麻风杆菌特异性抗原酚糖脂-Ⅰ的单克隆抗独特型抗体的制备及初步鉴定

Production and Identification of Monoclonal Anti-idiotype Antibodies against Anti-Phenolic Glycolipid-I Antibody of Mycobacterium Leprae

作者将BALB/c鼠造成对人IgM分子的耐受状态后,再用从麻风病患者血清中分离的IgM型抗酚糖脂-Ⅰ(PGL-Ⅰ)抗体免疫,并经细胞融合,获得持续分泌单克隆抗独特型抗体(McAb_2)的细胞株4C_4。经鉴定,该McAb_2与兔抗PGL-Ⅰ抗体(Ab_1)的结合能被NT-O-BSA(PGL-Ⅰ的人工类似物)竞争抑制;McAb_2还能阻断纯化人Ab_1与NT-O-BSA的结合。以上均表明该McAb_2为具有PGL-Ⅰ抗原内影像作用的单克隆抗独特型抗体;McAb_2模拟抗原试验结果显示,该McAb_2具有代替PGL-Ⅰ用于麻风血清学诊断的可能性。

Hybridoma (4C_4) secretingmonoclone anti-idiotype antibody(McAb_2)against anti-phenolic glycolipid-Ⅰ (PGL-Ⅰ)antithey (Ab_1) was produced by fusionof SP2/0 myeloma cells and spleen cellsof syngeneic mice which had been previ-ously tolerant to human IgM 4C_4 mono-clone anti-idiotype antibody was identifi-ed with a series of experiments includingcompetitive and neutralizing inhibitionELISA. It was found that the binding ofMcAb_2 with rabbit anti-PGL-Ⅰ antibodycould be competitively inhibited by NT-O-BSA (synthetic analog of PGL-Ⅰ) andneutralized by polyclonal anti-PGL-Ⅰ anti-body derived from various origins (humanor rabbit); McAb_2 could block the bindingof purified human Ab_1 with NT-O-BSA.The assay of McAb_2 as mimic antigen de-monstrated that McAb_2 could substitutefor NT-O-BSA in leprosy serodiagnosis.These results show that anti-idiotype an-tibody produced by 4C_4 is a monocloneanti-idiotype antibody bearing internalimage of PGL-Ⅰ and possibly can be usedin lepyros serodaginosis.