Rosfjord E C, Mikhail K S, Rowland E C, Powell M R
Department of Zoological and Biomedical Sciences, Ohio University, Athens 45701.
J Parasitol. 1990 Oct;76(5):698-702.
Susceptible C3H/He mice were immunized with the avirulent Corpus Christi strain of Trypanosoma cruzi and subsequently infected with virulent Brazil stain organisms. Seventy days after infection sera were isolated and analyzed on western blots of electrophoretically separated T. cruzi antigens prepared from culture-form parasites (primarily epimastigotes). More than 25 bands were identified. The antibodies were fractionated by elution from various regions of western blots corresponding to average molecular weights of approximately greater than 130, 77, 70, 60, 48, or 38 kDa. Each of these antibody preparations was then incubated with strips of nitrocellulose containing all of the electrophoretically separated T. cruzi, and cross-reactivity was determined. Antibodies isolated from the 130-, 77-, and 70-kDa regions all cross-reacted with each other. Antibodies eluted from the 60-kDa region bound antigens in the 60-, 70-, and the 77-kDa regions. More importantly, antibodies eluted from every region bound antigens in the 70-kDa region. Conversely, antibodies eluted from this region bound to antigens in all of the other regions. These data indicate the presence of (a) common antigenic epitope(s) in T. cruzi infections in these mice that is predominantly found in the 70-kDa antigen-antibody complex on western blots.
将易感的C3H/He小鼠用克氏锥虫无毒的科珀斯克里斯蒂株进行免疫，随后用有毒的巴西株病原体进行感染。感染70天后分离血清，并在由培养型寄生虫（主要是无鞭毛体）制备的克氏锥虫抗原的蛋白质印迹上进行分析。鉴定出超过25条带。通过从蛋白质印迹的各个区域洗脱来分离抗体，这些区域对应的平均分子量约大于130、77、70、60、48或38 kDa。然后将每种抗体制剂与含有所有经电泳分离的克氏锥虫的硝酸纤维素条带一起孵育，并测定交叉反应性。从130 kDa、77 kDa和70 kDa区域分离的抗体彼此之间都有交叉反应。从60 kDa区域洗脱的抗体与60 kDa、70 kDa和77 kDa区域的抗原结合。更重要的是，从每个区域洗脱的抗体都与70 kDa区域的抗原结合。相反，从该区域洗脱的抗体与所有其他区域的抗原结合。这些数据表明，在这些小鼠的克氏锥虫感染中存在一种共同的抗原表位，在蛋白质印迹上主要存在于70 kDa抗原-抗体复合物中。
