Haskins K, Hannum C, White J, Roehm N, Kubo R, Kappler J, Marrack P
J Exp Med. 1984 Aug 1;160(2):452-71. doi: 10.1084/jem.160.2.452.
We have prepared a monoclonal antibody, KJ16-133, from the cells of a rat immunized with the purified receptor for antigen plus I-A of a BALB/c T cell hybridoma, DO-11.10. Unlike most other monoclonal anti-receptor antibodies that have been described before, KJ16-133 is not clone specific. It reacts with approximately 20% of the receptors on T cells of normal BALB/c mice. It also reacts with about the same percentage of antigen-specific, major histocompatibility complex (MHC)-restricted or allogeneic I-region specific T cell hybridomas. Reaction of KJ16-133 with a given T cell hybridoma does not seem to depend on the antigen specificity or MHC-restricting element of the T cell in question. The determinant recognized by KJ16-133 has some unexpected properties. It is absent in several strains of mice including SJL/J and SJA/20, but present on the T cells of most other commonly used strains. The determinant recognized therefore does not map to Igh. Our experiments suggest that a clone-specific "antiidiotypic" antibody and KJ16-133 recognize determinants on different parts of the receptor. For example, the binding of a clone-specific antibody to target T cells is relatively temperature insensitive, whereas KJ16-133 binds well to cells at 37 degrees C but poorly to cells at 4 degrees C. The determinant recognized by a clone-specific antibody is sensitive to reduction and alkylation of the receptor, whereas KJ16-133 reactivity is not. Finally, binding of KJ16-133 at saturating concentrations to target T cells does not block the binding of a clone-specific antibody. Similarly, binding of a clone-specific antibody only marginally inhibits binding of KJ16-133. Taken together, these results suggest that KJ16-133 is directed against an allelic determinant on T cells that may be close to the membrane, and not in the receptor binding site for antigen plus MHC. The antibody may recognize an allele of a constant region isotype, or an allele of a J region.
我们用纯化的抗原受体加BALB/c T细胞杂交瘤DO-11.10的I-A免疫大鼠的细胞，制备了一种单克隆抗体KJ16-133。与之前描述的大多数其他单克隆抗受体抗体不同，KJ16-133不是克隆特异性的。它能与正常BALB/c小鼠T细胞上约20%的受体发生反应。它也能与大约相同比例的抗原特异性、主要组织相容性复合体（MHC）限制的或同种异体I区特异性T细胞杂交瘤发生反应。KJ16-133与特定T细胞杂交瘤的反应似乎不取决于所讨论的T细胞的抗原特异性或MHC限制元件。KJ16-133识别的决定簇具有一些意想不到的特性。它在包括SJL/J和SJA/20在内的几种小鼠品系中不存在，但在大多数其他常用品系的T细胞上存在。因此，所识别的决定簇并不定位于Igh。我们的实验表明，一种克隆特异性的“抗独特型”抗体和KJ16-133识别受体不同部位的决定簇。例如，克隆特异性抗体与靶T细胞的结合对温度相对不敏感，而KJ16-133在37℃时能很好地结合细胞，但在4℃时与细胞的结合很差。克隆特异性抗体识别的决定簇对受体的还原和烷基化敏感，而KJ16-133的反应性则不敏感。最后，饱和浓度的KJ16-133与靶T细胞的结合并不阻断克隆特异性抗体的结合。同样，克隆特异性抗体的结合仅略微抑制KJ16-133的结合。综上所述，这些结果表明，KJ16-133针对的是T细胞上可能靠近细胞膜的一个等位基因决定簇，而不是抗原加MHC的受体结合位点。该抗体可能识别恒定区同种型的一个等位基因，或J区的一个等位基因。
