Burger R, Reske K, Mauer U, von Steldern D, Husmann M
J Immunol. 1983 Sep;131(3):1350-5.
Monoclonal antibodies (Ab) were produced that specifically recognized guinea pig T cells. FACS analysis revealed that Ab 188 bound to the majority of peripheral T lymphocytes of strain 2 and strain 13 guinea pigs and to a minor population of thymocytes. It failed to react with the Ia-bearing guinea pig B cell leukemia line EN-L2C, with macrophages, bone marrow cells, erythrocytes, or thrombocytes. Treatment of T cells with Ab 188 and complement prevented T cell activation. Culturing primed T cells with antigen- or mitogen-pulsed syngeneic or with allogeneic macrophages in the continuous presence of Ab 188 produced a marked, dose-dependent inhibition of T cell proliferation. The antigen defined by Ab 188 was therefore designated guinea pig T lymphocyte function-associated antigen-1, gp TFA-1. The magnitude of inhibition by Ab 188 varied between 65 and 85% whereas three other antibodies to guinea pig T cells had no inhibitory effect on T cell proliferation. Time course experiments revealed that gp TFA-1 is critically involved in an early phase of T cell activation. Maximal inhibition was achieved only if the antibody was present from the beginning of the cell culture; the addition of antibody after 24 hr of culture no longer had an inhibitory effect. Ab 188 did not induce T cell mitogenesis. Two-dimensional analysis (one-dimensional, IEF; two-dimensional, SDS-PAGE) of immunoprecipitates obtained from NP40 lysates of [35S]methionine-labeled T cell blasts indicated that a molecule was specifically precipitated that consisted of two noncovalently associated polypeptide chains with apparent m.w. of 43,000 and 38,000. Both subunits displayed extensive charge heterogeneity focusing at an average isoelectric point of 5.0 and 6.5, respectively. The gp TFA-1 molecule exhibits striking similarities in its functional and structural properties to recently described clonotypically expressed T cell glycoproteins, which were shown to be involved in antigen recognition by T cells in the murine and human systems.
制备出了特异性识别豚鼠T细胞的单克隆抗体(Ab)。荧光激活细胞分选术(FACS)分析显示,抗体188与2系和13系豚鼠的大多数外周T淋巴细胞以及少数胸腺细胞结合。它不与携带Ia的豚鼠B细胞白血病系EN-L2C、巨噬细胞、骨髓细胞、红细胞或血小板发生反应。用抗体188和补体处理T细胞可阻止T细胞活化。在抗体188持续存在的情况下,将致敏T细胞与抗原或丝裂原刺激的同基因或异基因巨噬细胞一起培养,可显著抑制T细胞增殖,且呈剂量依赖性。因此,由抗体188定义的抗原被命名为豚鼠T淋巴细胞功能相关抗原-1,即gp TFA-1。抗体188的抑制程度在65%至85%之间,而其他三种针对豚鼠T细胞的抗体对T细胞增殖没有抑制作用。时间进程实验表明,gp TFA-1在T细胞活化的早期阶段起关键作用。只有在细胞培养开始时就存在抗体,才能实现最大程度的抑制;培养24小时后添加抗体不再有抑制作用。抗体188不会诱导T细胞有丝分裂。对从[35S]甲硫氨酸标记的T细胞母细胞的NP40裂解物中获得的免疫沉淀物进行二维分析(一维,IEF;二维,SDS-PAGE)表明,一种由两条非共价结合的多肽链组成的分子被特异性沉淀,其表观分子量分别为43,000和38,000。两个亚基均表现出广泛的电荷异质性,聚焦点的平均等电点分别为5.0和6.5。gp TFA-1分子在功能和结构特性上与最近描述的克隆型表达的T细胞糖蛋白具有惊人的相似性,这些糖蛋白在小鼠和人类系统中被证明参与T细胞的抗原识别。