Moore M W, Carbone F R, Bevan M J
Department of Immunology, Research Institute of Scripps Clinic, La Jolla, California 92037.
Cell. 1988 Sep 9;54(6):777-85. doi: 10.1016/s0092-8674(88)91043-4.
In order to investigate how peptides associate with class I major histocompatibility complex (MHC) glycoproteins intracellularly, we generated cytotoxic T lymphocytes (CTL) specific for a readily available soluble protein in association with class I. C57BL/6 (H-2b) mice immunized against a syngeneic tumor cell transfected with chicken ovalbumin (OVA) cDNA gave rise to H-2Kb-restricted CTL specific for the OVA258-276 peptide. This synthetic peptide and CNBr fragments of OVA (242-285 and 242-273) were able to target H-2b cells for lysis by the CTL in a 3 hr assay. Cells incubated with native OVA for up to 24 hr did not become sensitized for recognition and lysis. However, when OVA was introduced directly into the cytoplasm of cells by the osmotic lysis of pinosomes, the Kb restricted determinant formed readily.
为了研究肽在细胞内如何与I类主要组织相容性复合体（MHC）糖蛋白结合，我们制备了对一种易于获得的与I类相关的可溶性蛋白具有特异性的细胞毒性T淋巴细胞（CTL）。用转染了鸡卵清蛋白（OVA）cDNA的同基因肿瘤细胞免疫的C57BL/6（H-2b）小鼠产生了对OVA258-276肽具有特异性的H-2Kb限制性CTL。在3小时的实验中，这种合成肽以及OVA的溴化氰片段（242-285和242-273）能够使H-2b细胞被CTL裂解。与天然OVA孵育长达24小时的细胞不会变得对识别和裂解敏感。然而，当通过胞饮体的渗透裂解将OVA直接引入细胞的细胞质中时，Kb限制性决定簇很容易形成。
