Marot-Leblond A, Robert R, Aubry J, Ezcurra P, Senet J M
Laboratoire d'Immunologie-Parasitologie, UFR des Sciences Médicales et Pharmaceutiques, Angers, France.
FEMS Immunol Med Microbiol. 1993 Aug;7(2):175-86. doi: 10.1111/j.1574-695X.1993.tb00397.x.
Germ tube specific fractions of the dimorphic pathogenic fungus Candida albicans were fractionated according to their ability to link to fibrinogen. These fibrinogen binding factors were used as immunogens to prepare monoclonal antibodies (mAbs) with BALB/c mice. Among the resulting mAbs, one (mAb 3D9.3) was shown by indirect immunofluorescence to be specific to the surface of the mycelial phase of the C. albicans species. No labelling of the cell wall of any other Candida species was observed. This morphological shape specificity was confirmed by immunoblotting where a polydispersed high molecular mass component was identified. The molecular mass varied with the extraction procedure used; over 210 kDa with EDTA-2ME treatment, and ranging from 110 to 220 kDa after Zymolyase digestion. This phase-specific epitope was sensitive to proteolysis with pronase E, proteinase K and trypsin, but not to periodate treatment. Further purification of this material would allow further development of new serodiagnostic assays that might be more specific for invasive disease than currently available tests.
根据与纤维蛋白原结合的能力,对二态致病性真菌白色念珠菌的芽管特异性组分进行了分级分离。这些纤维蛋白原结合因子被用作免疫原,以制备BALB/c小鼠的单克隆抗体(mAb)。在所得的单克隆抗体中,其中一种(单克隆抗体3D9.3)通过间接免疫荧光显示对白色念珠菌菌丝体阶段的表面具有特异性。未观察到对任何其他念珠菌属细胞壁的标记。这种形态形状特异性通过免疫印迹得到证实,其中鉴定出一种多分散的高分子量成分。分子量随所用提取程序而变化;经EDTA-2ME处理后超过210 kDa,经溶菌酶消化后范围为110至220 kDa。这种阶段特异性表位对链霉蛋白酶E、蛋白酶K和胰蛋白酶的蛋白水解敏感,但对高碘酸盐处理不敏感。对该物质的进一步纯化将有助于进一步开发新的血清诊断检测方法,这些方法可能比目前可用的检测方法对侵袭性疾病更具特异性。