Miller K M, Lustigman S, Brotman B, Andrus L, Weiss N, Hotze C, Prince A M
Lindsley F. Kimball Research Institute of the New York Blood Center.
Trop Med Parasitol. 1991 Jun;42(2):85-90.
Five murine monoclonal antibodies (Mab's) have been produced which are reactive with the surface of the third-stage larvae (L3) of Onchocerca volvulus. These were produced from a fusion performed after intrasplenic injection of 10 live O. volvulus L3. Hybridomas were first screened by Elisa using a PBS extract of O. volvulus female adult worms. Elisa negative wells were screened by IFA on whole formalin-fixed L3. Five Mab's were isolated which were reactive with the surface of L3, all were found to be of the IgM isotype. All five Mab's were cross-reactive with the surface of O. lienalis L3, but not with the L3 of Brugia malayi, B. pahangi, Dirofilaria immitis, or Acanthocheilonema viteae. One of the Mab's, OV3-9, reacted with an antigen common to many species of helminths. The other four were genus specific reacting only with O. volvulus and O. lienalis. All 5 Mab's reacted with L2 of O. volvulus, but not with the L4 by IFA on whole fixed larvae. Only Mab's OV3-8 and OV3-9 reacted with cryosections of adult O. volvulus by IFA indicating that the other 3 Mab's are specific for the surface of L2/L3 of the genus Onchocerca. None of the Mab's reacted with detergent dissociated L3 proteins in Western Blot analysis. The ability of L3 to bind the Mab's was abolished by incubating larvae in either proteinase K or trypsin but was unaffected by incubation of larvae in detergents, or by treatment with periodate, indicating the proteinaceous nature of the Mab specific epitopes. Anti-phosphorylcholine Mab (Gib-13) did not react with the surface of L3.(ABSTRACT TRUNCATED AT 250 WORDS)
已制备出五种鼠单克隆抗体（Mab's），它们可与盘尾丝虫第三期幼虫（L3）表面发生反应。这些抗体是在脾脏内注射10条活的盘尾丝虫L3后进行融合产生的。杂交瘤首先通过酶联免疫吸附测定（ELISA）进行筛选，使用盘尾丝虫成年雌虫的磷酸盐缓冲盐水（PBS）提取物。ELISA阴性孔通过间接荧光抗体试验（IFA）在整个福尔马林固定的L3上进行筛选。分离出五种与L3表面发生反应的Mab's，发现它们均为IgM同种型。所有五种Mab's均与链尾丝虫L3表面交叉反应，但与马来布鲁线虫、彭亨布鲁线虫、犬恶丝虫或旋盘尾丝虫的L3不发生反应。其中一种Mab's，即OV3 - 9，与许多蠕虫物种共有的一种抗原发生反应。其他四种则具有属特异性，仅与盘尾丝虫和链尾丝虫发生反应。通过对整个固定幼虫进行IFA检测，所有5种Mab's均与盘尾丝虫的L2发生反应，但不与L4发生反应。只有Mab's OV3 - 8和OV3 - 9通过IFA与盘尾丝虫成虫的冰冻切片发生反应，表明其他3种Mab's对盘尾丝虫属的L2/L3表面具有特异性。在蛋白质印迹分析中，没有一种Mab's与经去污剂解离的L3蛋白发生反应。通过在蛋白酶K或胰蛋白酶中孵育幼虫，L3与Mab's结合的能力被消除，但在去污剂中孵育幼虫或用高碘酸盐处理对其没有影响，这表明Mab特异性表位具有蛋白质性质。抗磷酸胆碱Mab（Gib - 13）不与L3表面发生反应。（摘要截短于250字）
