Gaafar T, Ismail S, Helmy M, Afifi A, Guirguis N, el Ridi R
Department of Clinical Pathology, Faculty of Medicine, Cairo University, Egypt.
Parasitol Res. 1993;79(2):103-8. doi: 10.1007/BF00932254.
Schistosoma haematobium soluble egg antigens (SH SEAs) induce intense granulomas in human hosts that often culminate in severe disease. In an attempt to identify the SH SEA fractions that are responsible for pathology, we combined T-cell Western blotting and an in vitro model of granuloma formation. Whole SH SEAs were dotted onto nitrocellulose pieces or were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electrotransferred onto nitrocellulose paper. Horizontal strips bearing the separated antigens were solubilized in dimethylsulfoxide and precipitated in carbonate/bicarbonate buffer. Antigen-free and antigen-bearing particles were used to stimulate peripheral blood mononuclear cells (PBMCs) obtained from S. haematobium-infected patients and sex- and age-matched healthy controls to form granulomas in vitro. Whole SH SEA-bearing nitrocellulose particles elicited in vitro formation of granulomas by PBMCs from infected donors. The response was similar in sensitivity, specificity, and reproducibility to that evoked by SH SEA-bound polyacrylamide beads. The results obtained in samples form 30 patients and 10 controls tested with SH SEA-separated fractions revealed that SEA bands of 84,000, 63,000, 57,000, 55,000, 40,000, 30,000, and 28,000 Da elicited in vitro granuloma reactions by PBMCs of almost all infected patients. Conversely, separated soluble adult-worm antigens failed to stimulate PBMCs of infected patients to form granulomas. This study is the first to identify the SH SEA fractions that evoke in vitro granuloma formation and represents an initial step toward the development of an anti-urinary schistosomiasis pathology vaccine.
埃及血吸虫可溶性虫卵抗原（SH SEAs）可在人体宿主中引发强烈的肉芽肿反应，常导致严重疾病。为了确定导致病变的SH SEA组分，我们将T细胞蛋白质免疫印迹法与肉芽肿形成的体外模型相结合。将完整的SH SEAs点样到硝酸纤维素片上，或通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳进行分离，然后电转移到硝酸纤维素纸上。带有分离抗原的水平条带在二甲基亚砜中溶解，并在碳酸盐/碳酸氢盐缓冲液中沉淀。使用不含抗原和带有抗原的颗粒刺激从感染埃及血吸虫的患者以及性别和年龄匹配的健康对照者中获取的外周血单核细胞（PBMCs），以在体外形成肉芽肿。带有完整SH SEA的硝酸纤维素颗粒可诱导感染供体的PBMCs在体外形成肉芽肿。该反应在敏感性、特异性和可重复性方面与SH SEA结合的聚丙烯酰胺珠引发的反应相似。对30例患者和10例对照的样本用SH SEA分离组分进行检测的结果显示，分子量为84,000、63,000、57,000、55,000、40,000、30,000和28,000道尔顿的SEA条带可诱导几乎所有感染患者的PBMCs产生体外肉芽肿反应。相反，分离的可溶性成虫抗原未能刺激感染患者的PBMCs形成肉芽肿。本研究首次确定了可引发体外肉芽肿形成的SH SEA组分，是朝着开发抗泌尿血吸虫病病理疫苗迈出的第一步。
