Kaufman J, Ferrone S, Flajnik M, Kilb M, Völk H, Parisot R
Basel Institute for Immunology, Switzerland.
J Immunol. 1990 Mar 15;144(6):2273-80.
mAb to human and mouse MHC molecules were tested for binding to blood or spleen cells of various nonmammalian vertebrates by immunofluorescence and flow cytometry. Those that bound were used to immunoprecipitate cross-reactive molecules from biosynthetically or cell surface-labeled spleen or blood cells. In addition, mAb to human MHC molecules were screened by Western blots. As expected from the results with xenoantisera, there were few mAb that cross-reacted, and many of these cross-reactions were not specific for MHC-like molecules. Less than 10% of the mAb tested bound to the cells of any particular species, with very few positive for more than one species. Of those mAb that bound cells, many failed to precipitate any radioactive bands, and most bands precipitated were not recognizable as MHC-like molecules. Five mAb reacted with Xenopus class II, one of which also immunoprecipitated axolotl class II. Another of these reacted with a candidate for class II in the lamprey, but this molecule had features unlike those expected for mammalian class II molecules. Four other mAb reacted with candidate molecules. in the trout and shark. None of the mouse alloantibodies immunoprecipitated nonmammalian vertebrate MHC-like molecules. In contrast to the results with most xenoantisera, the mAb cross-reacting with amphibian class II molecules recognized a number of different linear epitopes on the surface of the polymorphic non-Ig beta 1 domain of class II molecules. Few mAb recognized bands in Western blots of nonmammalian vertebrate cells and the candidate molecules from fish had features different from known mammalian MHC molecules.
通过免疫荧光和流式细胞术,检测针对人类和小鼠MHC分子的单克隆抗体与各种非哺乳动物脊椎动物的血液或脾细胞的结合情况。那些能够结合的抗体被用于从生物合成或细胞表面标记的脾细胞或血细胞中免疫沉淀交叉反应分子。此外,通过蛋白质免疫印迹法筛选针对人类MHC分子的单克隆抗体。正如异种抗血清的结果所预期的那样,很少有单克隆抗体发生交叉反应,而且其中许多交叉反应并非针对MHC类分子具有特异性。所检测的单克隆抗体中,不到10%与任何特定物种的细胞结合,对不止一个物种呈阳性反应的非常少。在那些能够结合细胞的单克隆抗体中,许多未能沉淀出任何放射性条带,而且沉淀出的大多数条带无法识别为MHC类分子。五种单克隆抗体与非洲爪蟾的II类分子发生反应,其中一种还免疫沉淀出美西螈的II类分子。其中另一种与七鳃鳗中一种II类候选分子发生反应,但该分子具有与哺乳动物II类分子预期特征不同的特点。另外四种单克隆抗体与鳟鱼和鲨鱼中的候选分子发生反应。没有一种小鼠同种异体抗体免疫沉淀出非哺乳动物脊椎动物的MHC类分子。与大多数异种抗血清的结果相反,与两栖动物II类分子发生交叉反应的单克隆抗体识别出II类分子多态性非Igβ1结构域表面的许多不同线性表位。很少有单克隆抗体在非哺乳动物脊椎动物细胞的蛋白质免疫印迹中识别出条带,而且来自鱼类的候选分子具有与已知哺乳动物MHC分子不同的特征。