Marrack P, Shimonkevitz R, Hannum C, Haskins K, Kappler J
J Exp Med. 1983 Nov 1;158(5):1635-46. doi: 10.1084/jem.158.5.1635.
In order to prove that a monoclonal antibody, KJ1-26.1, reacted with the idiotypic portion of the receptor for antigen plus major histocompatibility complex product (Ag/MHC) on the T cell hybridoma D0-11.10, 397 independent T cell hybridomas prepared from BALB/c T cells primed with ovalbumin were screened by ELISA for reactivity with the antibody. Of these T cell hybridomas one, 7D0-286, and a subclone of this hybridoma, 7D0-286.2, reacted strongly and consistently with KJ1-26.1. Further analysis revealed that KJ1-26.1 blocked the reaction of 7D0-286 with Ag/MHC, and that 7D0-286 had the same fine specificity both for Ag and MHC as D0-11.10. None of 207 other BALB/c T cell hybridomas specific for ovalbumin shared these fine specificity patterns. Thus, reaction with KJ1-26.1 defined the specificity of a T cell hybridoma both for Ag and MHC, suggesting that KJ1-26.1 reacted with the receptors for Ag/MHC on the T cells in question. KJ1-26.1 precipitated an 85-kdalton molecule from 7D0-286.2 that on reduction ran as a diffuse spot of approximately 43 kdaltons. A similar molecule was precipitated from D0-11.10.
为了证明单克隆抗体KJ1-26.1与T细胞杂交瘤D0-11.10上抗原加主要组织相容性复合体产物(Ag/MHC)受体的独特型部分发生反应,通过酶联免疫吸附测定(ELISA)筛选了397个由用卵清蛋白致敏的BALB/c T细胞制备的独立T细胞杂交瘤与该抗体的反应性。在这些T细胞杂交瘤中,一个名为7D0-286的杂交瘤及其亚克隆7D0-286.2与KJ1-26.1发生强烈且一致的反应。进一步分析表明,KJ1-26.1阻断了7D0-286与Ag/MHC的反应,并且7D0-286对Ag和MHC的精细特异性与D0-11.10相同。另外207个对卵清蛋白特异的BALB/c T细胞杂交瘤均未共享这些精细特异性模式。因此,与KJ1-26.1的反应确定了一个T细胞杂交瘤对Ag和MHC的特异性,这表明KJ1-26.1与所研究的T细胞上的Ag/MHC受体发生了反应。KJ1-26.1从7D0-286.2沉淀出一个85千道尔顿的分子,该分子在还原后呈现为一条约43千道尔顿的弥散条带。从D0-11.10也沉淀出了类似的分子。