Burns G F, Boyd A W, Beverley P C
J Immunol. 1982 Oct;129(4):1451-7.
Two independently produced monoclonal antibodies, UCHT1, an IgG1 antibody, and OKT3, an IgG2 antibody, which specifically identify all human peripheral T lymphocytes, were shown to possess similar functional properties. Both were highly mitogenic to peripheral blood T lymphocytes, and UCHT1 blocked specific T lymphocyte killing of autologous B lymphoblasts when present during either the induction or the cytotoxic phase of the reaction; this, too, is a property reported for OKT3 in an analogous situation. Both antibodies also inhibited the 6-day proliferative response to soluble antigens and mixed lymphocyte culture, although this may be attributable to their strong mitogenicity earlier in culture. These properties were not found with other monoclonal antibodies directed against human T lymphocytes, and suggested that UCHT1 and OKT3 recognize important functional structure(s) on the T cell surface. The identity of the antigen recognized by these two antibodies was demonstrated by 2-directional co-capping, by cold antibody inhibition of binding of radiolabeled antibody, and by immunoprecipitation and two-dimensional gel analysis of labeled T cell surface proteins. Both UCHT1 and OKT3 bound with similar affinities (Ka approximately 2 X 10(9) M-1) to the same number of binding sites (2.8 X 10(4) sites/activated T cell), and both precipitated the same glycoprotein complex with a m.w. of approximately 19,000 in the presence of reducing agent.
两种独立制备的单克隆抗体,即IgG1抗体UCHT1和IgG2抗体OKT3,它们能特异性识别所有人类外周血T淋巴细胞,已被证明具有相似的功能特性。二者对外周血T淋巴细胞都有很强的促有丝分裂作用,并且在反应的诱导期或细胞毒性期存在时,UCHT1会阻断自体B淋巴母细胞的特异性T淋巴细胞杀伤作用;在类似情况下,OKT3也有此特性报道。两种抗体还抑制了对可溶性抗原和混合淋巴细胞培养的6天增殖反应,不过这可能归因于它们在培养早期的强烈促有丝分裂作用。针对人类T淋巴细胞的其他单克隆抗体未发现这些特性,这表明UCHT1和OKT3识别T细胞表面重要的功能结构。通过双向共帽法、冷抗体对放射性标记抗体结合的抑制作用以及对标记的T细胞表面蛋白进行免疫沉淀和二维凝胶分析,证明了这两种抗体识别的抗原相同。UCHT1和OKT3以相似的亲和力(Ka约为2×10⁹ M⁻¹)结合到相同数量的结合位点(2.8×10⁴个位点/活化T细胞),并且在还原剂存在的情况下,二者都沉淀出分子量约为19,000的相同糖蛋白复合物。