Kucknoor Ashwini S, Mundodi Vasanthakrishna, Alderete John F
Department of Microbiology and Immunology, University of Texas Health Science Center at San Antonio, TX, USA.
Cell Microbiol. 2007 Nov;9(11):2586-97. doi: 10.1111/j.1462-5822.2007.00979.x. Epub 2007 Jun 21.
We showed recently that contact of human vaginal epithelial cells (VECs) by Trichomonas vaginalis and incubation with trichomonad proteins in conditioned medium induced expression of VEC genes. We performed 2-D SDS-PAGE followed by MALDI-TOF to identify the major secreted proteins. Based on protein abundance and separation of spots in 2-D gels, 32 major secreted proteins were examined, which gave 19 proteins with accession numbers. These proteins included known secreted cysteine proteinases. In addition, other secreted proteins were enzymes of carbohydrate metabolism, adhesin protein AP65, heat shock proteins, thioredoxin reductase and coronins. We confirmed that the secreted trichomonad proteins induced expression of VEC genes, including interleukin 8 (IL-8), COX-2 and fibronectin. Purified AP65 added to VECs had a pronounced effect only on IL-8 gene expression, which was inhibited in the presence of 12G4 monoclonal antibody to AP65. Moreover, AP65 expressed episomally within epithelial cells was found to enhance the expression of IL-8 and COX-2. This may be the first report of analysis of the secreted proteins of T. vaginalis and of the host epithelial cell response to these proteins and to the prominent adhesin AP65.
我们最近发现,阴道毛滴虫与人类阴道上皮细胞(VECs)接触并在条件培养基中与滴虫蛋白一起孵育可诱导VEC基因表达。我们进行了二维SDS-PAGE,随后进行MALDI-TOF以鉴定主要的分泌蛋白。根据二维凝胶中蛋白质的丰度和斑点分离情况,检测了32种主要分泌蛋白,其中19种蛋白有登录号。这些蛋白包括已知的分泌型半胱氨酸蛋白酶。此外,其他分泌蛋白有碳水化合物代谢酶、粘附素蛋白AP65、热休克蛋白、硫氧还蛋白还原酶和冠蛋白。我们证实,分泌的滴虫蛋白可诱导VEC基因表达,包括白细胞介素8(IL-8)、COX-2和纤连蛋白。添加到VECs中的纯化AP65仅对IL-8基因表达有显著影响,在存在AP65的12G4单克隆抗体时其表达受到抑制。此外,发现在上皮细胞中游离表达的AP65可增强IL-8和COX-2的表达。这可能是关于阴道毛滴虫分泌蛋白以及宿主上皮细胞对这些蛋白和突出粘附素AP65反应分析的首篇报道。