Purcell D F, McKenzie I F, Lublin D M, Johnson P M, Atkinson J P, Oglesby T J, Deacon N J
Department of Pathology, University of Melbourne, Parkville, Australia.
Immunology. 1990 Jun;70(2):155-61.
The non-lineage restricted human CD46 antigen, with two glycoproteins of 56,000 molecular weight (MW) and 66,000 MW, was defined using a panel of monoclonal antibodies (mAb) that included the E4.3 mAb to the HuLy-m5 antigen. Here the E4.3 mAb is used to show that two other human cell-surface molecules, membrane co-factor protein (MCP) of the complement system and trophoblast leucocyte-common antigen (TLX), are the same as HuLy-m5; thus, these three independently identified molecules are equivalently CD46. A mouse mAb to TLX (H316) and a specific rabbit antiserum to purified MCP (RA-MCP) blocked the binding of FITC-labelled E4.3 to the surface of human peripheral blood leucocytes (PBL). In sequential immunoprecipitation studies, E4.3 cleared all molecules detected by H316 and the RA-MCP antiserum. Immunoprecipitation from Chinese hamster ovary cells expressing transfected MCP cDNA showed that E4.3 detects both the mature 66,000 higher MW form of MCP and its 48,000 MW pro-MCP precursor, which lacks O-linked carbohydrate and bears only simple high-mannose-type N-linked carbohydrate. The IgG fraction of a polyclonal antiserum to purified MCP blocked factor I-mediated cleavage of C3b, whereas the E4.3 mAb did not. These data establish that three independently identified antigen systems are indeed the same: HuLy-m5, which shares a cross-reactive epitope with some primate retroviral gp 70 molecules and can be physically associated with class I major histocompatibility complex (MHC) chains in the cell membrane; MCP, of interest as a member of the regulators of complement activation gene family thought to protect autologous cells from complement activation; and TLX, a polymorphic molecule of interest for its potential role at the foeto-maternal tissue interface during pregnancy. Thus, the human CD46 antigen amalgamates the HuLy-m5, MCP and TLX cell-membrane glycoproteins.
使用一组单克隆抗体(mAb)定义了非谱系限制性人类CD46抗原,该抗原由两种分子量分别为56,000(MW)和66,000 MW的糖蛋白组成,其中包括针对HuLy-m5抗原的E4.3 mAb。在此,E4.3 mAb用于表明另外两种人类细胞表面分子,即补体系统的膜辅助蛋白(MCP)和滋养层白细胞共同抗原(TLX),与HuLy-m5相同;因此,这三个独立鉴定的分子等同于CD46。针对TLX的小鼠mAb(H316)和针对纯化MCP的特异性兔抗血清(RA-MCP)可阻断FITC标记的E4.3与人外周血白细胞(PBL)表面的结合。在连续免疫沉淀研究中,E4.3清除了H316和RA-MCP抗血清检测到的所有分子。从表达转染MCP cDNA的中国仓鼠卵巢细胞中进行免疫沉淀显示,E4.3可检测到MCP成熟的66,000更高MW形式及其48,000 MW的前体MCP,后者缺乏O-连接碳水化合物,仅带有简单的高甘露糖型N-连接碳水化合物。针对纯化MCP的多克隆抗血清的IgG部分可阻断因子I介导的C3b裂解,而E4.3 mAb则不能。这些数据证实,三个独立鉴定的抗原系统确实相同:HuLy-m5,它与一些灵长类逆转录病毒gp 70分子共享交叉反应表位,并且可以在细胞膜中与I类主要组织相容性复合体(MHC)链物理结合;MCP,作为补体激活调节基因家族的成员,被认为可保护自体细胞免受补体激活,因此受到关注;以及TLX,一种多态性分子,因其在妊娠期间胎儿-母体组织界面的潜在作用而受到关注。因此,人类CD46抗原融合了HuLy-m5、MCP和TLX细胞膜糖蛋白。