区(seqidno:44)包括从氨基末端开始并且就在β链恒定区开始前结束的序列。α链可变区(seqidno:43)包括在连接子后立即开始并且就在α链恒定区开始前结束的序列。全长β链(seqidno:46)包括从氨基末端开始并且就在连接子开始前结束的序列。全长α链(seqidno:45)包括在连接子之后立即开始并且以羧基末端结束的序列。如下所述鉴定癌症反应性t细胞。如下所述分离tcr。tcr名称:4127-tp53-g245s-tcr4p53突变的识别:g245s方法:tran法共培养以鉴定tcr:与g245s长肽共培养4127-f10til片段，分选cd4+41bb+til细胞(仅单细胞rt-pcr)鉴定tcr的方法:单细胞rt-pcr(见下文)，适应性pairseq(4127-f10片段)所有配对tcr中tcr的丰度:12.5％(在16对中观察到2次)tcr方向:β-α表达载体:γ-逆转录病毒患者4127的tcr4127-tp53-g245s-tcr4统计数据见下表5。表5参数#频率总孔数96100％cdr3α基因22.1％cdr3β基因3940.6％4127-tp53-g245s-tcr4对22.1％总配对的tcr1616.7％从患者4127中分离的tcr4127_o102_tcr的序列如下所述。从氨基末端开始，第一个下划线区域为cdr1β(seqidno:50)，第二个下划线区域为cdr2β(seqidno:51)，第三个下划线区域为cdr3β(seqidno:52)，第四个下划线区域为cdr1α(seqidno:47)，第五个下划线区域为cdr2α(seqidno:48)，第六个下划线区域为cdr3α(seqidno:49)。粗体区域为连接子(seqidno:26)。从氨基末端开始，第一个斜体区域是β链恒定区(seqidno:25)，第二个斜体区域是α链恒定区(seqidno:23)。β链可变区(seqidno:54)包括从氨基末端开始并在β链恒定区开始前结束的序列。α链可变区(seqidno:53)包括连接子后立即开始并且就在α链恒定区开始前即结束的序列。全长β链(seqidno:56)包括从氨基末端开始并且在连接子开始前即结束的序列。全长α链(seqidno:55)包括在连接子之后立即开始并且以羧基末端结束的序列。如下所述确定癌症反应性t细胞。如下所述，分离tcr。tcr名称:4127_o102_tcrp53突变的识别:g245s方法:tran法共培养以鉴定tcr:未进行。使用5'race由t细胞克隆(o102)的rna直接鉴定tcr。通过分选ox40+t细胞以及通过有限稀释确立t细胞克隆来发展克隆，使用tran方法筛选t细胞克隆。鉴定tcr的方法:5’race，从til克隆o102所有配对的tcr中的tcr的丰度:100％t细胞克隆(不适用)tcr取向:α-β表达载体:γ-逆转录病毒从患者4127中分离的tcr4127-o37-tcr的序列如下所述。从氨基末端开始，第一个带下划线的区域是cdr1β(seqidno:460)，第二个带下划线的区域是cdr2β(seqidno:461)，第三个带下划线的区域是cdr3β(seqidno:462)，第四个带下划线的区域是cdr1α(seqidno:457)，第五个下划线区域为cdr2α(seqidno:458)，第六个下划线区域为cdr3α(seqidno:459)。粗体区域为连接子(seqidno:26)。从氨基末端开始，第一个斜体区域是β链恒定区(seqidno:25)，第二个斜体区域是α链恒定区(seqidno:23)。β链可变区(seqidno:464)包括从氨基末端开始并在β链恒定区开始前结束的序列。α链可变区(seqidno:463)包括连接子后立即开始并在α链恒定区开始前即结束的序列。全长β链(seqidno:466)包括从氨基末端开始到连接子开始前即结束的序列。全长α链(seqidno:465)包括在连接子之后立即开始并且以羧基末端结束的序列。如下所述确定癌症反应性t细胞。如下所述，分离tcr。tcr名称:4127-o37-tcr识别p53突变的识别:g245s方法:tran法共培养以鉴别tcr:未进行。使用适应性tcrad和tcrb测量法(下一代测序(ngs)平台)从t细胞克隆(o37)的基因组dna中直接鉴别tcr。通过分选ox40+t细胞以及通过有限稀释确立t细胞克隆来发展o37克隆。使用tran方法筛选t细胞克隆。鉴别tcr的方法:til克隆o37的适应性tcr测序(ngs)所有配对tcr中的tcr丰度:100％t细胞克隆(n/a)tcr方向:β-α表达载体:γ-逆转录病毒实施例2该实施例证实了来自患者4196的3种抗突变p53tcr的分离及特异性反应性。按照图8-11所述，对患者4196进行实验。如美国专利申请号2017/0224800所述，通过tran方法鉴定该患者的p53反应性t细胞。使用fluidigm方法分离单独tcr。患者4196的统计数据见表d。p53反应性细胞的特征:鉴定最小表位的挑战:第一个具有突变氨基酸p53r175h的预测肽如表c所示。表chlahla-a0201肽hmtevvrhc(seqidno:530)核心hmtevvrhc(seqidno:530)亲和力(nm)7826.06％等级13.00mg1vs4196-rx1til:cos7细胞用质粒转染tmg1和hla等位基因。细胞与4196rx1til输液袋共培养20小时(h)。结果见图8a。使用候选最小表位(hmtevvrhc(seqidno:530)或(sqhmtevrh(seqidno:531))脉冲p53-r175h最小肽versus4196-rx1til:第4天自体dcs2小时。清洗细胞并用4196rx1til输液袋共培养20小时。结果见图8b。a*02:01限制性tp53四聚体可用于从til输液袋中分离tp53反应性细胞:与tp53肽共同培养后使用4-1bb+分选分离tp53反应性细胞的技术有限。为分离tp53反应性细胞，生成a*02:01-tp53四聚体。生成不相关四聚体(a*02:01-gp100)作为对照。对活cd3+、cd8+细胞设门进行facs分析。对于a*02:01-tp53四聚体，检测到14.9％的p53四聚体+细胞。对于a*02:01-gp100四聚体，检测到0.7％的p53四聚体+细胞。tp53反应性细胞的分离和表征:tp53反应性细胞的分离和表征策略如下:(1)如上所述使用facs对四聚体阳性细胞进行分选。(2)进行单细胞pcr。(3)进行vβ渗出序列测定。使用单细胞pcr鉴定了tp53tcr的几个候选克隆，如表d所示。tcr-1a和tcr-1b是可选候选克隆。表d用msgv1载体克隆编码候选tcr的质粒。使用逆转录病毒转导将tcr引入供体淋巴细胞中。第4天，在a*02:01dc上，对hplc级最小表位脉冲2小时。dc细胞与tcr转导的细胞共培养20h。结果见图9a-9d。鉴定了三种不同的a*02:01限制性tp53受体。将t2细胞解冻并静置24小时，然后用浓度逐渐降低的hplc最小表位脉冲2小时。然后洗涤细胞并与tp53tcr转导的细胞共培养17小时。结果见图10a-10b。使用hla-a*02:01转导以下亲代肿瘤细胞系:结肠系–ls123(atccccl-255)，有r175h，但hla-a2状态未知；白血病细胞系:ccrf-cem(atcccrm-ccl-119，白血病)，有r175h，但hla-a2状态未知；乳腺系:au-565(atcccrl-2351，乳腺腺癌)，有r175h，但hla-a2状态未知；黑色素瘤细胞系:mel624，内源性表达hla-a2和wtr175。在共同培养的早上收获靶向转导的肿瘤细胞并铺板(1x105个细胞/孔)。在靶细胞上，脉冲r175h最小表位(hplc，5μg/ml)2h。洗涤细胞两次，并与2x104tp53tcr转导的细胞共培养20小时(患者4196)。使用ifn-γelispot读取应答。结果见图11。关于本实施例的靶细胞hla-a2和tp53表达和tcr反应性的其他实验数据在实施例15中提供。从患者4196中分离的tcr4196_av12-1_with_bv6-1如下所述。从氨基末端开始，第一个下划线区域为cdr1β(seqidno:70)，第二个下划线区域为cdr2β(seqidno:71)，第三个下划线区域为cdr3β(seqidno:72)，第四个下划线区域为cdr1α(seqidno:67)，第五个下划线区域为cdr2α(seqidno:68)，第六个下划线区域为cdr3α(seqidno:69)。粗体区域为连接子(seqidno:26)。从氨基末端开始，第一个斜体区域是β链恒定区(seqidno:25)，第二个斜体区域是α链恒定区(seqidno:24)。β链可变区(seqidno:74)包括从氨基末端开始并在β链恒定区开始前即结束的序列。α链可变区(seqidno:73)包括连接子后立即开始并在α链恒定区开始前即结束的序列。全长β链(seqidno:76)包括从氨基末端开始到连接子开始前即结束的序列。全长α链(seqidno:75)包括在连接子之后立即开始并且以羧基末端结束的序列。如下所述鉴定癌症反应性t细胞。tcr名称:4196_av12-1_with_bv6-1p53突变的识别:r175h方法:tran法从患者4196中分离的tcr4196_av38-1_with_bv10-3如下所述。从氨基末端开始，第一个下划线区域为cdr1β(seqidno:80)，第二个下划线区域为cdr2β(seqidno:81)，第三个下划线区域为cdr3β(seqidno:82)，第四个下划线区域为cdr1α(seqidno:77)，第五个下划线区域为cdr2α(seqidno:78)，第六个下划线区域为cdr3α(seqidno:79)。粗体区域为连接子(seqidno:26)。从氨基末端开始，第一个斜体区域是β链恒定区(seqidno:25)，第二个斜体区域是α链恒定区(seqidno:24)。β链可变区(seqidno:84)包括从氨基末端开始并在β链恒定区开始即前结束的序列。α链可变区(seqidno:83)包括连接子后立即开始和α链恒定区开始前即结束的序列。全长β链(seqidno:86)包括从氨基末端开始到连接子开始前即结束的序列。全长α链(seqidno:85)包括在连接子之后立即开始并且以羧基末端结束的序列。如下所述鉴定癌症反应性t细胞。tcr名称:4196_av38-1_with_bv10-3p53突变的识别:r175h方法:tran法从患者4196中分离的tcr4196_av6_with_bv11-2如下所述。从氨基末端开始，第一个下划线区域为cdr1β(seqidno:90)，第二个下划线区域为cdr2β(seqidno:91)，第三个下划线区域为cdr3β(seqidno:92)，第四个下划线区域为cdr1α(seqidno:87)，第五个下划线区域为cdr2α(seqidno:88)，第六个下划线区域为cdr3α(seqidno:89)。粗体区域为连接子(seqidno:26)。从氨基末端开始，第一个斜体区域是β链恒定区(seqidno:25)，第二个斜体区域是α链恒定区(seqidno:24)。β链可变区(seqidno:94)包括从氨基末端开始并在β链恒定区开始前即结束的序列。α链可变区(seqidno:93)包括连接子后立即开始且α链恒定区开始前即结束的序列。全长β链(seqidno:96)包括从氨基末端开始到连接子开始前即结束的序列。全长α链(seqidno:95)包括在连接子之后立即开始并且以羧基末端结束的序列。如下所述鉴定癌症反应性t细胞。tcr名称:4196_av6_with_bv11-2识别p53突变:r175h方法:tran法实施例3该实例证实了来自患者4238的11个抗突变p53tcr的分离。如对于图12-15所述对患者4238进行实验。患者4238的til片段(f1-f24,n＝24)与由不相关或突变的p53序列组成的tmg电穿孔的自体apc共培养。共培养物在37℃下培养过夜。使用elispot试验评估ifn-γ的分泌。结果见图12。对淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)设门后，通过流式细胞术评价4-1bb的表达。结果见图14。患者4238的til片段(f1-f24，n＝24)与自体apc共培养，所述自体apc用肽媒介物(dmso)脉冲或用由突变型p53-r248q序列组成的纯化的25个氨基酸的肽(hplc法>95％)脉冲。共培养物在37℃下培养过夜。使用elispot试验评估ifn-γ的分泌。结果见图13。对淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)设门后，通过流式细胞术评价4-1bb的表达。结果见图15。til片段f10、f11和f17是cd8+41bb+f17分选后tcr的来源。从患者4238中分离的tcr4238-f10-tcr1的序列如下所述。从氨基末端开始，第一个下划线区域为cdr1α(seqidno:207)，第二个下划线区域为cdr2α(seqidno:208)，第三个下划线区域为cdr3α(seqidno:209)，第四个下划线区域为cdr1β(seqidno:210)，第五个下划线区域为cdr2β(seqidno:211)，第六个下划线区域为cdr3β(seqidno:212)。粗体区域为连接子(seqidno:26)。从氨基末端开始，第一个斜体区域是α链恒定区(seqidno:23)，第二个斜体区域是β链恒定区(seqidno:25)。α链可变区(seqidno:213)包括从氨基末端开始并在α链恒定区开始前结束的序列。β链可变区(seqidno:214)包括连接子后立即开始并在β链恒定区开始前即结束的序列。全长α链(seqidno:215)包括从氨基末端开始到连接子开始前即结束的序列。全长β链(seqidno:216)包括在连接子之后立即开始并且以羧基末端结束的序列。使用下述筛选法鉴定癌症反应性t细胞。用于分离tcr的方法如下所述。tcr名称:4238-f10-tcr1p53突变的识别:r248q筛选法:p53“热点”突变通用筛选共培养以鉴定tcr:与r248q长肽共培养4238-f10，分选cd8+41bb+nmt鉴定tcr的方法:单细胞逆转录酶聚合酶链反应(rt-pcr)所有配对的tcr中的tcr的丰度:25.0％(在12对中观察到3次)tcr取向:α-β表达载体:sb转座子患者4238的tcr4238-f10-tcr1统计见下表6。表6参数#频率总孔数96100％cdr3α44.2％cdr3β44.2％4238-f10-tcr1对33.1％总配对tcr1212.5％从患者4238中分离的tcr4238-f10-tcr2的序列如下所述。从氨基末端开始，第一个下划线区域为cdr1α(seqidno:217)，第二个下划线区域为cdr2α(seqidno:218)，第三个下划线区域为cdr3α(seqidno:219)，第四个下划线区域为cdr1β(seqidno:220)，第五个下划线区域为cdr2β(seqidno:221)，第六个下划线区域为cdr3β(seqidno:222)。粗体区域为连接子(seqidno:26)。从氨基末端开始，第一个斜体区域是α链恒定区(seqidno:23)，第二个斜体区域是β链恒定区(seqidno:25)。α链可变区(seqidno:223)包括从氨基末端开始并在α链恒定区开始前即结束的序列。β链可变区(seqidno:224)包括在连接子之后立即开始并在β链恒定区开始之前即结束的序列。全长α链(seqidno:225)包括从氨基末端开始到连接子开始前即结束的序列。全长β链(seqidno:226)包括在连接子之后立即开始并且以羧基末端结束的序列。使用下述筛选方法鉴定癌症反应性t细胞。用于分离tcr的方法如下所述。tcr名称:4238-f10-tcr2识别p53突变的识别:r248q筛选方法:p53“热点”突变通用筛选共培养以鉴定tcr:与r248q长肽共培养4238-f10，分选cd8+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:25.0％(在12对中观察到3次)tcr取向:α-β表达载体:sb转座子患者4238的tcr4238-f10-tcr2统计见下表7。表7参数#频率总孔数96100％cdr3α44.2％cdr3β44.2％4238-f10-tcr2对33.1％总配对tcr1212.5％从患者4238中分离的tcr4238-f10-tcr3的序列如下所述。从氨基末端开始，第一个下划线区域为cdr1α(seqidno:227)，第二个下划线区域为cdr2α(seqidno:228)，第三个下划线区域为cdr3α(seqidno:229)，第四个下划线区域为cdr1β(seqidno:230)，第五个下划线区域为cdr2β(seqidno:231)，第六个下划线区域为cdr3β(seqidno:232)。粗体区域为连接子(seqidno:26)。从氨基末端开始，第一个斜体区域是α链恒定区(seqidno:23)，第二个斜体区域是β链恒定区(seqidno:25)。α链可变区(seqidno:233)包括从氨基末端开始并在α链恒定区开始前结束的序列。β链可变区(seqidno:234)包括在连接子后立即开始并在β链恒定区开始前即结束的序列。全长α链(seqidno:235)包括从氨基末端开始并在连接子开始前即结束的序列。全长β链(seqidno:236)包括在连接子后立即开始并以羧基末端结束的序列。使用下述筛选方法鉴别癌症反应性t细胞。用于分离tcr的方法如下所述。tcr名称:4238-f10-tcr3p53突变的识别:r248q筛选法:p53“热点”突变通用筛选共培养以鉴定tcr:与r248q长肽共培养4238-f10，分选cd8+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:58.3.0％(在12对中观察到7次)tcr取向:α-β表达载体:sb转座子患者4238的tcr4238-f10-tcr3统计见下表8。表8参数#频率总孔数96100％cdr3α1616.7％cdr3β2627.1％4238-f10-tcr3对77.3％总配对tcr1212.5％从患者4238中分离的tcr4238-f10-tcr4的序列如下所述。从氨基末端开始，第一个下划线区域为cdr1α(seqidno:237)，第二个下划线区域为cdr2α(seqidno:238)，第三个下划线区域为cdr3α(seqidno:239)，第四个下划线区域为cdr1β(seqidno:240)，第五个下划线区域为cdr2β(seqidno:241)，第六个下划线区域为cdr3β(seqidno:242)。粗体区域为连接子(seqidno:26)。从氨基末端开始，第一个斜体区域是α链恒定区(seqidno:23)，第二个斜体区域是β链恒定区(seqidno:25)。α链可变区(seqidno:243)包括从氨基末端开始到α链恒定区开始之前即结束的序列。β链可变区(seqidno:244)包括在连接子之后立即开始并在β链恒定区开始之前即结束的序列。全长α链(seqidno:245)包括从氨基末端开始到连接子开始前即结束的序列。全长β链(seqidno:246)包括在连接子后立即开始并以羧基末端结束的序列。使用下述筛选方法鉴定癌症反应性t细胞。用于分离tcr的方法如下所述。tcr名称:4238-f10-tcr4p53突变的识别:r248q筛选法:p53“热点”突变通用筛选共培养以鉴定tcr:与r248q长肽共培养4238-f10，分选cd8+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:58.3.0％(在12对中观察到7次)tcr取向:α-β表达载体:sb转座子患者4238的tcr4238-f10-tcr4统计见下表9。表9从患者4238中分离的tcr4238-f11-tcr1的序列如下所述。从氨基末端开始，第一个下划线区域为cdr1α(seqidno:247)，第二个下划线区域为cdr2α(seqidno:248)，第三个下划线区域为cdr3α(seqidno:249)，第四个下划线区域为cdr1β(seqidno:250)，第五个下划线区域为cdr2β(seqidno:251)，第六个下划线区域为cdr3β(seqidno:252)。粗体区域为连接子(seqidno:26)。从氨基末端开始，第一个斜体区域是α链恒定区(seqidno:23)，第二个斜体区域是β链恒定区(seqidno:25)。α链可变区(seqidno:253)包括从氨基末端开始并在α链恒定区开始前结束的序列。β链可变区(seqidno:254)包括连接子后立即开始并β链恒定区开始前即结束的序列。全长α链(seqidno:255)包括从氨基末端开始到连接子开始前即结束的序列。全长β链(seqidno:256)包括在连接子之后立即开始并且以羧基末端结束的序列。使用下述筛选方法鉴定癌症反应性t细胞。用于分离tcr的方法如下所述。tcr名称:4238-f11-tcr1p53突变的识别:r248q筛选法:p53“热点”突变通用筛选共培养以鉴定tcr:与r248q长肽共培养4238-f10，分选cd8+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:25.0％(在12对中观察到3次)tcr取向:α-β表达载体:sb转座子患者4238的tcr4238-f11-tcr1统计见下表10。表10参数#频率总孔数96100％cdr3α55.2％cdr3β33.1％4238-f11-tcr1对33.1％总配对tcr1212.5％从患者4238中分离的tcr4238-f11-tcr2的序列如下所述。从氨基末端开始，第一个下划线区域为cdr1α(seqidno:257)，第二个下划线区域为cdr2α(seqidno:258)，第三个下划线区域为cdr3α(seqidno:259)，第四个下划线区域为cdr1β(seqidno:260)，第五个下划线区域为cdr2β(seqidno:261)，第六个下划线区域为cdr3β(seqidno:262)。粗体区域为连接子(seqidno:262)。从氨基末端开始，第一个斜体区域是α链恒定区(seqidno:23)，第二个斜体区域是β链恒定区(seqidno:25)。α链可变区(seqidno:263)包括从氨基末端开始并在α链恒定区开始前结束的序列。β链可变区(seqidno:264)包括连接子后立即开始并在β链恒定区开始前即结束的序列。全长α链(seqidno:265)包括从氨基末端开始到连接子开始前即结束的序列。全长β链(seqidno:266)包括在连接子之后立即开始并且以羧基末端结束的序列。使用下述筛选方法鉴别癌症反应性t细胞。用于分离tcr的方法如下所述。tcr名称:4238-f11-tcr2p53突变的识别:r248q筛选法:p53“热点”突变通用筛选共培养以鉴定tcr:与r248q长肽共培养4238-f11，分选cd8+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:25.0％(在12对中观察到3次)tcr取向:α-β表达载体:sb转座子患者4238的tcr4238-f11-tcr2统计见下表11。表11参数#频率总孔数96100％cdr3α55.2％cdr3β88.3％4238-f11-tcr2对33.1％总配对tcr1212.5％从患者4238中分离的tcr4238-f11-tcr3的序列如下所述。从氨基末端开始，第一个下划线区域为cdr1α(seqidno:267)，第二个下划线区域为cdr2α(seqidno:268)，第三个下划线区域为cdr3α(seqidno:269)，第四个下划线区域为cdr1β(seqidno:270)，第五个下划线区域为cdr2β(seqidno:271)，第六个下划线区域为cdr3β(seqidno:272)。粗体区域为连接子(seqidno:26)。从氨基末端开始，第一个斜体区域是α链恒定区(seqidno:23)，第二个斜体区域是β链恒定区(seqidno:25)。α链可变区(seqidno:273)包括从氨基末端开始并在α链恒定区开始前即结束的序列。β链可变区(seqidno:274)包括连接子后立即开始并在β链恒定区开始前即结束的序列。全长α链(seqidno:275)包括从氨基末端开始到连接子开始前即结束的序列。全长β链(seqidno:276)包括在连接子之后立即开始并且以羧基末端结束的序列。使用下述筛选方法鉴别癌症反应性t细胞。用于分离tcr的方法如下所述。tcr名称:4238-f11-tcr3p53突变的识别:r248q筛选法:p53“热点”突变通用筛选共培养以鉴定tcr:与r248q长肽共培养4238-f11，分选cd8+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:16.7％(在12对中观察到2次)tcr取向:α-β表达载体:sb转座子患者4238的tcr4238-f11-tcr3统计见下表12。表12参数#频率总孔数96100％cdr3α44.2％cdr3β22.1％4238-f11-tcr3对22.1％总配对tcr1212.5％从患者4238中分离的tcr4238-f11-tcr4的序列如下所述。从氨基末端开始，第一个下划线区域为cdr1α(seqidno:277)，第二个下划线区域为cdr2α(seqidno:278)，第三个下划线区域为cdr3α(seqidno:279)，第四个下划线区域为cdr1β(seqidno:280)，第五个下划线区域为cdr2β(seqidno:281)，第六个下划线区域为cdr3β(seqidno:282)。粗体区域为连接子(seqidno:26)。从氨基末端开始，第一个斜体区域是α链恒定区(seqidno:23)，第二个斜体区域是β链恒定区(seqidno:25)。α链可变区(seqidno:283)包括从氨基末端开始并在α链恒定区开始前即结束的序列。β链可变区(seqidno:284)包括连接子后立即开始并在β链恒定区开始前即结束的序列。全长α链(seqidno:285)包括从氨基末端开始到连接子开始前即结束的序列。全长β链(seqidno:286)包括在连接子之后立即开始并以羧基末端结束的序列。使用下述筛选方法鉴别癌症反应性t细胞。用于分离tcr的方法如下所述。tcr名称:4238-f11-tcr4p53突变的识别:r248q筛选法:p53“热点”突变通用筛选共培养以鉴定tcr:与r248q长肽共培养4238-f11，分选cd8+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:16.7％(在12对中观察到2次)tcr取向:α-β表达载体:sb转座子患者4238的tcr4238-f11-tcr4统计见下表13。表13参数#频率总孔数96100％cdr3α44.2％cdr3β22.1％4238-f11-tcr4对22.1％总配对tcr1212.5％从患者4238中分离的tcr4238-f17-tcr1的序列如下所述。从氨基末端开始，第一个下划线区域为cdr1α(seqidno:287)，第二个下划线区域为cdr2α(seqidno:288)，第三个下划线区域为cdr3α(seqidno:289)，第四个下划线区域为cdr1β(seqidno:290)，第五个下划线区域为cdr2β(seqidno:291)，第六个下划线区域为cdr3β(seqidno:292)。粗体区域为连接子(seqidno:26)。从氨基末端开始，第一个斜体区域是α链恒定区(seqidno:23)，第二个斜体区域是β链恒定区(seqidno:25)。α链可变区(seqidno:293)包括从氨基末端开始并在α链恒定区开始前即结束的序列。β链可变区(seqidno:294)包括在连接子之后立即开始并在β链恒定区开始之前即结束的序列。全长α链(seqidno:295)包括从氨基末端开始到连接子开始前即结束的序列。全长β链(seqidno:296)包括在连接子之后立即开始并且以羧基末端结束的序列。使用下述筛选方法鉴别癌症反应性t细胞。用于分离tcr的方法如下所述。tcr名称:4238-f17-tcr1p53突变的识别:r248q筛选法:p53“热点”突变通用筛选共培养以鉴定tcr:与r248q长肽共培养4238-f11，分选cd8+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:33.3％(在6对中观察到2次)tcr取向:α-β表达载体:sb转座子患者4238的tcr4238-f17-tcr1统计见下表14。表14参数#频率总孔数96100％cdr3α33.1％cdr3β22.1％4238-f17-tcr1对22.1％总配对tcr66.3％从患者4238中分离的tcr4238-f17-tcr2的序列如下所述。从氨基末端开始，第一个下划线区域为cdr1α(seqidno:297)，第二个下划线区域为cdr2α(seqidno:298)，第三个下划线区域为cdr3α(seqidno:299)，第四个下划线区域为cdr1β(seqidno:300)，第五个下划线区域为cdr2β(seqidno:301)，第六个下划线区域为cdr3β(seqidno:302)。粗体区域为连接子(seqidno:26)。从氨基末端开始，第一个斜体区域是α链恒定区(seqidno:23)，第二个斜体区域是β链恒定区(seqidno:25)。α链可变区(seqidno:303)包括从氨基末端开始并在α链恒定区开始之前即结束的序列。β链可变区(seqidno:304)包括在连接子之后立即开始并在β链恒定区开始之前即结束的序列。全长α链(seqidno:305)包括从氨基末端开始到连接子开始前即结束的序列。全长β链(seqidno:306)包括在连接子之后立即开始并且以羧基末端结束的序列。使用下述筛选方法鉴别癌症反应性t细胞。用于分离tcr的方法如下所述。tcr名称:4238-f17-tcr2p53突变的识别:r248q筛选法:p53“热点”突变通用筛选共培养以鉴定tcr:与r248q长肽共培养4238-f11，分选cd8+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:33.3％(在6对中观察到2次)tcr取向:α-β表达载体:sb转座子患者4238的tcr4238-f17-tcr2统计见下表15。表15参数#频率总孔数96100％cdr3α66.3％cdr3β22.1％4238-f17-tcr2对22.1％总配对tcr66.3％从患者4238中分离的tcr4238-f17-tcr3的序列如下所述。从氨基末端开始，第一个下划线区域为cdr1α(seqidno:307)，第二个下划线区域为cdr2α(seqidno:308)，第三个下划线区域为cdr3α(seqidno:309)，第四个下划线区域为cdr1β(seqidno:310)，第五个下划线区域为cdr2β(seqidno:311)，第六个下划线区域为cdr3β(seqidno:312)。粗体区域为连接子(seqidno:26)。从氨基末端开始，第一个斜体区域是α链恒定区(seqidno:23)，第二个斜体区域是β链恒定区(seqidno:25)。α链可变区(seqidno:313)包括从氨基末端开始并在α链恒定区开始前即结束的序列。β链可变区(seqidno:314)包括在连接子之后立即开始并在β链恒定区开始之前即结束的序列。全长α链(seqidno:315)包括从氨基末端开始并在连接子开始之前即结束的序列。全长β链(seqidno:316)包括在连接子之后立即开始并且以羧基末端结束的序列。使用下述筛选方法鉴别癌症反应性t细胞。用于分离tcr的方法如下所述。tcr名称:4238-f17-tcr3p53突变的识别:r248q筛选法:p53“热点”突变通用筛选共培养以鉴定tcr:与r248q长肽共培养4238-f11，分选cd8+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:33.3％(在6对中观察到2次)tcr取向:α-β表达载体:sb转座子患者4238的tcr4238-f17-tcr3统计见下表16。表16实施例4该实施例证实了从患者4253中分离出两种抗突变p53tcr。按照图16所述，对患者4253进行实验。来自患者的til片段(f1-f24；n＝24)与自体apc共培养，所述自体apc(1)用肽媒介物(dmso)脉冲,(2)用由突变的p53-r248w序列组成的25个氨基酸的纯化肽(hplc法>95％)脉冲，(3)用不相关的tmg电穿孔或(4)用含有突变的p53-r248w序列的p53-mut-tmg电穿孔。共培养物在37℃下培养过夜。通过elispot评估ifn-γ的分泌。结果见图16。选择f15用于tcr的分选，因为它对p53-mut-tmg和p53-r248w长肽都有反应。从患者4253中分离的tcr4253-til-tcr1的序列如下所述。从氨基末端开始，第一个下划线区域为cdr1α(seqidno:187)，第二个下划线区域为cdr2α(seqidno:188)，第三个下划线区域为cdr3α(seqidno:189)，第四个下划线区域为cdr1β(seqidno:190)，第五个下划线区域为cdr2β(seqidno:191)，第六个下划线区域为cdr3β(seqidno:192)。粗体区域为连接子(seqidno:26)。从氨基末端开始，第一个斜体区域是α链恒定区(seqidno:23)，第二个斜体区域是β链恒定区(seqidno:25)。α链可变区(seqidno:193)包括从氨基末端开始并在α链恒定区开始前即结束的序列。β链可变区(seqidno:194)包括在连接子后立即开始并在β链恒定区开始前即结束的序列。全长α链(seqidno:195)包括从氨基末端开始到连接子开始前即结束的序列。全长β链(seqidno:196)包括在连接子之后立即开始并且以羧基末端结束的序列。使用下述筛选方法鉴别癌症反应性t细胞。用于分离tcr的方法如下所述。tcr名称:4253-til-tcr1p53突变的识别:r248w方法:简略的p53“热点”突变通用筛选(仅评价突变tmg和r248w长肽)共培养以鉴定tcr:与p53-r248w长肽共培养4253-f15，分选cd3+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:5.6％(36对中观察到2次)tcr取向:α-β表达载体:sb转座子患者4253的tcr4253-til-tcr1统计见下表17。表17参数#频率总孔数55100％cdr3α35.5％cdr3β23.6％4253-til-tcr1对23.6％总配对tcr3665.5％从患者4253中分离的tcr4253-til-tcr2的序列如下所述。从氨基末端开始，第一个下划线区域为cdr1α(seqidno:197)，第二个下划线区域为cdr2α(seqidno:198)，第三个下划线区域为cdr3α(seqidno:199)，第四个下划线区域为cdr1β(seqidno:200)，第五个下划线区域为cdr2β(seqidno:201)，第六个下划线区域为cdr3β(seqidno:202)。粗体区域为连接子(seqidno:26)。从氨基末端开始，第一个斜体区域是α链恒定区(seqidno:23)，第二个斜体区域是β链恒定区(seqidno:25)。α链可变区(seqidno:203)包括从氨基末端开始并在α链恒定区开始前即结束的序列。β链可变区(seqidno:204)包括在连接子后立即开始并在β链恒定区开始前即结束的序列。全长α链(seqidno:205)包括从氨基末端开始到连接子开始前即结束的序列。全长β链(seqidno:206)包括在连接子之后立即开始并且以羧基末端结束的序列。使用下述筛选方法鉴别癌症反应性t细胞。用于分离tcr的方法如下所述。tcr名称:4253-til-tcr2p53突变的识别:r248w方法:简略的p53“热点”突变通用筛选(仅评价突变tmg和r248w长肽)共培养以鉴定tcr:与p53-r248w长肽共培养4253-f15，分选cd3+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:91.7％(36对中观察到33次)tcr取向:α-β表达载体:sb转座子患者4253的tcr4253-til-tcr2统计见下表18。表18参数#频率总孔数55100％cdr3α3563.6％cdr3β3360.0％4253-til-tcr2对3360.0％总配对tcr3665.5％实施例5本实施例通过在自体t细胞内共培养经诱导以表达突变型p53的自体apc(“p53热点突变通用筛选”)来鉴定患者4273中的抗突变型p53t细胞。该实施例还证实从患者4273中分离出两种抗突变p53tcr。按照图17-20和56-60所述，对患者4273进行实验。将来自患者4273的til片段(f1-f24，n＝24)与用由不相关的、wtp53或突变型p53序列组成的tmg电穿孔的自体apc共培养。共培养物在37℃下培养过夜。使用elispot试验评估ifn-γ的分泌。结果见图17。对淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)设门后通过流式细胞术评价4-1bb的表达。结果见图18。患者4273的til片段(f1-f24,n＝24)与自体apc共培养，所述apc用肽媒介物(dmso)脉冲或者用由wtp53-r248序列或突变型p53-r248w序列组成的纯化(hplc法>95％)的25个氨基酸的肽脉冲。共培养物在37℃下过夜培养。使用elispot试验评估ifn-γ的分泌。结果见图19。对淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)设门后通过流式细胞术评价4-1bb的表达。结果见图20。对于患者4273，f15是最具反应性的片段，f15对lp和tmg的应答相当，主要是cd4t细胞。所以将4273-f15与用r248wlp脉冲的apc共培养，并在第二天将cd4+41bb+t细胞作为单细胞分选至96孔pcr板的孔中，每孔一个细胞。pcr板在每个孔中有rt-pcr溶液，可扩增相同溶液中的tcrα和βcdr3区域。然后将微孔板分成两个96孔pcr板，并进行第二次pcr反应以扩增cdr3α或cdr3β作为单独反应。通过sanger测序对各孔的pcr产物(共192个pcr产物映射至各孔-α或β)进行测序。将核苷酸序列输入imgt/v-quest(imgt.org/iview/vest)livret＝0&option＝humantcr),igblast(ncbi.nlm.nih.gov/igblast/igblast.cgi？cmd＝web和search_type＝tcr和link_loc＝igtab)，并由expasy翻译(web.exasy.org/transf/)。根据翻译序列确定变量家族并融合至cdr3和接合区(j或dj)。可变序列融合至鼠恒定序列，并且重构的tcrα和tcrβ通过弗林蛋白酶-柔性-p2a(rakr-sgsg-atnfsllkqagdveenpgp)(seqidno:26)连接。然后将该序列从头合成为dna，并克隆至表达载体(γ逆转录病毒或sleepingbeauty(sb)转座子(universityofminnesota,minneapolis,mn))中。然后使用标准病毒转导或非病毒转座方案使t细胞表达tcr，并针对推定的肽测试表达鼠源化tcr的t细胞(如通过小鼠tcrβ恒定链检测)。用编码无关突变、wtp53序列或经突变的p53序列(包括p53-r248w)的tmg转染自体apc。单独的培养基及pma和离子霉素分别为阴性对照和阳性对照。将来自患者4273的til(片段培养物8和15)与tmg转染的apc在37℃下共培养过夜。对于淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)设门后，通过流式细胞术评价4-1bb的表达。结果示于图56中。将自体apc在37℃下用对应于wt或经突变的p53-r248w新表位的25个氨基酸的肽脉冲2小时。将来自患者4273的对p53-r248w具有特异性的til(片段培养物15)与肽-脉冲的apc在37℃下共培养过夜。dmso是肽媒介物。对于淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)，设门后，通过流式细胞术评价4-1bb的表达。结果示于图57中。将自体apc用来自p53-r248w新表位的重叠14个氨基酸的15个氨基酸的肽脉冲。将来自患者4273的对p53-r248w具有特异性的til(片段培养物15)与肽-脉冲的apc在37℃下共培养过夜。dmso是肽媒介物，单独的培养基(仅t细胞)及pma和离子霉素是对照。25个氨基酸的肽(wtp53-r248和经突变的p53-r248w)是15个氨基酸的肽的另外对照。对于淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)→cd4+cd8-设门后，通过流式细胞术评价4-1bb的表达。结果示于图58中。将cos7细胞(每孔2.5x104个)涂铺在平底96孔板的孔上。第二天，将细胞分别与来自患者4273的单独的hla等位基因以及有或无p53-r248w新抗原的wt或经突变的tp53tmg共转染。第二天，将来自患者4273的对p53-r248w具有特异性的til(片段培养物15)与转染的cos7细胞在37℃下共培养过夜。通过elisa评价ifn-γ的分泌。结果示于图59中。将表达模拟物(无tcr)或4273-tcr1a2的t细胞与自体apc共培养，所述自体apc用肽媒介物(dmso)或由野生型p53-r248序列或经突变的p53-r248w序列组成的经纯化的(>95％，通过hplc)25个氨基酸的肽脉冲。单独的培养基及pma和离子霉素分别为阴性对照和阳性对照。在37℃下进行共培养过夜。通过elispot评价ifn-γ的分泌。结果示于图60中。从患者4273分离的tcr4273-tp53-r248w-tcr1a1的序列示于下文。从氨基末端开始，第一加下划线区是cdr1α(seqidno:437)，第二加下划线区是cdr2α(seqidno:438)，第三加下划线区是cdr3α(seqidno:439)，第四加下划线区是cdr1β(seqidno:440)，第五加下划线区是cdr2β(seqidno:441)，和第六加下划线区是cdr3β(seqidno:442)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一加斜体区是α链恒定区(seqidno:23)并且第二加斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:443)包含从氨基末端开始并且就在α链恒定区开始之前即结束的序列。β链可变区(seqidno:444)包含在连接子之后立即开始并且就在β链恒定区开始之前结束的序列。全长α链(seqidno:445)包含从氨基末端开始并且就在连接子开始之前结束的序列。全长β链(seqidno:446)包含在连接子之后立即开始并且以羧基末端结束的序列。tcr名称:4273-tp53-r248w-tcr1a1p53突变的识别:r248w方法:p53“热点”突变通用筛选从患者4273分离的tcr4273-tp53-r248w-tcr1a2的序列示于下文。从氨基末端开始，第一加下划线区是cdr1α(seqidno:447)，第二加下划线区是cdr2α(seqidno:448)，第三加下划线区是cdr3α(seqidno:449)，第四加下划线区是cdr1β(seqidno:450)，第五加下划线区是cdr2β(seqidno:451)，和第六加下划线区是cdr3β(seqidno:452)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一加斜体区是α链恒定区(seqidno:23)并且第二加斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:453)包含从氨基末端开始并且就在α链恒定区的开始之前结束的序列。β链可变区(seqidno:454)包含在连接子之后立即开始并且就在β链恒定区开始之前结束的序列。全长α链(seqidno:455)包含从氨基末端开始并且就在连接子开始之前结束的序列。全长β链(seqidno:456)包含在连接子之后立即开始并且以羧基末端结束的序列。tcr名称:4273-tp53-r248w-tcr1a2p53突变的识别:r248w方法:p53“热点”突变通用筛选下表19列出了患者4273的tcr的统计数据。在表19中，与经突变的p53蛋白(tmg或肽)共培养后，总有96个孔用41bb+t细胞分选。77个孔具有生产对(意指具有(1)序列和(2)该序列中没有终止密码子)，配对频率为80.2％。那些对中有43个是cdr3a/cdr3b组合，以构成4273-tp53-r248w-tcr1a1(生产对的55.8％)。那些对中有30个是cdr3a/cdr3b组合，以构成4273-tp53-r248w-tcr1a2(生产对的39％)。总体而言，在96个孔中，分别发现了4273-tp53-r248w-tcr1a1的cdr3a和cdr3b50次和83次。总体而言，在96个孔中，分别发现了4273-tp53-r248w-tcr1a2的cdr3a和cdr3b33次和83次。表19实施例6该实施例证实了在患者4149中抗-突变的p53t细胞的鉴定。该实施例还证实了来自患者4149的一种抗-突变的p53tcr的分离和特异反应性。如图21-24针对患者4149所述进行实验。使用tran方法发现了tcr。然后使用tcr来验证“p53热点突变通用筛选”方法。将tcr(4149-tcra2b1或4149-tcra2b2)转座至来自患者4149的自体pbl中，并与自体apc共培养，所述自体apc(1)用由无关、wtp53或经突变的p53序列组成的tmg电穿孔，或(2)用肽媒介物(dmso)或由wtp53-y220序列或经突变的p53-y220c序列组成的经纯化的(>95％，通过hplc)25-个氨基酸的肽脉冲。pma和离子霉素的组合是阳性对照。在37℃下进行共培养过夜。使用elispot测定评价ifn-γ的分泌。结果示于图21中。对于淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)→cd4+mtcr+(tcr转座的t细胞)设门后，通过流式细胞术评价4-1bb的表达。结果示于图22中。还进行了通过tcrad深度测序和tcrb深度测序确定cd4+4-1bb+细胞的百分比。结果示于表e中。表e被4149-f11识别的假定p53y220c最小表位的作图:将自体dc细胞经肽脉冲(10μg/ml)并在粒细胞-巨噬细胞集落刺激因子(gm-csf)和il-4中静置过夜。将til在500cu/mlil-2中静置2-3天。将2x104个til和105个靶细胞在37℃下共培养过夜。通过elispot测量ifn-γ。结果示于表10中。通过facs用设门淋巴细胞\pi(neg)cd3+\cd3+cd4+测量4-1bb表达。结果示于图23中。表f将cos7细胞(每孔2.5x104个)涂铺在平底96孔板的孔上。20小时后，将细胞与单独的hla等位基因在有或无tmg的情况下下共转染。20小时后，将自体dc细胞用tmg平行转染。将所有hlaii类等位基因在有或无tmg的情况下共转染一组孔中。将未用tmg转染的细胞在37℃下用10μg/ml的p53-y220c15-聚体肽脉冲2-3小时。洗涤后，将在第二rep的第14天的4149-tcra2b2-转座的t细胞(105)添加至孔，并在37℃下共培养过夜。通过elisa测量ifn-γ分泌。结果示于图24中。通过netmhciipan进行预测(表g):cbs.dtu.dk/services/netmhciipan/。表g在ncihla数据库中的3719名drb_型患者中有1367名(37％)以及在nci-sb的9名子宫内膜癌和卵巢癌患者中有5名(56％)检测到drb3*02表达。drb3*02等位基因的报道频率非常高，如实施例1所述。从患者4149分离的tcr4149tcra2b2的序列示于下文。从氨基末端开始，第一加下划线区是cdr1α(seqidno:57)，第二加下划线区是cdr2α(seqidno:58)，第三加下划线区是cdr3α(seqidno:59)，第四加下划线区是cdr1β(seqidno:60)，第五加下划线区是cdr2β(seqidno:61)，和第六加下划线区是cdr3β(seqidno:62)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一加斜体区是α链恒定区(seqidno:23)并且第二加斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:63)包含从氨基末端开始并且就在α链恒定区的开始之前结束的序列。β链可变区(seqidno:64)包含在连接子之后立即开始并且就在β链恒定区开始之前结束的序列。全长α链(seqidno:65)包含从氨基末端开始并且就在连接子开始之前结束的序列。全长β链(seqidno:66)包含在连接子之后立即开始并且以羧基末端结束的序列。癌症反应性t细胞使用下面列出的筛选方法进行鉴定。该患者的p53反应性细胞通过美国申请2017/0224800描述的transcreening方法来鉴定。tcr名称:4149tcra2b2p53突变的识别:y220c筛选方法:用于验证p53“热点”突变通用筛选共培养以鉴定tcr:共培养4149-f11til片段与p53-y220c长肽，分选cd4+41bb+t细胞鉴定tcr的方法:频率配对.将cd4+41bb+分选的t细胞在rep(快速扩增方案)中扩增并使所得的t细胞培养物经受adaptivebiotechnologies的tcrad(α)和tcrb(β)深度测序。在4种总tcr矩阵中，使前两种tcrα与前两种tcrβ配对。第二tcrα和第二tcrβ是反应性tcr，因此tcra2b2命名。所有tcr中tcr的丰度:如下tcr取向:α-β表达载体:sb转座子患者4149的tcr4149tcra2b2的统计数据示于下表20中。表20实施例7该实施例证实了通过在自体t细胞内共培养被诱导表达突变的p53的自体apc来鉴定患者4213中的抗-突变的p53t细胞(“p53热点突变通用筛选”)。该实施例还证实了从患者4213分离12种抗-突变的p53tcr。如图25-26针对患者4213所述进行实验。将来自患者4213的til片段(f2和f24)与用肽媒介物(dmso)或由经突变的p53-r248q序列组成的经纯化的(>95％，通过hplc)25-个氨基酸的肽脉冲的自体apc共培养。在37℃下进行共培养过夜。对于淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)设门后，通过流式细胞术评价4-1bb的表达。结果示于图25中。将cd8+4-1bb+t细胞分选至96孔板的孔中。使用单细胞rt-pcr鉴定tcr。cd4+t细胞来自患者4213的外周血淋巴细胞。将cd4+t细胞培养物与用肽媒介物(dmso)或由经突变的p53-r248q序列组成的经纯化的(>95％，通过hplc)25-个氨基酸的肽脉冲的自体apc共培养。在37℃下进行共培养过夜。ifn-γ的分泌通过elispot评价。对于淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)设门后，通过流式细胞术评价4-1bb的表达。结果示于图26中。将cd4+41bb+t细胞分选至96孔板的孔中。使用单细胞rt-pcr鉴定tcr。从患者4213分离的4213-f2-tcr1的序列示于下文。从氨基末端开始，第一加下划线区是cdr1α(seqidno:317)，第二加下划线区是cdr2α(seqidno:318)，第三加下划线区是cdr3α(seqidno:319)，第四加下划线区是cdr1β(seqidno:320)，第五加下划线区是cdr2β(seqidno:321)，和第六加下划线区是cdr3β(seqidno:322)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一加斜体区是α链恒定区(seqidno:23)并且第二加斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:323)包含从氨基末端开始并且就在α链恒定区的开始之前结束的序列。β链可变区(seqidno:324)包含在连接子之后立即开始并且就在β链恒定区开始之前结束的序列。全长α链(seqidno:325)包含从氨基末端开始并且就在连接子开始之前结束的序列。全长β链(seqidno:326)包含在连接子之后立即开始并且以羧基末端结束的序列。癌症反应性t细胞使用下面列出的筛选方法进行鉴定。用于分离tcr的方法示于下文。tcr名称:4213-f2-tcr1p53突变的识别:r248q筛选方法:p53“热点”突变通用筛选共培养以鉴定tcr:共培养4213-f2与r248q长肽，分选cd8+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:8.3％(在24对中观察到2次)tcr取向:α-β表达载体:sb转座子患者4213的tcr4213-f2-tcr1的统计数据示于下表21中。表21参数#频率总孔96100％cdr3α33.1％cdr3β88.3％4213-f2-tcr1对22.1％总配对的tcr2425.0％从患者4213分离的4213-f2-tcr2的序列示于下文。从氨基末端开始，第一加下划线区是cdr1α(seqidno:327)，第二加下划线区是cdr2α(seqidno:328)，第三加下划线区是cdr3α(seqidno:329)，第四加下划线区是cdr1β(seqidno:330)，第五加下划线区是cdr2β(seqidno:331)，和第六加下划线区是cdr3β(seqidno:332)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一加斜体区是α链恒定区(seqidno:23)并且第二加斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:333)包含从氨基末端开始并且就在α链恒定区的开始之前结束的序列。β链可变区(seqidno:334)包含在连接子之后立即开始并且就在β链恒定区开始之前结束的序列。全长α链(seqidno:335)包含从氨基末端开始并且就在连接子开始之前结束的序列。全长β链(seqidno:336)包含在连接子之后立即开始并且以羧基末端结束的序列。癌症反应性t细胞使用下面列出的筛选方法进行鉴定。用于分离tcr的方法示于下文。tcr名称:4213-f2-tcr2p53突变的识别:r248q筛选方法:p53“热点”突变通用筛选共培养以鉴定tcr:共培养4213-f2与r248q长肽，分选cd8+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:12.5％(在24对中观察到3次)tcr取向:α-β表达载体:sb转座子患者4213的tcr4213-f2-tcr2的统计数据示于下表22中。表22参数#频率总孔96100％cdr3α44.2％cdr3β44.2％4213-f2-tcr2对33.1％总配对的tcr2425.0％从患者4213分离的4213-f2-tcr3的序列示于下文。从氨基末端开始，第一加下划线区是cdr1α(seqidno:337)，第二加下划线区是cdr2α(seqidno:338)，第三加下划线区是cdr3α(seqidno:339)，第四加下划线区是cdr1β(seqidno:340)，第五加下划线区是cdr2β(seqidno:341)，并且第六加下划线区是cdr3β(seqidno:342)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一加斜体区是α链恒定区(seqidno:23)并且第二加斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:343)包含从氨基末端开始并且就在α链恒定区的开始之前结束的序列。β链可变区(seqidno:344)包含在连接子之后立即开始并且就在β链恒定区开始之前结束的序列。全长α链(seqidno:345)包含从氨基末端开始并且就在连接子开始之前结束的序列。全长β链(seqidno:346)包含在连接子之后立即开始并且以羧基末端结束的序列。癌症反应性t细胞使用下面列出的筛选方法进行鉴定。用于分离tcr的方法示于下文。tcr名称:4213-f2-tcr3p53突变的识别:r248q筛选方法:p53“热点”突变通用筛选共培养以鉴定tcr:共培养4213-f2与r248q长肽，分选cd8+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:70.8％(在24对中观察到17次)tcr取向:α-β表达载体:sb转座子患者4213的tcr4213-f2-tcr3的统计数据示于下表23中。表23参数#频率总孔96100％cdr3α1919.8％cdr3β4243.8％4213-f2-tcr3对1717.7％总配对的tcr2425.0％从患者4213分离的4213-f24-tcra1的序列示于下文。从氨基末端开始，第一加下划线区是cdr1α(seqidno:347)，第二加下划线区是cdr2α(seqidno:348)，第三加下划线区是cdr3α(seqidno:349)，第四加下划线区是cdr1β(seqidno:350)，第五加下划线区是cdr2β(seqidno:351)，和第六加下划线区是cdr3β(seqidno:352)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一加斜体区是α链恒定区(seqidno:23)并且第二加斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:353)包含从氨基末端开始并且就在α链恒定区的开始之前结束的序列。β链可变区(seqidno:354)包含在连接子之后立即开始并且就在β链恒定区开始之前结束的序列。全长α链(seqidno:355)包含从氨基末端开始并且就在连接子开始之前结束的序列。全长β链(seqidno:356)包含在连接子之后立即开始并且以羧基末端结束的序列。癌症反应性t细胞使用下面列出的筛选方法进行鉴定。用于分离tcr的方法示于下文。tcr名称:4213-f24-tcra1p53突变的识别:r248q筛选方法:p53“热点”突变通用筛选共培养以鉴定tcr:共培养4213-f24与r248q长肽，分选cd8+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:15.9％(在44对中观察到7次)tcr取向:α-β表达载体:sb转座子患者4213的tcr4213-f24-tcra1的统计数据示于下表24中。表24参数#频率总孔96100％cdr3α77.3％cdr3β8083.3％4213-f24-tcra1对77.3％总配对的tcr4445.8％从患者4213分离的4213-f24-tcra2的序列示于下文。从氨基末端开始，第一加下划线区是cdr1α(seqidno:357)，第二加下划线区是cdr2α(seqidno:358)，第三加下划线区是cdr3α(seqidno:359)，第四加下划线区是cdr1β(seqidno:360)，第五加下划线区是cdr2β(seqidno:361)，和第六加下划线区是cdr3β(seqidno:362)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一加斜体区是α链恒定区(seqidno:23)并且第二加斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:363)包含从氨基末端开始并且就在α链恒定区开始之前结束的序列。β链可变区(seqidno:364)包含在连接子之后立即开始并且就在β链恒定区开始之前结束的序列。全长α链(seqidno:365)包含从氨基末端开始并且就在连接子开始之前结束的序列。全长β链(seqidno:366)包含在连接子之后立即开始并且以羧基末端结束的序列。癌症反应性t细胞使用下面列出的筛选方法进行鉴定。用于分离tcr的方法示于下文。tcr名称:4213-f24-tcra2p53突变的识别:r248q筛选方法:p53“热点”突变通用筛选共培养以鉴定tcr:共培养4213-f24与r248q长肽，分选cd8+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:84.1％(在44对中观察到37次)tcr取向:α-β表达载体:sb转座子患者4213的tcr4213-f24-tcra2的统计数据示于下表25中。表25参数#频率总孔96100％cdr3α3940.6％cdr3β8083.3％4213-f24-tcra2对3738.5％总配对的tcr4445.8％从患者4213分离的4213-pbl-tcr1的序列示于下文。从氨基末端开始，第一加下划线区是cdr1α(seqidno:367)，第二加下划线区是cdr2α(seqidno:368)，第三加下划线区是cdr3α(seqidno:369)，第四加下划线区是cdr1β(seqidno:370)，第五加下划线区是cdr2β(seqidno:371)，和第六加下划线区是cdr3β(seqidno:372)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一加斜体区是α链恒定区(seqidno:23)并且第二加斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:373)包含从氨基末端开始并且就在α链恒定区开始之前结束的序列。β链可变区(seqidno:374)包含在连接子之后立即开始并且就在β链恒定区开始之前结束的序列。全长α链(seqidno:375)包含从氨基末端开始并且就在连接子开始之前结束的序列。全长β链(seqidno:376)包含在连接子之后立即开始并且以羧基末端结束的序列。癌症反应性t细胞使用下面列出的筛选方法进行鉴定。用于分离tcr的方法示于下文。tcr名称:4213-pbl-tcr1p53突变的识别:r248q筛选方法:p53“热点”突变通用筛选共培养以鉴定tcr:将在用r248q长肽体外致敏之后的cd4+记忆t细胞与r248q长肽共培养，并分选cd4+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:9.5％(在63对中观察到6次)tcr取向:α-β表达载体:sb转座子患者4213的tcr4213-pbl-tcr1的统计数据示于下表26中。表26从患者4213分离的4213-pbl-tcr2的序列示于下文。从氨基末端开始，第一加下划线区是cdr1α(seqidno:377)，第二加下划线区是cdr2α(seqidno:378)，第三加下划线区是cdr3α(seqidno:379)，第四加下划线区是cdr1β(seqidno:380)，第五加下划线区是cdr2β(seqidno:381)，和第六加下划线区是cdr3β(seqidno:382)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一加斜体区是α链恒定区(seqidno:23)并且第二加斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:383)包含从氨基末端开始并且在α链恒定区开始之前结束的序列。β链可变区(seqidno:384)包含在连接子之后立即开始并且就在β链恒定区开始之前结束的序列。全长α链(seqidno:385)包含从氨基末端开始并且就在连接子开始之前结束的序列。全长β链(seqidno:386)包含在连接子之后立即开始并且以羧基末端结束的序列.癌症反应性t细胞使用下面列出的筛选方法进行鉴定。用于分离tcr的方法示于下文。tcr名称:4213-pbl-tcr2p53突变的识别:r248q筛选方法:p53“热点”突变通用筛选共培养以鉴定tcr:将在用r248q长肽体外致敏之后的cd4+记忆t细胞与r248q长肽共培养，并分选cd4+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:7.9％(在63对中观察到5次)tcr取向:α-β表达载体:sb转座子患者4213的tcr4213-pbl-tcr2的统计数据示于下表27中。表27参数#频率总孔192100％cdr3α84.2％cdr3β84.2％4213-pbl-tcr2对52.6％总配对的tcr6332.8％从患者4213分离的4213-pbl-tcr3的序列示于下文。从氨基末端开始，第一加下划线区是cdr1α(seqidno:387)，第二加下划线区是cdr2α(seqidno:388)，第三加下划线区是cdr3α(seqidno:389)，第四加下划线区是cdr1β(seqidno:390)，第五加下划线区是cdr2β(seqidno:391)，和第六加下划线区是cdr3β(seqidno:392)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一加斜体区是α链恒定区(seqidno:23)并且第二加斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:393)包含从氨基末端开始并且就在α链恒定区开始之前结束的序列。β链可变区(seqidno:394)包含在连接子之后立即开始并且就在β链恒定区开始之前结束的序列。全长α链(seqidno:395)包含从氨基末端开始并且就在连接子开始之前结束的序列。全长β链(seqidno:396)包含在连接子之后立即开始并且以羧基末端结束的序列。癌症反应性t细胞使用下面列出的筛选方法进行鉴定。用于分离tcr的方法示于下文。tcr名称:4213-pbl-tcr3p53突变的识别:r248q筛选方法:p53“热点”突变通用筛选共培养以鉴定tcr:将在用r248q长肽体外致敏之后的cd4+记忆t细胞与r248q长肽共培养，并分选cd4+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:6.3％(在63对中观察到4次)tcr取向:α-β表达载体:sb转座子患者4213的tcr4213-pbl-tcr3的统计数据示于下表28中。表28从患者4213分离的4213-pbl-tcr4a1的序列示于下文。从氨基末端开始，第一加下划线区是cdr1α(seqidno:397)，第二加下划线区是cdr2α(seqidno:398)，第三加下划线区是cdr3α(seqidno:399)，第四加下划线区是cdr1β(seqidno:400)，第五加下划线区是cdr2β(seqidno:401)，和第六加下划线区是cdr3β(seqidno:402)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一加斜体区是α链恒定区(seqidno:23)并且第二加斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:403)包含从氨基末端开始并且就在α链恒定区的开始之前结束的序列。β链可变区(seqidno:404)包含在连接子之后立即开始并且就在β链恒定区开始之前结束的序列。全长α链(seqidno:405)包含从氨基末端开始并且就在连接子开始之前结束的序列。全长β链(seqidno:406)包含在连接子之后立即开始并且以羧基末端结束的序列。癌症反应性t细胞使用下面列出的筛选方法进行鉴定。用于分离tcr的方法示于下文。tcr名称:4213-pbl-tcr4a1p53突变的识别:r248q筛选方法:p53“热点”突变通用筛选共培养以鉴定tcr:将在用r248q长肽体外致敏之后的cd4+记忆t细胞与r248q长肽共培养并分选cd4+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:3.2％(在63对中观察到2次)tcr取向:α-β表达载体:sb转座子患者4213的tcr4213-pbl-tcr4a1的统计数据示于下表29中。表29参数#频率总孔192100％cdr3α21.0％cdr3β3618.8％4213-pbl-tcr4a1对21.0％总配对的tcr6332.8％从患者4213分离的4213-pbl-tcr4a2的序列示于下文。从氨基末端开始，第一加下划线区是cdr1α(seqidno:407)，第二加下划线区是cdr2α(seqidno:408)，第三加下划线区是cdr3α(seqidno:409)，第四加下划线区是cdr1β(seqidno:410)，第五加下划线区是cdr2β(seqidno:411)，和第六加下划线区是cdr3β(seqidno:412)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一加斜体区是α链恒定区(seqidno:23)并且第二加斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:413)包含从氨基末端开始并且就在α链恒定区开始之前结束的序列。β链可变区(seqidno:414)包含在连接子之后立即开始并且就在β链恒定区开始之前结束的序列。全长α链(seqidno:415)包含从氨基末端开始并且就在连接子开始之前结束的序列。全长β链(seqidno:416)包含在连接子之后立即开始并且以羧基末端结束的序列。癌症反应性t细胞使用下面列出的筛选方法进行鉴定。用于分离tcr的方法示于下文。tcr名称:4213-pbl-tcr4a2p53突变的识别:r248q筛选方法:p53“热点”突变通用筛选共培养以鉴定tcr:将在用r248q长肽体外致敏之后的cd4+记忆t细胞与r248q长肽共培养并分选cd4+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:3.2％(在63对中观察到2次)tcr取向:α-β表达载体:sb转座子患者4213的tcr4213-pbl-tcr4a2的统计数据示于下表30中。表30参数#频率总孔192100％cdr3α21.0％cdr3β3618.8％4213-pbl-tcr4a2对21.0％总配对的tcr6332.8％从患者4213分离的4213-pbl-tcr4a3的序列示于下文。从氨基末端开始，第一加下划线区是cdr1α(seqidno:417)，第二加下划线区是cdr2α(seqidno:418)，第三加下划线区是cdr3α(seqidno:419)，第四加下划线区是cdr1β(seqidno:420)，第五加下划线区是cdr2β(seqidno:421)，和第六加下划线区是cdr3β(seqidno:422)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一加斜体区是α链恒定区(seqidno:23)并且第二加斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:423)包含从氨基末端开始并且就在α链恒定区开始之前结束的序列。β链可变区(seqidno:424)包含在连接子之后立即开始并且就在β链恒定区开始之前结束的序列。全长α链(seqidno:425)包含从氨基末端开始并且就在连接子开始之前结束的序列。全长β链(seqidno:426)包含在连接子之后立即开始并且以羧基末端结束的序列。癌症反应性t细胞使用下面列出的筛选方法进行鉴定。用于分离tcr的方法示于下文。tcr名称:4213-pbl-tcr4a3p53突变的识别:r248q筛选方法:p53“热点”突变通用筛选共培养以鉴定tcr:将在用r248q长肽体外致敏之后的cd4+记忆t细胞与r248q长肽共培养并分选cd4+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:4.8％(在63对中观察到3次)tcr取向:α-β表达载体:sb转座子患者4213的tcr4213-pbl-tcr4a3的统计数据示于下表31中。表31参数#频率总孔192100％cdr3α42.1％cdr3β3618.8％4213-pbl-tcr4a3对31.6％总配对的tcr6332.8％从患者4213分离的4213-pbl-tcr4a4的序列示于下文。从氨基末端开始，第一加下划线区是cdr1α(seqidno:427)，第二加下划线区是cdr2α(seqidno:428)，第三加下划线区是cdr3α(seqidno:429)，第四加下划线区是cdr1β(seqidno:430)，第五加下划线区是cdr2β(seqidno:431)，和第六加下划线区是cdr3β(seqidno:432)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一加斜体区是α链恒定区(seqidno:23)并且第二加斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:433)包含从氨基末端开始并且就在α链恒定区开始之前结束的序列。β链可变区(seqidno:434)包含在连接子之后立即开始并且就在β链恒定区开始之前结束的序列。全长α链(seqidno:435)包含从氨基末端开始并且就在连接子开始之前结束的序列。全长β链(seqidno:436)包含在连接子之后立即开始并且以羧基末端结束的序列。癌症反应性t细胞使用下面列出的筛选方法进行鉴定。用于分离tcr的方法示于下文。tcr名称:4213-pbl-tcr4a4p53突变的识别:r248q筛选方法:p53“热点”突变通用筛选共培养以鉴定tcr:将在用r248q长肽体外致敏之后的cd4+记忆t细胞与r248q长肽共培养并分选cd4+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:3.2％(在63对中观察到2次)tcr取向:α-β表达载体:sb转座子患者4213的tcr4213-pbl-tcr4a4的统计数据示于下表32中。表32参数#频率总孔192100％cdr3α21.0％cdr3β3618.8％4213-pbl-tcr4a3对21.0％总配对的tcr6332.8％实施例8该实施例证实了通过在自体t细胞内共培养被诱导以表达突变的p53的自体apc来鉴定患者4268中的抗-突变的p53t细胞(“p53热点突变通用筛选”)。该实施例还证实了从患者4268分离五种抗-突变的p53tcr。如针对患者4268的图27-30所述进行实验。将来自患者4268的til片段(f1-f24，n＝24)与用由无关、wtp53或经突变的p53序列组成的tmg电穿孔的自体apc共培养。在37℃下进行共培养过夜。使用elispot测定评价ifn-γ的分泌。结果示于图27中。将来自患者4268的til片段(f1-f24，n＝24)与用肽媒介物(dmso)或由wtp53-r248序列或经突变的p53-r248q序列组成的经纯化的(>95％，通过hplc)25-个氨基酸的肽脉冲的自体apc共培养。在37℃下进行共培养过夜。使用elispot测定评价ifn-γ的分泌。结果示于图28中。将来自患者4268的til片段(f1-f24，n＝24)与用肽媒介物(dmso)或由wtp53-r248序列或经突变的p53-r248q序列组成的经纯化的(>95％，通过hplc)25-个氨基酸的肽脉冲的自体apc共培养。在37℃下进行共培养过夜。对于淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)设门后，通过流式细胞术评价4-1bb的表达。结果示于图29中。til片段f18和f19是对cd4+41bb+t细胞进行分选后的tcr的来源。将来自患者4268的til片段(f1-f24，n＝24)与用肽媒介物(dmso)或由wtp53-r248序列或经突变的p53-r248q序列组成的经纯化的(>95％，通过hplc)25-个氨基酸的肽脉冲的自体apc共培养。在37℃下进行共培养过夜。对于淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)设门后，通过流式细胞术评价4-1bb的表达。结果示于图30中。til片段f7,f8和f15是对cd8+4-1bb+t细胞进行分选后的tcr的来源。从患者4268分离的4268-tcr1的序列示于下文。从氨基末端开始，第一加下划线区是cdr1α(seqidno:137)，第二加下划线区是cdr2α(seqidno:138)，第三加下划线区是cdr3α(seqidno:139)，第四加下划线区是cdr1β(seqidno:140)，第五加下划线区是cdr2β(seqidno:141)，和第六加下划线区是cdr3β(seqidno:142)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一加斜体区是α链恒定区(seqidno:23)并且第二加斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:143)包含从氨基末端开始并且在α链恒定区开始之前就结束的序列。β链可变区(seqidno:144)包含在连接子之后立即开始并且在β链恒定区开始之前就结束的序列。全长α链(seqidno:145)包含从氨基末端开始并且在连接子开始之前就结束的序列。全长β链(seqidno:146)包含在连接子之后立即开始并且以羧基末端结束的序列。癌症反应性t细胞使用下面列出的筛选方法进行鉴定。用于分离tcr的方法示于下文。tcr名称:4268-tcr1p53突变的识别:r248q方法:p53“热点”突变通用筛选共培养以鉴定tcr:共培养4268-f7和4268-f8与p53-r248q长肽，分选cd8+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:78.7％(在61对中观察到48次)tcr取向:α-β表达载体:sb转座子患者4268的tcr4268-tcr1的统计数据示于下表33中。表33从患者4268分离的4268-tcr2的序列示于下文。从氨基末端开始，第一加下划线区是cdr1α(seqidno:147)，第二加下划线区是cdr2α(seqidno:148)，第三加下划线区是cdr3α(seqidno:149)，第四加下划线区是cdr1β(seqidno:150)，第五加下划线区是cdr2β(seqidno:151)，和第六加下划线区是cdr3β(seqidno:152)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一加斜体区是α链恒定区(seqidno:23)并且第二加斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:153)包含从氨基末端开始并且在α链恒定区开始之前就结束的序列。β链可变区(seqidno:154)包含在连接子之后立即开始并且就β链恒定区开始之前结束的序列。全长α链(seqidno:155)包含从氨基末端开始并且就在连接子开始之前结束的序列。全长β链(seqidno:156)包含在连接子之后立即开始并且以羧基末端结束的序列。癌症反应性t细胞使用下面列出的筛选方法进行鉴定。用于分离tcr的方法示于下文。tcr名称:4268-tcr2p53突变的识别:r248q方法:p53“热点”突变通用筛选共培养以鉴定tcr:共培养4268-f7和4268-f8与p53-r248q长肽，分选cd8+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:6.6％(在61对中观察到4次)tcr取向:α-β表达载体:sb转座子患者4268的tcr4268-tcr2的统计数据示于下表34中。表34参数#频率总孔96100％cdr3α66.3％cdr3β44.2％4268-tcr2对44.2％总配对的tcr6163.5％从患者4268分离的4268-tcr3的序列示于下文。从氨基末端开始，第一加下划线区是cdr1α(seqidno:157)，第二加下划线区是cdr2α(seqidno:158)，第三加下划线区是cdr3α(seqidno:159)，第四加下划线区是cdr1β(seqidno:160)，第五加下划线区是cdr2β(seqidno:161)，和第六加下划线区是cdr3β(seqidno:162)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一加斜体区是α链恒定区(seqidno:23)并且第二加斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:163)包含从氨基末端开始并且在α链恒定区开始之前就结束的序列。β链可变区(seqidno:164)包含在连接子之后立即开始并且就在β链恒定区开始之前结束的序列。全长α链(seqidno:165)包含从氨基末端开始并且就在连接子开始之前结束的序列。全长β链(seqidno:166)包含在连接子之后立即开始并且以羧基末端结束的序列。癌症反应性t细胞使用下面列出的筛选方法进行鉴定。用于分离tcr的方法示于下文。tcr名称:4268-tcr3p53突变的识别:r248q方法:p53“热点”突变通用筛选共培养以鉴定tcr:共培养4268-f15与p53-r248q长肽，分选cd8+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:88.6％(在35对中观察到31次)tcr取向:α-β表达载体:sb转座子患者4268的tcr4268-tcr3的统计数据示于下表35中。表35参数#频率总孔96100％cdr3α4243.8％cdr3β3738.5％4268-tcr3对3132.3％总配对的tcr3536.5％从患者4268分离的4268-tcr4的序列示于下文。从氨基末端开始，第一加下划线区是cdr1α(seqidno:167)，第二加下划线区是cdr2α(seqidno:168)，第三加下划线区是cdr3α(seqidno:169)，第四加下划线区是cdr1β(seqidno:170)，第五加下划线区是cdr2β(seqidno:171)，和第六加下划线区是cdr3β(seqidno:172)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一加斜体区是α链恒定区(seqidno:23)并且第二加斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:173)包含从氨基末端开始并且就在α链恒定区开始之前结束的序列。β链可变区(seqidno:174)包含在连接子之后立即开始并且就在β链恒定区开始之前结束的序列。全长α链(seqidno:175)包含从氨基末端开始并且在连接子开始之前就结束的序列。全长β链(seqidno:176)包含在连接子之后立即开始并且以羧基末端结束的序列。癌症反应性t细胞使用下面列出的筛选方法进行鉴定。用于分离tcr的方法示于下文。tcr名称:4268-tcr4p53突变的识别:r248q方法:p53“热点”突变通用筛选共培养以鉴定tcr:共培养4268-f18与p53-r248q长肽，分选cd4+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:95.2％(在42对中观察到40次)tcr取向:α-β表达载体:sb转座子患者4268的tcr4268-tcr4的统计数据示于下表36中。表36参数#频率总孔96100％cdr3α4344.8％cdr3β5355.2％4268-tcr4对4041.7％总配对的tcr4243.8％从患者4268分离的4268-tcr5的序列示于下文。从氨基末端开始，第一加下划线区是cdr1α(seqidno:177)，第二加下划线区是cdr2α(seqidno:178)，第三加下划线区是cdr3α(seqidno:179)，第四加下划线区是cdr1β(seqidno:180)，第五加下划线区是cdr2β(seqidno:181)，和第六加下划线区是cdr3β(seqidno:182)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一加斜体区是α链恒定区(seqidno:23)并且第二加斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:183)包含从氨基末端开始并且在α链恒定区开始之前就结束的序列。β链可变区(seqidno:184)包含在连接子之后立即开始并且就在β链恒定区开始之前结束的序列。全长α链(seqidno:185)包含从氨基末端开始并且就在连接子开始之前结束的序列。全长β链(seqidno:186)包含在连接子之后立即开始并且以羧基末端结束的序列。癌症反应性t细胞使用下面列出的筛选方法进行鉴定。用于分离tcr的方法示于下文。tcr名称:4268-tcr5p53突变的识别:r248q方法:p53“热点”突变通用筛选共培养以鉴定tcr:共培养4268-f19与p53-mut-tmg，分选cd4+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:11.8％(在17对中观察到2次)tcr取向:α-β表达载体:sb转座子患者4268的tcr4268-tcr5的统计数据示于下表37中。表37参数#频率总孔96100％cdr3α22.1％cdr3β8790.6％4268-tcr5对22.1％总配对的tcr1717.7％实施例9该实施例证实了通过在自体t细胞内共培养被诱导以表达突变的p53的自体apc来鉴定患者4266中的抗-突变的p53t细胞(“p53热点突变通用筛选”)。该实施例还证实了从患者4266分离四种抗-突变的p53tcr。如图31-34和53-55针对患者4266所述进行实验。将来自患者4266的til片段(f1-f24，n＝24)与用由无关、wtp53或经突变的p53序列组成的tmg电穿孔的自体apc共培养。在37℃下进行共培养过夜。使用elispot测定评价ifn-γ的分泌。结果显示于图31中。对于淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)设门后，通过流式细胞术评价4-1bb的表达。结果显示于图32中。将来自患者4266的til片段(f1-f24，n＝24)与用肽媒介物(dmso)或由wtp53-r248序列或经突变的p53-r248w序列组成的经纯化的(>95％，通过hplc)25-个氨基酸的肽脉冲的自体apc共培养。在37℃下进行共培养过夜。使用elispot测定评价ifn-γ的分泌。结果显示于图33中。对于淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)设门后，通过流式细胞术评价4-1bb的表达。结果显示于图34中。将cos7细胞(每孔2.5x104个)涂铺在平底96孔板的孔上。第二天，将细胞与来自患者4266的单独hla等位基因共转染。次日，将细胞用无肽、dmso、野生型p53-r248肽sscmggmnrr(seqidno:590)或经突变的p53-r248w肽sscmggmnwr(seqidno:591)在37℃下以1μg/ml脉冲2小时。将来自患者4266的til培养物(105)添加至孔并在37℃下共培养过夜。对于淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)→cd4-cd8+设门后，通过流式细胞术评价4-1bb的表达。结果显示于图53中。将对从4266-til鉴定的p53-r248w具有假定特异性的表达模拟物(无tcr)、4266-tcr1、4266-tcr2、4266-tcr3或4266-tcr4的t细胞与用肽媒介物(dmso)或由wtp53-r248序列或经突变的p53-r248w序列组成的经纯化的(>95％，通过hplc)25个氨基酸的肽脉冲的自体apc共培养。单独的培养基及pma和离子霉素分别为阴性对照和阳性对照。在37℃下进行共培养过夜。对于淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)→cd4-cd8+设门后，通过流式细胞术评价4-1bb的表达。结果显示于图54中。肿瘤细胞(tc)系由从患者4266切除的异种移植的肿瘤碎片建立，然后通过免疫受损的小鼠连续传代(tc#4266)。将tc#4266与表达模拟物(无tcr)或p53-r248w-特异性tcr(4266-tcr2、4266-tcr3或4266-tcr4)的t细胞(105)在37℃下共培养过夜。将tc#4266细胞不与任何东西孵育、或者将tc#4266细胞与w6/32泛-hlai类特异性阻断抗体、iva12泛-hlaii类特异性阻断抗体或突变的p53-r248w肽sscmggmnwr(seqidno:591)在37℃下孵育2小时。将抗体以5μg/ml的终浓度保存在共培养物中。将肽以1μg/ml孵育，并且孵育后洗涤过量肽。单独的培养基(无tc)及pma和离子霉素分别为阴性对照和阳性对照。对于淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)→cd4-cd8+设门后，通过流式细胞术评价4-1bb的表达。结果示于图55中。从患者4266分离的4266-tcr1的序列示于下文。从氨基末端开始，第一加下划线区是cdr1α(seqidno:97)，第二加下划线区是cdr2α(seqidno:98)，第三加下划线区是cdr3α(seqidno:99)，第四加下划线区是cdr1β(seqidno:100)，第五加下划线区是cdr2β(seqidno:101)，和第六加下划线区是cdr3β(seqidno:102)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一加斜体区是α链恒定区(seqidno:23)并且第二加斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:103)包含从氨基末端开始并且在α链恒定区开始之前就结束的序列。β链可变区(seqidno:104)包含在连接子之后立即开始并且就在β链恒定区开始之前结束的序列。全长α链(seqidno:105)包含从氨基末端开始并且就在连接子开始之前结束的序列。全长β链(seqidno:106)包含在连接子之后立即开始并且以羧基末端结束的序列。癌症反应性t细胞使用下面列出的筛选方法进行鉴定。用于分离tcr的方法示于下文。tcr名称:4266-tcr1p53突变的识别:r248w筛选方法:p53“热点”突变通用筛选共培养以鉴定tcr:共培养4266-f1、4266-f3、4266-f5和4266-f6与p53muttmg或r248w长肽(两次共培养均检测相同的tcr)，分选cd8+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:17.0％(在53对中观察到9次)tcr取向:α-β表达载体:sb转座子患者4266的tcr4266-tcr1的统计数据示于下表38中。表38参数#频率总孔96100％cdr3α99.4％cdr3β1010.4％4266-tcr1对99.4％总配对的tcr5355.2％从患者4266分离的4266-tcr2的序列示于下文。从氨基末端开始，第一加下划线区是cdr1α(seqidno:107)，第二加下划线区是cdr2α(seqidno:108)，第三加下划线区是cdr3α(seqidno:109)，第四加下划线区是cdr1β(seqidno:110)，第五加下划线区是cdr2β(seqidno:111)，和第六加下划线区是cdr3β(seqidno:112)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一加斜体区是α链恒定区(seqidno:23)并且第二加斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:113)包含从氨基末端开始并且在α链恒定区开始之前就结束的序列。β链可变区(seqidno:114)包含在连接子之后立即开始并且在β链恒定区开始之前就结束的序列。全长α链(seqidno:115)包含从氨基末端开始并且在连接子开始之前就结束的序列。全长β链(seqidno:116)包含在连接子之后立即开始并且以羧基末端结束的序列。癌症反应性t细胞使用下面列出的筛选方法进行鉴定。用于分离tcr的方法示于下文。tcr名称:4266-tcr2p53突变的识别:r248w筛选方法:p53“热点”突变通用筛选共培养以鉴定tcr:共培养4266-f1、4266-f3、4266-f5和4266-f6与p53muttmg或r248w长肽(两次共培养均检测相同的tcr)，分选cd8+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:24.5％(在53对中观察到13次)tcr取向:α-β表达载体:sb转座子患者4266的tcr4266-tcr2的统计数据示于下表39中。表39参数#频率总孔96100％cdr3α2121.9％cdr3β1818.8％4266-tcr2对1313.5％总配对的tcr5355.2％从患者4266分离的4266-tcr3的序列示于下文。从氨基末端开始，第一加下划线区是cdr1α(seqidno:117)，第二加下划线区是cdr2α(seqidno:118)，第三加下划线区是cdr3α(seqidno:119)，第四加下划线区是cdr1β(seqidno:120)，第五加下划线区是cdr2β(seqidno:121)，和第六加下划线区是cdr3β(seqidno:122)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一加斜体区是α链恒定区(seqidno:23)并且第二加斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:123)包含从氨基末端开始并且在α链恒定区开始之前就结束的序列。β链可变区(seqidno:124)包含在连接子之后立即开始并且在β链恒定区开始之前就结束的序列。全长α链(seqidno:125)包含从氨基末端开始并且就在连接子开始之前结束的序列。全长β链(seqidno:126)包含在连接子之后立即开始并且以羧基末端结束的序列。癌症反应性t细胞使用下面列出的筛选方法进行鉴定。用于分离tcr的方法示于下文。tcr名称:4266-tcr3p53突变的识别:r248w筛选方法:p53“热点”突变通用筛选共培养以鉴定tcr:共培养4266-f1、4266-f3、4266-f5和4266-f6与p53muttmg或r248w长肽(两次共培养均检测相同的tcr)，分选cd8+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:34.0％(在53对中观察到18次)tcr取向:α-β表达载体:sb转座子患者4266的tcr4266-tcr3的统计数据示于下表40中。表40参数#频率总孔96100％cdr3α1919.8％cdr3β2627.14266-tcr3对1818.8％总配对的tcr5355.2％从患者4266分离的4266-tcr4的序列示于下文。从氨基末端开始，第一加下划线区是cdr1α(seqidno:127)，第二加下划线区是cdr2α(seqidno:128)，第三加下划线区是cdr3α(seqidno:129)，第四加下划线区是cdr1β(seqidno:130)，第五加下划线区是cdr2β(seqidno:131)，和第六加下划线区是cdr3β(seqidno:132)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一加斜体区是α链恒定区(seqidno:23)并且第二加斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:133)包含从氨基末端开始并且就在α链恒定区开始之前结束的序列。β链可变区(seqidno:134)包含在连接子之后立即开始并且就在β链恒定区开始之前结束的序列。全长α链(seqidno:135)包含从氨基末端开始并且就在连接子开始之前结束的序列。全长β链(seqidno:136)包含在连接子之后立即开始并且以羧基末端结束的序列。癌症反应性t细胞使用下面列出的筛选方法进行鉴定。用于分离tcr的方法示于下文。tcr名称:4266-tcr4p53突变的识别:r248w筛选方法:p53“热点”突变通用筛选共培养以鉴定tcr:共培养4266-f1、4266-f3、4266-f5和4266-f6与p53muttmg或r248w长肽(两次共培养均检测相同的tcr)，分选cd8+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:9.4％(在53对中观察到5次)tcr取向:α-β表达载体:sb转座子患者4266的tcr4266-tcr4的统计数据示于下表41中。表41参数#频率总孔96100％cdr3α55.2％cdr3β1313.54266-tcr3对55.2％总配对的tcr5355.2％实施例10该实施例证实了t细胞对p53“热点”突变的响应的概述。表42提供了t细胞对p53“热点”突变的响应的概述。表42的第1-15号是一项回顾性研究。表42的第16-33号是一项前瞻性研究。表42n/a＝不适用；n＝阴性；y＝确认有反应性；tbs＝待筛选。实施例11该实施例证实了用p53突变-反应性til治疗患者。表43提供了用p53突变-反应性til治疗患者的概述。表43nt＝未治疗；tbs＝待筛选；n.r.＝无响应；p.r.＝部分响应；**患者尚未治疗。实施例12该实施例证实了来自患者4141的tcr的分离和特异的反应性。将自体apc用编码无关突变、wtp53序列或包含r175h的突变的p53序列的tmg转染。单独的培养基以及pma和离子霉素分别为阴性对照和阳性对照。第二天，将来自患者4141的til(片段培养物12)与tmg-转染的apc在37℃下共培养过夜。通过elispot评价ifn-γ的分泌。对于淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)→cd4-cd8+设门后，通过流式细胞术评价4-1bb的表达。结果示于图38中。将cos7细胞(每孔2.5x104个)涂铺在平底96孔板的孔上。第二天，将细胞用来自患者4141的单独hla等位基因以及无额外基因、wttp53tmg或含有p53-r175h序列的突变的tp53tmg共转染。第二天，将来自患者4141的对p53-r175h具有特异性的til(片段培养物12)与转染的cos7细胞共培养，并在37℃下孵育过夜。通过elispot评价ifn-γ的分泌。结果示于图39中。将表达模拟物(无tcr)或4141-tcr1a2的t细胞与t2肿瘤细胞(表达hla-a*02:01)共培养。将t2细胞在37℃下用肽媒介物(dmso)或由wtp53-r175肽hmtevvrrc(seqidno:532)或经突变的p53-r175h肽hmtevvrhc(seqidno:530)组成的纯化的(>95％，通过hplc)肽脉冲。单独的培养基及pma和离子霉素分别为阴性对照和阳性对照。在37℃下进行共培养过夜。ifn-γ的分泌通过elisa评价。结果示于图40中。将表达4141-tcr1a2的t细胞与saos2细胞(p53-null和hla-a*02:01+)在37℃下共培养过夜，这些细胞未经处理，或使得过表达全长p53-r175h蛋白。将分泌的抑制剂(莫能菌素和布雷菲德菌素a)添加至共培养物，以捕获t细胞内的细胞因子。共培养6小时后，将细胞固定并透化，然后针对il-2、cd107a、ifn-γ和肿瘤坏死因子-α(tnfα)染色。流式细胞术用于基于淋巴细胞设门分析共培养物。结果示于图41中。从患者4141分离的tcr4141-tcr1a2的序列示于以下。从氨基末端开始，第一加下划线的区是cdr1α(seqidno:467)，第二加下划线的区是cdr2α(seqidno:468)，第三加下划线的区是cdr3α(seqidno:469)，第四加下划线的区是cdr1β(seqidno:470)，第五加下划线的区是cdr2β(seqidno:471)，并且第六加下划线的区是cdr3β(seqidno:472)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一斜体区是α链恒定区(seqidno:23)并且第二斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:473)包含从氨基末端开始并且就在α链恒定区开始前结束的序列。β链可变区(seqidno:474)包含在连接子之后立即开始并且就在β链恒定区开始之前结束的序列。全长α链(seqidno:475)包含从氨基末端开始并且就在连接子开始之前结束的序列。全长β链(seqidno:476)包含在连接子之后立即开始并且以羧基末端结束的序列。如下所述鉴定癌症反应性t细胞。如下所述分离tcr。tcr名称:4141-tcr1a2p53突变的识别:r175h筛选方法:p53“热点”突变通用筛选共培养以鉴定tcr:共培养4141输液袋til与p53muttmg，和分选cd8+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr，然后tatopo克隆试剂盒(thermofisherscientific,waltham,ma)用于α链tcr取向:α-β表达载体:sb转座子来自患者4141的4141-tcr1a2的统计数据示于下表44中。表44参数#频率总孔96100％cdr3α未知(tatopo克隆)不适用cdr3β5860.4％实施例13该实施例证实了从患者4259分离的tcr的分离和特异的反应性。将来自患者4259的til片段培养物(no.6)与自体apc共培养，所述自体apc用1)由无关的、wtp53或突变的p53序列组成的tmg电穿孔或者2)用肽媒介物(dmso)或由wtp53-y220序列或经突变的p53-y220序列组成的纯化的(>95％，通过hplc)25个氨基酸的肽脉冲。在37℃下进行共培养过夜。对淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)设门后，通过流式细胞术评价4-1bb的表达。结果示于图42中。将自体apc在37℃下用浓度逐渐降低的25-个氨基酸的肽(对应于wtp53-y220或突变的p53-y220c序列)脉冲2小时。将来自患者4259的til片段培养物(no.6)与肽-脉冲的apc共培养。对于淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)设门后，通过流式细胞术测定4-1bb的表达。结果示于图43中。将自体抗原呈递细胞在37℃下用dmso、wtp53-y220肽rntfrhsvvvpye(seqidno:533)或突变的p53-y220c肽rntfrhsvvvpce(seqidno:534)脉冲2小时。将过量肽冲走。将来自患者4259的对p53-y220c具有特异性的til在37℃下与肽-脉冲的apc共培养过夜。对于淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)设门后，通过流式细胞术评价4-1bb的表达。结果示于图44中。将cos7细胞(每孔2.5x104个)涂铺在平底96孔板的孔上。接下来的一天，将细胞与来自患者4259的单独的hla等位基因共转染。次日，将dmso或p53-y220c肽rntfrhsvvvpce(seqidno:534)在经转染的cos7细胞上脉冲2小时。将过量肽冲走。添加来自患者4259的til片段培养物no.6(105个细胞/孔)。将共培养物在37℃下孵育过夜。对于淋巴细胞→活→cd3+(t细胞)→cd8-cd4+设门后，通过流式细胞术测定4-1bb的表达。结果示于图45中。将患者4259自体的apc在37℃下用对应于wtp53-y220或经突变的p53-y220c序列的25-个氨基酸的肽脉冲2小时。将过量肽冲走。将表达4259-f6-tcr的t细胞与肽-脉冲的apc在37℃下共培养过夜。对于淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)设门后，通过流式细胞术评价4-1bb的表达。通过小鼠tcrβ(mtcr)测量导入的tcr。结果示于表45中。表45wtp53-y220mutp53-y220c4-1bb+/mtcr+0.275.224-1bb-/mtcr-68.573.74-1bb+/mtcr-0.181.594-1bb-/mtcr+31.019.5肿瘤细胞(tc)系由从患者4259切除的异种移植的肿瘤碎片建立，然后通过免疫受损的小鼠连续传代(tc#4259)。将tc#4259与表达模拟物(无tcr)或p53-y220c-特异性tcr(4259-f6-tcr)的t细胞(105)在37℃下共培养过夜。将单独的tc#4259细胞或者将tc#4259细胞与w6/32泛-hlai类特异性阻断抗体、iva12泛-hlaii类特异性阻断抗体或经突变的p53-y220c肽rntfrhsvvvpce(seqidno:534)在37℃下孵育2小时。将抗体以5μg/ml的终浓度保存在共培养物中。将肽以10μg/ml孵育，并且孵育后洗涤过量肽。单独的培养基(无tc)及pma和离子霉素分别为阴性对照和阳性对照。对于淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)设门后，通过流式细胞术评价4-1bb的表达。结果示于图46中。从患者4259分离的tcr4259-f6-tcr的序列示于以下。从氨基末端开始，第一加下划线的区域是cdr1α(seqidno:477)，第二加下划线的区域是cdr2α(seqidno:478)，第三加下划线的区域是cdr3α(seqidno:479)，第四加下划线的区域是cdr1β(seqidno:480)，第五加下划线的区域是cdr2β(seqidno:481)，和第六加下划线的区域是cdr3β(seqidno:482)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一斜体区是α链恒定区(seqidno:23)并且第二斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:483)包含从氨基末端开始并且就在α链恒定区开始之前结束的序列。β链可变区(seqidno:484)包含在连接子之后立即开始并且就在β链恒定区开始之前结束的序列。全长α链(seqidno:485)包含从氨基末端开始并且就在连接子开始之前结束的序列。全长β链(seqidno:486)包含在连接子之后立即开始并且以羧基末端结束的序列。如下所述鉴定癌症反应性t细胞。如下所述分离tcr。tcr名称:4259-f6-tcrp53突变的识别:y220c筛选方法:p53“热点”突变通用筛选共培养以鉴定tcr:共培养4259-f6与p53-y220c肽，和分选cd4+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:81.1％(在44对中观察到36次)tcr取向:α-β表达载体:sb转座子来自患者4259的4259-f6-tcr的统计数据示于下表46中。表46参数#频率总孔96100％cdr3α4142.7％cdr3β4749.0％4259-f6-tcr对3637.5％总配对的tcr4445.8％实施例14该实施例证实了来自患者4285的tcr的分离和特异反应性。将来自患者4285的til片段(f1-f22和f24，n＝23)与用由无关、wtp53或经突变的p53序列组成的tmg电穿孔的自体apc共培养。在37℃下进行共培养过夜。使用elispot测定评价ifn-γ的分泌。结果显示于图47中。对于淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)设门后，通过流式细胞术评价4-1bb的表达。结果显示于图49中。将来自患者4285的til片段(f1-f22和f24，n＝23)与用肽媒介物(dmso)或由wtp53-r175序列或突变的p53-r175h序列组成的经纯化的(>95％，通过hplc)25个氨基酸的肽脉冲的自体apc共培养。在37℃下进行共培养过夜。使用elispot测定评价ifn-γ的分泌。结果显示于图48中。对于淋巴细胞→活细胞(pi阴性)→cd3+(t细胞)设门后，通过流式细胞术评价4-1bb的表达。结果显示于图50中。til片段f6是分选cd4+41bb+t细胞后的4285-tcr1的来源。将自体apc用来自p53-r175h序列(表45中加下划线的氨基酸取代)的重叠14个氨基酸的15个氨基酸的肽脉冲。将对p53-r175h具有特异性的来自患者4285的til(片段培养物10、6和9)与肽-脉冲的apc在37℃下共培养过夜。dmso是肽媒介物。使用elispot测定评价ifn-γ的分泌。结果显示于表47中。表47将cos7细胞(每孔2.5x104个)涂铺在平底96孔板的孔上。第二天，将细胞与来自患者4285的单独hla等位基因共转染。次日，将细胞用dmso或经突变的p53-r175h肽ykqsqhmtevvrhcphhercsdsdg(seqidno:2)在37℃下在10μg/ml下脉冲2小时。将来自患者4285的对p53-r175h具有特异性的选定til片段培养物(4285-f6、4285-f9和4285-f10)与经转染的cos7细胞在37℃下共培养过夜。对于淋巴细胞→活→cd3+(t细胞)→cd8-cd4+设门后，通过流式细胞术测定4-1bb的表达。结果显示于图51中。将自体apc用对应于wt或突变的p53-r175h序列的逐渐降低浓度的25-个或15-个氨基酸的肽在37℃下脉冲2小时。将来自患者4285的4285-tcr1转座的t细胞与肽-脉冲的apc在37℃下共培养过夜。对于淋巴细胞→活→cd3+(t细胞)→cd8-cd4+设门后，通过流式细胞术测定4-1bb的表达。结果显示于图52中。从患者4285分离的4285-tcr1的序列示于下文。从氨基末端开始，第一加下划线区是cdr1α(seqidno:487)，第二加下划线区是cdr2α(seqidno:488)，第三加下划线区是cdr3α(seqidno:489)，第四加下划线区是cdr1β(seqidno:490)，第五加下划线区是cdr2β(seqidno:491)，和第六加下划线区是cdr3β(seqidno:492)。粗体区是连接子(seqidno:26)。从氨基末端开始，第一加斜体区是α链恒定区(seqidno:23)并且第二加斜体区是β链恒定区(seqidno:25)。α链可变区(seqidno:493)包含从氨基末端开始并且就在α链恒定区开始之前结束的序列。β链可变区(seqidno:494)包含在连接子之后立即开始并且就在β链恒定区开始之前结束的序列。全长α链(seqidno:495)包含从氨基末端开始并且就在连接子开始之前结束的序列。全长β链(seqidno:496)包含在连接子之后立即开始并且以羧基末端结束的序列。癌症反应性t细胞如下所述鉴定。tcr如下所述分离。tcr名称:4285-tcr1p53突变的识别:r175h筛选方法:p53“热点”突变通用筛选共培养以鉴定tcr:共培养4285-f6与p53-r175h肽和分选cd4+41bb+t细胞鉴定tcr的方法:单细胞rt-pcr所有配对的tcr中的tcr的丰度:81.8％(在44对中观察到36次)tcr取向:α-β表达载体:sb转座子患者4285的4285-tcr1的统计数据示于下表48中。表48参数#频率总孔96100％cdr3α3739.8％cdr3β3940.6％4285-tcr1对3637.5％总配对的tcr4445.8％实施例15该实施例验证了来自实施例2的患者4196的三种抗突变p53tcr的特异反应性。不同靶细胞系的hla-a*0201和p53r175h的表达如表49所示。表49将表49中的靶细胞与用实施例2中的一种tcr转导的细胞共培养。使用模拟转导细胞(无tcr)作为对照效应细胞。测定ifn-γ分泌(pg/ml)(表50)和4-1bb表达(％4-1bb(mtcrβ+的))(表51)。表50表51实施例16该实施例证实了实施例1-15的tcr的反应性的概述。将表52的tcr分离，在t细胞中表达并针对相关抗原进行了测试。结果概述显示于表52中。表52本文引用的所有参考文献，包括出版物、专利申请和专利，均通过引用并入本文，其程度如同每篇参考文献均被单独且具体地指示为通过引用并入本文并在本文中整体阐述。在描述本发明的上下文中(尤其是在所附权利要求的上下文中)术语“一个”和“一种”和“该”和“至少一种”以及类似指代物的使用应被解释为涵盖了单数和复数两种，除非本文另有说明或与上下文明显矛盾。术语“至少一种”之后是一项或多项的列表(例如，“a和b中的至少一种”)的使用应理解为意指选自所列项(a或b)的一项或所列项(a和b)的两种或多种的任何组合，除非本文另有说明或与上下文明显矛盾。除非另有说明，否则术语“包含”、“具有”、“包括”和“含有”应被解释为开放式术语(即，意指“包括但不限于”)。除非在本文中另外指出，否则本文中数值范围的叙述仅旨在用作分别指代落入该范围内的每个单独值的简写方法，并且每个单独值都被并入说明书中，就好像它在本文中被单独叙述一样。除非本文另外指出或另外与上下文明显矛盾，否则本文描述的所有方法可以以任何适合的顺序执行。除非另外要求保护，否则本文提供的任何和所有示例或示例性语言(如，“诸如”)的使用仅旨在更好地阐明本发明，并且不对本发明的范围构成限制。说明书中的任何语言都不应解释为指示任何未要求保护的要素对于实施本发明必不可少。本文描述了本发明的优选实施方案，包括发明人已知的用于实施本发明的最佳模式。在阅读前述说明书之后，那些优选实施方案的变型对于本领域普通技术人员而言将变得显而易见。本发明人期望熟练的技术人员适当地采用这样的变型，并且本发明人有意以不同于本文具体描述的方式来实践本发明。因此，本发明包括适用法律所允许的所附权利要求书中记载的主题的所有修改和等同物。此外，除非本文另外指出或另外与上下文明显矛盾，否则本发明涵盖上述要素在其所有可能的变型中的任何组合。序列表<110>theunitedstatesofamerica,asrepresentedbythesecretary,departmentofhealthandhumanservices<120>识别突变的p53的t细胞受体<130>739980<150>us62/565,383<151>2017-09-29<160>602<170>patentinversion3.5<210>1<211>393<212>prt<213>人<400>1metglugluproglnseraspproservalgluproproleusergln151015gluthrpheseraspleutrplysleuleuprogluasnasnvalleu202530serproleuproserglnalametaspaspleumetleuserproasp354045aspilegluglntrpphethrgluaspproglyproaspglualapro505560argmetproglualaalaproprovalalaproalaproalaalapro65707580thrproalaalaproalaproalaprosertrpproleuserserser859095valproserglnlysthrtyrglnglysertyrglypheargleugly100105110pheleuhisserglythralalysservalthrcysthrtyrserpro115120125alal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