Matsubayashi Y, Hirama T, Morioka A, Iwashiro M, Masuda T, Uchino H, Takeshita S, Yamagishi H, Udono H, Mieno M
Institute for Immunology, Faculty of Medicine, Kyoto University, Japan.
Eur J Immunol. 1990 Sep;20(9):2095-103. doi: 10.1002/eji.1830200931.
Cytotoxic T lymphocyte (CTL) clones against a syngeneic Friend virus-induced erythroleukemia (FBL-3) were generated in C57BL/6 (B6) mice. A monoclonal antibody (mAb, N9-127) was then raised from spleen cells of a B6 mouse immunized syngenically against one of these CTL clones. This mAb detected the epitope (127Ep) of the T cell antigen receptor (TcR) on the immunizing CTL clone in tests of immunoprecipitation, specific blocking and proliferation, and induction of TcR-mediated nonspecific lysis of the clone. In addition, more than 10% of the FBL-3-specific CTL clones isolated independently from B6 mice were 127Ep+. Further investigations revealed that up to 30% of B6 anti-FBL-3 T cell blasts from mixed lymphocyte tumor cell cultures were positive for this epitope, and that its expression was confined to CD8+ T cells. This epitope was not detected in naive lymphoid cells from the spleen, lymph nodes or thymus or in T cell clones specific for tumors other than FBL-3. The FBL-3-specific CTL clones were next grouped into 127Ep+ and 127Ep- clones. Sequence analyses of the CTL clone used for immunization showed the rearrangements of V alpha 1J alpha 112-2 and V beta 10D beta 2.1J beta 2.7. Southern blot analysis of all the 127Ep+ CTL clones examined showed the same DNA rearrangement bands of both the TcR alpha and beta genes. These findings suggested that mAb N9-127 recognized the shared determinant of the TcR molecule which was expressed by the dominant CTL population in the response to FBL-3.
在C57BL/6(B6)小鼠中产生了针对同基因Friend病毒诱导的红白血病(FBL-3)的细胞毒性T淋巴细胞(CTL)克隆。然后从同基因免疫了其中一个CTL克隆的B6小鼠的脾细胞中制备了一种单克隆抗体(mAb,N9-127)。在免疫沉淀、特异性阻断和增殖以及诱导该克隆的TcR介导的非特异性裂解试验中,该mAb检测到了免疫化CTL克隆上T细胞抗原受体(TcR)的表位(127Ep)。此外,从B6小鼠中独立分离的超过10%的FBL-3特异性CTL克隆为127Ep阳性。进一步研究表明,在混合淋巴细胞肿瘤细胞培养物中,高达30%的B6抗FBL-3 T细胞母细胞对该表位呈阳性,并且其表达仅限于CD8+ T细胞。在来自脾脏、淋巴结或胸腺的未活化淋巴细胞或针对FBL-3以外肿瘤的T细胞克隆中未检测到该表位。接下来,将FBL-3特异性CTL克隆分为127Ep阳性和127Ep阴性克隆。用于免疫的CTL克隆的序列分析显示Vα1Jα112-2和Vβ10Dβ2.1Jβ2.7发生了重排。对所有检测的127Ep阳性CTL克隆进行的Southern印迹分析显示,TcRα和β基因具有相同的DNA重排条带。这些发现表明,mAb N9-127识别了TcR分子的共享决定簇,该决定簇由对FBL-3反应中的优势CTL群体表达。