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Infection and Immunity, November 2007, p. 5158-5166, Vol. 75, No. 11
0019-9567/07/$08.00+0 doi:10.1128/IAI.01175-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.
Taenia solium Oncosphere Adhesion to Intestinal Epithelial and Chinese Hamster Ovary Cells In Vitro
Manuela Verastegui,1
Robert H. Gilman,1,2,7*
Yanina Arana,1
Dylan Barber,3
Jeanette Velásquez,1
Marilu Farfán,1
Nancy Chile,1
Jon C. Kosek,4
Margaret Kosek,2
Hector H. Garcia,1,5
Armando Gonzalez,6 and the Cysticercosis Working Group in Peru
Department of Microbiology, Universidad Peruana Cayetano Heredia, P.O. Box 5045, Lima, Peru,1 Department of International Health, Johns Hopkins University, Bloomberg School of Hygiene and Public Health, 615 N. Wolfe Street, Room W#5515, Baltimore, Maryland 21205,2 Faculty of Veterinary Science, University of Melbourne, 250 Princes Highway, Werribe, Victoria 3030, Australia,3 Department of Pathology, Stanford University School of Medicine, Stanford, California 94305,4 Cysticercosis Unit, Instituto de Ciencias Neurologicas, Jr. Ancash 1271, Barrios Altos, Lima, Peru,5 Public Health Section, School of Veterinary Medicine, Universidad Nacional Mayor de San Marcos, Apartado 03-5113, Lima 03, Peru,6 AB PRISMA, Calle Carlos Gonzales 251, San Miguel, Lima 32, Peru7
Received 26 July 2006/ Returned for modification 14 September 2006/ Accepted 20 June 2007
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°C compared with the adherence at 37°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.
* Corresponding author. Mailing address: Department of International Health, Johns Hopkins University, Bloomberg School of Hygiene and Public Health, 615 N. Wolfe Street, Room W#5515, Baltimore, MD 21205. Phone: (410) 614-3959. Fax: (410) 614-6060. E-mail: rgilman{at}jhsph.edu
Published ahead of print on 13 August 2007.
Infection and Immunity, November 2007, p. 5158-5166, Vol. 75, No. 11
0019-9567/07/$08.00+0 doi:10.1128/IAI.01175-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.
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