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Infect. Immun. doi:10.1128/IAI.01175-06
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

T. solium oncosphere adhesion to intestinal epithelial and Chinese Hamster Ovary (CHO) cells in vitro

Department of Microbiology, Universidad Peruana Cayetano Heredia, PO Box 5045, Lima, Peru; Department of International Health, Johns Hopkins University, Bloomberg School of Hygiene and Public Health, Baltimore, 615 N. Wolfe Street Room W#5515, Baltimore, MD 21205, USA; Faculty of Veterinary Science, the University of Melbourne, 250 Princes Highway, Werribe, Vic. 3030, Australia; Department of Pathology, Stanford University School of Medicine, Stanford, California 94305, USA; Cysticercosis Unit, Instituto de Ciencias Neurologicas. Jr. Ancash 1271, Barrios Altos, Lima, Perú; Public Health Section, School of Veterinary Medicine, Universidad Nacional Mayor de San Marcos, Apartado 03-5113, Lima 03, Peru; 7AB PRISMA, Calle Carlos Gonzales 251, San Miguel, Lima 32 - Peru

^* To whom correspondence should be addressed. Email: rgilman{at}jhsph.edu.

	Abstract

The specific mechanisms underlying T. solium oncosphere adherence and penetration in the host had not been studied. We developed an in vitro adhesion model assay to evaluate the mechanisms of T. solium oncospheres 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), epithelial cells from ileocecal colorectal adenocarcinoma (HCT-8), and epithelial cells from colorectal adenocarcinoma (CaCo2). 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, scanning, and transmission microscopy were used to observe adhered oncosphere morphologic characteristics. This study showed in vitro adherence of activated T. solium oncosphere to PIMS, PSIME, monolayer CHO, CaCo2 and HCT 8 cells. Reproducibility of T. solium oncosphere adherence was most easily measured in CHO cells. Adherence was enhanced by serum-binding media with >5% fetal bovine serum and resulted in a significantly greater number of oncospheres adhering than in sera with a concentration less than 2.5% (p<0.05). Oncosphere adherence decreased with incubation of cells at 4°C versus 37°C. Our studies have also demonstrated that T. solium oncospheres attach to cells with elongated microvilli processes, and the oncospheres expel external secretory vesicles that have the same oncosphere processes.