Tanaka Y, Anderson R W, Maloy W L, Tevethia S S
Department of Microbiology and Immunology, Penn State University, College of Medicine, Hershey 17033.
Virology. 1989 Jul;171(1):205-13. doi: 10.1016/0042-6822(89)90527-8.
SV40 tumor (T) antigen possesses four distinct antigenic determinants, sites I, II, III, and IV, recognized by SV40-specific H-2b-restricted cytotoxic T-lymphocytes (CTL) clones. SV40-transformed C57BL/6 (B6) mouse kidney cells, designated K-3, 1, 4, K-1, 4, and K-4, 1, have been isolated by immunological selection with SV40 T antigen site-specific CTL clones in vitro. The cells have lost the expression of all four antigenic sites and cannot be lysed by the CTL clones specific for antigenic sites I, II, III, and IV. To search for additional SV40-specific antigenic sites on SV40 T antigen, B6 mice were immunized with K-3,1,4 cells and stimulated spleen cells with K-3,1,4 cells in vitro. Repeated stimulation of the spleen cell culture with gamma-irradiated K-3,1,4 cells in the presence of IL-2 was necessary to generate CTL activity against K-3,1,4 cells. A new group of H-2Db-restricted CTL clones designated as Y-5 was isolated which were cytotoxic to K-3,1,4 cells. The antigenic site recognized by CTL clone Y-5, site V, was localized in the carboxy terminal half of the SV40 T antigen. By the use of a synthetic peptide corresponding to SV40 T antigen in the carboxy region, the antigenic site V was localized between amino acids 489 and 503.
SV40肿瘤（T）抗原具有四个不同的抗原决定簇，即位点I、II、III和IV，可被SV40特异性的H-2b限制性细胞毒性T淋巴细胞（CTL）克隆识别。通过在体外使用SV40 T抗原位点特异性CTL克隆进行免疫选择，已分离出SV40转化的C57BL/6（B6）小鼠肾细胞，命名为K-3,1,4、K-1,4和K-4,1。这些细胞已失去所有四个抗原位点的表达，并且不能被针对抗原位点I、II、III和IV的CTL克隆裂解。为了在SV40 T抗原上寻找其他SV40特异性抗原位点，用K-3,1,4细胞免疫B6小鼠，并在体外使用K-3,1,4细胞刺激脾细胞。为了产生针对K-3,1,4细胞的CTL活性，需要在IL-2存在下用γ射线照射的K-3,1,4细胞反复刺激脾细胞培养物。分离出一组新的H-2Db限制性CTL克隆，命名为Y-5，它们对K-3,1,4细胞具有细胞毒性。CTL克隆Y-5识别的抗原位点，即位点V，位于SV40 T抗原的羧基末端一半区域。通过使用与SV40 T抗原羧基区域相对应的合成肽，抗原位点V定位在氨基酸489和503之间。
