A MOUSE MUTANT P53 PRODUCT RECOGNIZED BY CD4+ AND CD8+ T-CELLS
A MOUSE MUTANT P53 PRODUCT RECOGNIZED BY CD4+ AND CD8+ T-CELLS
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DOI：
10.1073/pnas.91.8.3171
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发表时间：
1994-04-12
影响因子：
11.1
通讯作者：
OLD, LJ
中科院分区：
文献类型：
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作者：
NOGUCHI, Y;CHEN, YT;OLD, LJ
分析了T细胞对BALB/c衍生的MethA肉瘤的突变和正常p53产物的应答。已知方法A p53在密码子132、168和234中具有三个错义点突变，并且构建了在三个突变位点处含有野生型或突变序列的24种肽。用p53肽免疫BALB/c或（BALB/c × C57 BL/6）F1小鼠，取脾细胞，用相应的肽体外致敏。因为甲氧苯甲酸对细胞毒性T细胞具有抗性，所以选择表达低水平p53但缺乏甲氧苯甲酸p53突变的敏感P1-HTR细胞系作为靶标，用p53肽脉冲标记或用含有来自甲氧苯甲酸p53的编码序列的质粒转染。一种肽，含有密码子234突变的九聚体（234 CM），诱导CD 8+细胞毒性T细胞以H-2K（d）限制性方式裂解234 CM脉冲的P1-HTR细胞。用相应的野生型肽脉冲的P1-HTR细胞仅被234 CM反应性细胞毒性T细胞微弱地裂解。用其他野生型或突变型p53肽脉冲的P1-HTR细胞不被234 CM反应性细胞毒性T细胞裂解，这些肽，包括234 CW（234 CM的野生型对应物）也不能引起细胞毒性细胞。用编码234 CM序列的质粒转染并表达高水平p53的P1-HTR细胞被234 CM反应性细胞毒性T细胞微弱裂解。然而，裂解的转染子之一显着增加了干扰素γ预处理。用234 CM和234 CW免疫引起CD 4 + T细胞的增殖反应，但不与其他p53相关肽。234 CM诱导的CD 4 + T细胞对234-区域肽的特异性比234 CM反应性细胞毒性T细胞的反应性更广泛。在不完全弗氏佐剂中用234 CM免疫的小鼠显示出对甲硫氨酸攻击的增强的抗性。
The T-cell response to mutated and normal p53 products of BALB/c-derived Meth A sarcoma was analyzed. Meth A p53 is known to have three missense point mutations in codons 132, 168, and 234, and 24 peptides containing wild-type or mutated sequences at the three mutation sites were constructed. Spleen cells from BALB/c or (BALB/c x C57BL/6)F1 mice immunized with p53 peptides were sensitized in vitro with the corresponding peptides. Because Meth A is resistant to cytotoxic T cells, the sensitive P1-HTR cell line, which expresses a low level of p53 lacking the Meth A p53 mutations, was chosen as a target, either pulse-labeled with p53 peptides or transfected with plasmids containing coding sequences from Meth A p53. One peptide, a nonamer containing the codon 234 mutation (234CM), induced CD8+ cytotoxic T cells that lysed 234CM-pulsed P1-HTR cells in an H-2K(d)-restricted fashion. P1-HTR cells pulsed with the corresponding wild-type peptide were only weakly lysed by 234CM-reactive cytotoxic T cells. P1-HTR cells pulsed with other wild-type or mutated p53 peptides were not lysed by 234CM-reactive cytotoxic T cells, nor could these peptides, including 234CW (the wild-type counterpart to 234CM), elicit cytotoxic cells. P1-HTR cells transfected with plasmids coding for the 234CM sequence and expressing high p53 levels were weakly lysed by 234CM-reactive cytotoxic T cells. However, lysis of one of the transfectants was significantly increased by pretreatment with interferon gamma. A proliferative response of CD4+ T cells was elicited by immunization with 234CM and 234CW, but not with other p53-related peptides. The specificity of 234CM-induced CD4+ T cells for 234-region peptides was broader than the reactivity of 234CM-reactive cytotoxic T cells. Mice immunized with 234CM in incomplete Freund's adjuvant showed heightened resistance to Meth A challenge.