Palmer G H, McGuire T C
J Immunol. 1984 Aug;133(2):1010-5.
The first demonstration of immune serum-mediated neutralization of Anaplasma marginale and identification of the involved surface antigens is presented. Anaplasma marginale initial bodies were purified from parasitized erythrocytes by using ultrasonic disruption and differential centrifugation. The initial bodies were morphologically intact, as determined by electron microscopy, were not agglutinated by anti-bovine erythrocyte sera, and were infective. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated minimal erythrocytic contamination in the purified initial bodies. Rabbit antisera against initial bodies completely neutralized the infectivity of purified initial bodies for splenectomized calves, whereas normal rabbit serum did not alter infectivity. Nine initial body proteins were surface radioiodinated by using lactoperoxidase, and five of these were precipitated by the neutralizing antibody (105,000, 86,000, 61,000, 36,000, and 31,000 apparent m.w.). The results demonstrate that initial body determinants capable of inducing neutralizing antibody are present. The identification of surface radiolabeled proteins immunoprecipitated by the neutralizing antibody provides candidates to be tested as protective immunogens in cattle.
本文首次展示了免疫血清介导的对边缘无形体的中和作用,并鉴定了相关的表面抗原。通过超声破碎和差速离心从感染的红细胞中纯化边缘无形体的原体。经电子显微镜测定,原体形态完整,不被抗牛红细胞血清凝集,且具有感染性。十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)显示纯化的原体中红细胞污染极少。针对原体的兔抗血清完全中和了纯化原体对脾切除小牛的感染性,而正常兔血清则不改变感染性。使用乳过氧化物酶对九种原体蛋白进行表面放射性碘化,其中五种被中和抗体沉淀(表观分子量分别为105,000、86,000、61,000、36,000和31,000)。结果表明存在能够诱导中和抗体的原体决定簇。对被中和抗体免疫沉淀的表面放射性标记蛋白的鉴定为作为牛保护性免疫原进行测试提供了候选物。