Schirmbeck R, Deppert W, Kury E, Reimann J
Institute of Microbiology, Universität Ulm, Germany.
Cell Immunol. 1993 Jul;149(2):444-9. doi: 10.1006/cimm.1993.1169.
Injection of the 708-amino-acid (aa) viral protein "large tumor antigen" (T-Ag) of simian virus 40 (SV40) or its N-terminal 272-aa fragment into C57BL/6 (B6) mice (H-2b) primed CD8+ cytotoxic T lymphocytes (CTL) in vivo. Surprisingly, injection of this nonstructural viral protein (or its N-terminal fragment) in soluble form (without adjuvants) was as efficient in priming CD8+ CTL in vivo as the infection of B6 mice with the virus SV40. CTL activated in vivo by immunization with T-Ag proteins or SV40 infection specifically lysed syngeneic RBL5 cells transfected with a T-Ag-encoding vector; these RBL5/M7 transfectants efficiently presented N- and C-terminal T-Ag epitopes in association with H-2 class I restriction elements. N- and C-terminal T-Ag epitopes were recognized by CTL primed in vivo by immunization with the complete T-Ag protein or by infection with SV40, and (as expected) only N-terminal T-Ag epitopes were recognized by CTL primed in vivo by the soluble N-terminal T-Ag fragment. In CD8+ CTL populations primed in vivo by immunization with the complete T-Ag protein or by SV40 infection and restimulated in vitro with RBL5/M7 transfectants in a mixed tumor cell-lymphocyte culture (MTLC), CTL with specificity for C-terminal T-Ag epitopes were selectively expanded in vitro for months. Hence, the in vitro expansion of CTL population with heterogenous recognition specificities can dramatically distort the picture of its specific recognition repertoire primed in vivo.
将猿猴病毒40（SV40）的708个氨基酸（aa）的病毒蛋白“大肿瘤抗原”（T-Ag）或其N端272个氨基酸的片段注射到C57BL/6（B6）小鼠（H-2b）体内，可在体内启动CD8+细胞毒性T淋巴细胞（CTL）。令人惊讶的是，以可溶性形式（无佐剂）注射这种非结构病毒蛋白（或其N端片段）在体内启动CD8+CTL的效率与用SV40病毒感染B6小鼠相同。通过用T-Ag蛋白免疫或SV40感染在体内激活的CTL特异性裂解用编码T-Ag的载体转染的同基因RBL5细胞；这些RBL5/M7转染子与H-2 I类限制元件相关联，有效地呈递N端和C端T-Ag表位。N端和C端T-Ag表位可被用完整T-Ag蛋白免疫或用SV40感染在体内启动的CTL识别，并且（如预期的那样）只有N端T-Ag表位可被用可溶性N端T-Ag片段在体内启动的CTL识别。在用完整T-Ag蛋白免疫或SV40感染在体内启动并在混合肿瘤细胞-淋巴细胞培养（MTLC）中用RBL5/M7转染子在体外再刺激的CD8+CTL群体中，对C端T-Ag表位具有特异性的CTL在体外可选择性扩增数月。因此，具有异源识别特异性的CTL群体的体外扩增可极大地扭曲其在体内启动的特异性识别库的情况。
