Becker I, Pérez-Montfort R, Pérez-Torres A, Rondán-Zárate A, Montfort I, Pérez-Tamayo R
Departamento de Medicina Experimental, Facultad de Medicina U.N.A.M., Mexico City D.F., Mexico.
Exp Parasitol. 1996 Mar;82(2):171-81. doi: 10.1006/expr.1996.0022.
We produced a monoclonal antibody against a major cysteine proteinase of 30kDa from trophozoites of Entamoeba histolytica strain HM1:IMSS. The specificity of the monoclonal antibody was confirmed by specific inhibition of azocasein digestion and by electrophoretic analysis, in the presence of sodium dodecyl sulfate or on a substrate gel, of the antigen precipitated by the antibody. Immunofluorescent staining of trophozoites with the monoclonal antibody revealed heterogeneity in the intensity of whole cell fluorescence and subcellular localization of the stain. The latter was also observed in trophozoites, which were stained by conventional immunohistochemical methods, from experimental liver abscesses in hamsters. Ultrastructural analysis showed antigen distributed mainly in clear amorphous zones in the cytoplasm, which were not limited by a visible membrane. Proteinases are translocated from these compartments to phagocytic vacuoles after trophozoites ingest erythrocytes, suggesting that these regions might be a lysosomal equivalent of this primitive eukaryotic cell.
我们制备了一种针对溶组织内阿米巴HM1:IMSS株滋养体中一种30kDa主要半胱氨酸蛋白酶的单克隆抗体。通过对偶氮酪蛋白消化的特异性抑制以及在十二烷基硫酸钠存在下或在底物凝胶上对抗体沉淀抗原进行电泳分析，证实了该单克隆抗体的特异性。用该单克隆抗体对滋养体进行免疫荧光染色，显示全细胞荧光强度和染色的亚细胞定位存在异质性。在通过传统免疫组织化学方法染色的来自仓鼠实验性肝脓肿的滋养体中也观察到了后者。超微结构分析表明，抗原主要分布在细胞质中清晰的无定形区域，这些区域没有可见膜的限制。滋养体摄取红细胞后，蛋白酶从这些区室转运到吞噬泡，这表明这些区域可能是这种原始真核细胞的溶酶体等效物。
