Verastegui Manuela, Gilman Robert H, Arana Yanina, Barber Dylan, Velásquez Jeanette, Farfán Marilu, Chile Nancy, Kosek Jon C, Kosek Margaret, Garcia Hector H, Gonzalez Armando
Department of Microbiology, Universidad Peruana Cayetano Heredia, P.O. Box 5045, Lima, Peru.
Infect Immun. 2007 Nov;75(11):5158-66. doi: 10.1128/IAI.01175-06. Epub 2007 Aug 13.
The specific mechanisms underlying Taenia solium oncosphere adherence and penetration in the host have not been studied previously. We developed an in vitro adhesion model assay to evaluate the mechanisms of T. solium oncosphere adherence to the host cells. The following substrates were used: porcine intestinal mucosal scrapings (PIMS), porcine small intestinal mucosal explants (PSIME), Chinese hamster ovary cells (CHO cells), epithelial cells from ileocecal colorectal adenocarcinoma (HCT-8 cells), and epithelial cells from colorectal adenocarcinoma (Caco-2 cells). CHO cells were used to compare oncosphere adherence to fixed and viable cells, to determine the optimum time of oncosphere incubation, to determine the role of sera and monolayer cell maturation, and to determine the effect of temperature on oncosphere adherence. Light microscopy, scanning microscopy, and transmission microscopy were used to observe morphological characteristics of adhered oncospheres. This study showed in vitro adherence of activated T. solium oncospheres to PIMS, PSIME, monolayer CHO cells, Caco-2 cells, and HCT-8 cells. The reproducibility of T. solium oncosphere adherence was most easily measured with CHO cells. Adherence was enhanced by serum-binding medium with >5% fetal bovine serum, which resulted in a significantly greater number of oncospheres adhering than the number adhering when serum at a concentration less than 2.5% was used (P < 0.05). Oncosphere adherence decreased with incubation of cells at 4 degrees C compared with the adherence at 37 degrees C. Our studies also demonstrated that T. solium oncospheres attach to cells with elongated microvillus processes and that the oncospheres expel external secretory vesicles that have the same oncosphere processes.
猪带绦虫六钩蚴在宿主体内附着和侵入的具体机制此前尚未得到研究。我们开发了一种体外黏附模型试验,以评估猪带绦虫六钩蚴对宿主细胞的黏附机制。使用了以下底物:猪肠黏膜刮片(PIMS)、猪小肠黏膜外植体(PSIME)、中国仓鼠卵巢细胞(CHO细胞)、回盲部结直肠癌上皮细胞(HCT - 8细胞)和结直肠癌上皮细胞(Caco - 2细胞)。使用CHO细胞来比较六钩蚴对固定细胞和活细胞的黏附情况,确定六钩蚴孵育的最佳时间,确定血清和单层细胞成熟的作用,以及确定温度对六钩蚴黏附的影响。利用光学显微镜、扫描显微镜和透射显微镜观察黏附的六钩蚴的形态特征。本研究表明,活化的猪带绦虫六钩蚴在体外可黏附于PIMS、PSIME、单层CHO细胞、Caco - 2细胞和HCT - 8细胞。用CHO细胞最容易检测猪带绦虫六钩蚴黏附的可重复性。含>5%胎牛血清的血清结合培养基可增强黏附,与使用浓度小于2.5%血清时相比,黏附的六钩蚴数量显著更多(P < 0.05)。与37℃时的黏附情况相比,细胞在4℃孵育时六钩蚴的黏附减少。我们的研究还表明,猪带绦虫六钩蚴通过细长的微绒毛突起附着于细胞,并且六钩蚴会排出具有相同六钩蚴突起的外部分泌小泡。