Schrader J W, Edelman G M
Rockefeller University, New York 10021.
J Exp Med. 1977 Mar 1;145(3):523-39. doi: 10.1084/jem.145.3.523.
Cytotoxic T cells specific for Sendai virus were generated by culturing murine spleen cells in vitro together with UV-inactivated Sendai virus. In vivo immunization of donor mice with UV-inactivated Sendai virus resulted in an in vitro secondary response of increased magnitude. Cytotoxic activity was demonstrated in a short-term 51Cr-release assay, using syngeneic tumor cells which had been coated with inactivated Sendai virus by incubation at 4 degrees C for 30 min. The lysis of Sendai virus-coated target cells was restricted by the H-2 haplotype of the target cells, suggesting that the H-2 genes of the target cell contributed to the specificity of the lysis. Kinetic experiments showed that susceptibility to lysis by cytotoxic T cells specific for Sendai virus appeared within 30 min after coating target cells with inactivated virus. Furthermore, there was no detectable synthesis of new proteins in cells treated with UV-inactivated Sendai virus. For these reasons, we suggest that neither viral replication nor the synthesis of new proteins are necessary for the production of the antigen recognized by cytotoxic cells specific for Sendai virus. We infer that the virus-specific component on the target cells is probably a preformed virion antigen adsorbed onto or integrated into the cell membrane. These results imply that, if the cytotoxic T cell recognizes a single antigenic determinant specified both by viral and H-2 genes, this determinant is formed by the physical association of H-2 and Sendai virus antigens rather than by their alteration during the processes of synthesis.
通过将小鼠脾细胞与紫外线灭活的仙台病毒在体外共同培养,产生了对仙台病毒具有特异性的细胞毒性T细胞。用紫外线灭活的仙台病毒对供体小鼠进行体内免疫,可导致体外二级反应的幅度增加。在短期的51Cr释放试验中证明了细胞毒性活性,该试验使用了通过在4℃孵育30分钟而被灭活的仙台病毒包被的同基因肿瘤细胞。仙台病毒包被的靶细胞的裂解受靶细胞的H-2单倍型限制,这表明靶细胞的H-2基因促成了裂解的特异性。动力学实验表明,在用灭活病毒包被靶细胞后30分钟内,对仙台病毒特异性细胞毒性T细胞的裂解敏感性就会出现。此外,在用紫外线灭活的仙台病毒处理的细胞中未检测到新蛋白质的合成。基于这些原因,我们认为病毒复制和新蛋白质的合成对于产生被仙台病毒特异性细胞毒性细胞识别的抗原都不是必需的。我们推断靶细胞上的病毒特异性成分可能是吸附在细胞膜上或整合到细胞膜中的预先形成的病毒粒子抗原。这些结果意味着,如果细胞毒性T细胞识别由病毒和H-2基因共同指定的单一抗原决定簇,那么这个决定簇是由H-2和仙台病毒抗原的物理结合形成的,而不是在合成过程中由它们的改变形成的。