抗恶性疟原虫有性期单克隆抗体的制备和鉴定  

PREPARATION AND IDENTIFICATION OF MONOCLONAL ANTIBODY AGAINST SEXUAL STAGE OF PLASMODIUM FALCIPARUM

作　　者：许学年[1] 冯正[1] 杨柏林[1] 瞿靖琦[1] 包意芳[1] 许永湘[1] 王聚君[1] 胡薇[1] 

机构地区：[1]中国预防医学科学院寄生虫病研究所

出　　处：《实用寄生虫病杂志》1998年第4期145-147,共3页Journal of Practical Parasitic Diseases

基　　金：国家自然科学基金;国家"九五"科技攻关项目

摘　　要：以培养的恶性疟原虫ＮＦ５４（３Ｄ７）株配子体蛋白抽提液及我国云南现场采集的恶性疟原虫细胞骨架分别免疫ＢＡＬＢ／ｃ小鼠。取免疫小鼠脾细胞与ＳＰ２／０骨髓瘤细胞融合，以ＩＦＡ法筛选出８株抗恶性疟原虫有性期ＭｃＡｂ杂交瘤细胞株。经免疫球蛋白类别鉴定，６株为ＩｇＧ１（Ｍ２Ａ１０Ｃ９、Ｍ２Ｃ１Ｂ８、Ｍ４Ｃ７Ｂ１０、Ｍ４Ｇ１２Ｃ１、Ｍ５Ｂ７Ｅ６和Ｍ６Ｅ１Ｇ１１），２株为ＩｇＭ（Ｍ４Ｄ７Ｆ７和Ｍ６Ｆ４Ｄ６）。其中３株ＭｃＡｂｓ（Ｍ４Ｃ７Ｂ１０、Ｍ４Ｄ７Ｆ７和Ｍ６Ｅ１Ｇ１１）的靶抗原定位于配子体以及大滋养体和裂殖体期无性体原虫；其余５株仅定位于配子体。经Ｗｅｓｔｅｒｎ印迹试验，ＭｃＡｂ所识别的蛋白区带各异（１６－１２０ｋＤ），与已发现的有性期特异性抗原相比较，３２ｋＤ抗原国内外尚未报道。各株ＭｃＡｂ与猴疟（Ｐ．ｃｙｎｏｍｏｌｇｉ）红内期、鸡疟（Ｐ．ｇａｌｉｎａｃｅｕｍ）子孢子和杜氏利什曼原虫前鞭毛体均无交叉反应。Hybridomas secreting monoclonal antibodies against sexual stage of Plasmodium falciparum were produced by fusing SP2/0 myeloma cells with spleen cells of BALB/c mice immunized with gametocyte stage protein extracts and insoluble parasitic cytoskeleton pellet, respectively Of eight McAbs, six were classified as IgG1 (M 2A 10 C 9, M 2C 1B 8, M 4C 7B 10 , M 4G 12 C 1, M 5B 7E 6, M 6E 1G 11 ), and the others were IgM (M 4D 7F 7, M 6F 4D 6) Three McAbs (M 4C 7B 10 , M 4D 7F 7, M 6E 1G 11 ) clearly reacted with sexual and asexual parasites based on IFA test Western blot analysis showed that the molecualr weights of antigens recognized by the eight McAbs varied from 16 kD to 120 kD The 32 kD antigen might be a new one compared with the sexual stage specific antigens being reported in the literature Neither of these McAbs showed to have cross reaction with erythrocytic stage of Plasmodium cynomolgi, sporozites of Plasmodium gallinaceum and promastigote of Leishmania donovani

关 键 词：恶性疟原虫 有性期 传播阻断抗原 单克隆抗体 

分 类 号：R978.6[医药卫生—药品] R392－33[医药卫生—药学]