Medicine Faculty, National Autonomus University of Mexico, General Hospital of Mexico, Mexico City, Mexico.
Exp Parasitol. 2010 Jul;125(3):244-50. doi: 10.1016/j.exppara.2010.01.026. Epub 2010 Feb 6.
We have purified Gal/GalNAc lectin from Entamoeba histolytica by electroelution. The purified protein was used to immunize rabbits and obtain polyclonal IgG's anti-lectin. These antibodies were used as tools to analyze the expression and localization of the amoebic lectin in both virulent (vEh) and non-virulent (nvEh) variants of axenically cultured HM1:IMSS strain. vEh is able to induce liver abscesses in hamsters, whereas nvEh has lost this ability. In vitro, amoebic trophozoites from both variants equally express this protein as shown by densitometric analysis of the corresponding band in Western blots from lysates. In both types of trophozoites, the pattern of distribution of the lectin was mainly on the surface. We have also compared by immunohistochemistry the presence and distribution of lectin in the in vivo liver lesions produced in hamsters. In order to prolong the survival of nvEh to analyze both variants in an in vivo model, hamsters inoculated with nvEh were treated with methyl prednisolone. Our results suggest that the Gal/GalNAc lectin is equally expressed in both nvEh and vEh.
我们用电泳洗脱法从溶组织内阿米巴中纯化了 Gal/GalNAc 凝集素。纯化的蛋白质被用于免疫兔子，获得多克隆 IgG 抗凝集素。这些抗体被用作工具来分析在 axenically 培养的 HM1:IMSS 株的毒力（vEh）和非毒力（nvEh）变体中，阿米巴凝集素的表达和定位。vEh 能够在仓鼠中诱导肝脓肿，而 nvEh 已经失去了这种能力。在体外，两种变体的阿米巴滋养体同样表达这种蛋白，这一点可以通过 Western blot 分析裂解物中相应条带的密度来证明。在这两种滋养体中，凝集素的分布模式主要在表面。我们还通过免疫组织化学比较了在仓鼠体内产生的肝病变中凝集素的存在和分布。为了延长 nvEh 的存活时间，以便在体内模型中分析两种变体，用甲基强的松龙处理接种 nvEh 的仓鼠。我们的结果表明，Gal/GalNAc 凝集素在 nvEh 和 vEh 中表达水平相等。
