Frequency analysis of simian virus 40-specific cytotoxic T lymphocyte precursors in the high responder C57BL/6 mouse strain.
Frequency analysis of simian virus 40-specific cytotoxic T lymphocyte precursors in the high responder C57BL/6 mouse strain.
复制标题
高反应 C57BL/6 小鼠品系中猿猴病毒 40 特异性细胞毒性 T 淋巴细胞前体的频率分析。
DOI：
10.1099/0022-1317-69-10-2493
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发表时间：
1988
期刊：
影响因子：
--
通讯作者：
Tevethia,SS
中科院分区：
文献类型：
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作者：
Jennings,SR;Fresa,KL;Lippe,PA;Milici,JE;Tevethia,SS
本实验室的研究表明，从高应答C57 BL/6（B6; H-2b）小鼠脾脏建立的长期猴病毒40（SV 40）特异性溶细胞性T淋巴细胞（CTL）培养物显示出对H-2Db限制性CTL克隆的选择偏好。在这项研究中，我们调查了这种选择的基础。使用来自B6小鼠的腘淋巴结和脾脏的应答细胞建立有限稀释培养物，所述B6小鼠分别在后足垫中皮下免疫或通过腹膜内途径免疫，具有表达全长（1至708个氨基酸残基）SV 40大T抗原的同基因SV 40转化细胞。CTL前体（CTL precursor，CTLp）在SV 40转化的刺激细胞和白细胞介素2存在下能在体外扩增，并对表达全长T抗原的H-2b细胞表现出裂解活性，在腘淋巴结中的相对频率为1/1900 ~ 1/15000，在脾脏中的相对频率为1/8000 ~ 1/55000。在这两个实验系统中，H-2Kb限制的CTLp的出现频率明显高于H-2Db限制的CTLp。此外，在T抗原的氨基末端或羧基末端的一半内产生的CTLp识别决定簇的数量大致相等。用抗Lyt 2单克隆抗体对SV 40特异性CTL的相对亲和力进行了测定，结果表明，H-2Db限制性CTL与靶细胞的相互作用亲和力大于H-2Kb限制性CTL。这些数据表明，H-2Db限制性CTL克隆的优势分离长期体外培养可能是这个群体作为一个整体的相对亲和力的功能，而不是由于该亚群在体内对SV 40 T抗原的免疫优势。
Studies in this laboratory have shown that long term simian virus 40 (SV40)-specific cytolytic T lymphocyte (CTL) cultures established from the spleens of high responder C57BL/6 (B6; H-2b) mice exhibit a preference for the selection of H-2Db-restricted CTL clones. In this study, we have investigated the basis for this selection. Limiting dilution cultures were established using responder cells from the popliteal lymph nodes and the spleens of B6 mice immunized subcutaneously in the hind footpads or via the intraperitoneal route, respectively, with syngeneic SV40-transformed cells expressing a full length (1 to 708 amino acid residues) SV40 large T antigen. The relative frequency of CTL precursors (CTLp) able to expandin vitroin the presence of SV40-transformed stimulator cells and interleukin 2 and exhibit lytic activity against H-2bcells expressing full length T antigen ranged from 1/1900 to 1/15000 in the popliteal lymph node and from 1/8000 to 1/55000 in the spleen. In these two experimental systems, CTLp restricted to H-2Kbwere apparently present at higher frequency than H-2Db-restricted CTLp. Furthermore, CTLp recognizing determinants within the amino-terminal or carboxy-terminal halves of T antigen were generated in approximately equal numbers. The relative affinity of SV40-specific CTL, assessed by inhibition with anti-Lyt 2 monoclonal antibody, indicated that CTL restricted to H-2Dbinteracted with their target with greater affinity than CTL restricted to H-2Kb. These data suggest that the predominance of isolation of H-2Db-restricted CTL clones from long termin vitrocultures may be a function of the relative affinity of this population as a whole, rather than due to the immunodominance of this subpopulation during thein vivoresponse to SV40 T antigen.