Cordero Esteban M, Gentil Luciana G, Crisante Gladys, Ramírez José Luis, Yoshida Nobuko, Añez Nestor, Franco da Silveira José
Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, UNIFESP, Rua Botucatu 862, CEP 04023-062, São Paulo, Brazil.
Acta Trop. 2008 Jan;105(1):87-91. doi: 10.1016/j.actatropica.2007.08.004. Epub 2007 Aug 21.
Trypanosoma cruzi, the parasite causing Chagas' disease, relies on triatomines for its transmission. T. cruzi metacyclic trypomastigotes express GP82 and GP90, which are developmentally regulated surface proteins that have been implicated in host cell invasion. We used quantitative RT-PCR to quantify GP90 and GP82 mRNA levels expressed by T. cruzi in the digestive tract of experimentally infected Rhodnius prolixus at different times post infection. Translation of these transcripts was assessed by immunofluorescence using specific monoclonal antibodies against GP90 and GP82. We found that although GP82 and GP90 proteins were not detected in epimastigote cells by immunofluorescence, transcripts were present at lower levels. Increased levels of GP90 and GP82 transcripts and the appearance of these proteins on the parasite surface were accompanied by morphological differentiation from epimastigotes into metacyclic forms. Our data suggest that during in vivo metacyclogenesis there is a coordinated mechanism that links stabilization of GP90 and GP82 mRNAs with their translation.
克氏锥虫是引发恰加斯病的寄生虫,其传播依赖于锥蝽。克氏锥虫循环后期锥鞭毛体表达GP82和GP90,这两种蛋白是受发育调控的表面蛋白,与宿主细胞入侵有关。我们运用定量逆转录聚合酶链反应(qRT-PCR)来量化克氏锥虫在实验感染的南美锥蝽消化道中于感染后不同时间表达的GP90和GP82信使核糖核酸(mRNA)水平。使用针对GP90和GP82的特异性单克隆抗体通过免疫荧光评估这些转录本的翻译情况。我们发现,尽管通过免疫荧光在无鞭毛体细胞中未检测到GP82和GP90蛋白,但转录本以较低水平存在。GP90和GP82转录本水平的增加以及这些蛋白在寄生虫表面的出现伴随着从无鞭毛体到循环后期形态的分化。我们的数据表明,在体内循环后期发育过程中,存在一种协调机制,将GP90和GP82 mRNA的稳定与其翻译联系起来。