Speer C A, Wong R B, Blixt J A, Schenkel R H
J Parasitol. 1985 Feb;71(1):33-42.
Sporozoites of Eimeria tenella were incubated for 10, 20, or 30 min with parasite-specific monoclonal IgG antibody 3D3II from mice and then rinsed in a Tris-buffered glucose saline solution (TBGS). Some sporozoites were then incubated for 10, 20, or 30 min with ferritin- or colloidal gold-conjugated goat anti-mouse IgG antibody and then fixed in 2.5% glutaraldehyde and prepared for transmission (TEM) or scanning (SEM) electron microscopy. Other sporozoites that had been previously exposed to monoclonal antibody were prefixed with 0.25% glutaraldehyde, incubated with ferritin- or colloidal gold-conjugated anti-mouse IgG antibody and then fixed and prepared for TEM or SEM. Control preparations consisted of sporozoites exposed only to TBGS, monoclonal antibody 3D3II or to ferritin- or colloidal gold-conjugated anti-mouse IgG antibody. Capping of immune complexes occurred only on the surface of those sporozoites exposed to monoclonal antibody 3D3II followed by ferritin- or gold-conjugated antibody. Immune complexes moved laterally and posteriorly on the outer surface of the parasite plasma membrane to form a cap at the posterior end of the sporozoite. Capping did not occur in TBGS controls nor in sporozoites treated with monoclonal antibody 3D3II and prefixed in 0.25% glutaraldehyde before exposure to ferritin- or gold-conjugated antibody. Thus, capping of surface antigens did not occur in the presence of monoclonal 3D3II antibody only, whereas specimens exposed to both monoclonal and ferritin- or colloidal gold-conjugated antibodies were able to cap immune complexes.
将柔嫩艾美耳球虫的子孢子与小鼠来源的寄生虫特异性单克隆IgG抗体3D3II孵育10、20或30分钟,然后在Tris缓冲葡萄糖盐溶液(TBGS)中冲洗。接着,一些子孢子与铁蛋白或胶体金偶联的山羊抗小鼠IgG抗体孵育10、20或30分钟,然后用2.5%戊二醛固定,制备用于透射电子显微镜(TEM)或扫描电子显微镜(SEM)观察。其他先前已接触单克隆抗体的子孢子先用0.25%戊二醛预固定,再与铁蛋白或胶体金偶联的抗小鼠IgG抗体孵育,然后固定并制备用于TEM或SEM观察。对照样本包括仅暴露于TBGS、单克隆抗体3D3II或铁蛋白或胶体金偶联的抗小鼠IgG抗体的子孢子。免疫复合物的帽化仅发生在那些先接触单克隆抗体3D3II,然后再接触铁蛋白或金偶联抗体的子孢子表面。免疫复合物在寄生虫质膜外表面横向和向后移动,在子孢子后端形成帽。在TBGS对照中以及在用单克隆抗体3D3II处理并在接触铁蛋白或金偶联抗体之前用0.25%戊二醛预固定的子孢子中均未发生帽化。因此,仅在存在单克隆3D3II抗体时不会发生表面抗原的帽化,而同时接触单克隆抗体和铁蛋白或胶体金偶联抗体的样本能够使免疫复合物帽化。