title: Induction of virulence factors, apoptosis, and cytokines in precision-cut hamster liver slices infected with Entamoeba histolytica.
abstract: Precision-cut liver slices (PCLS) are mainly used to evaluate hepatotoxicity and metabolism of chemicals, as well as to study mechanisms of liver damage and repair. However, recently they have been used as a system to study amoebic infections. The aim of this study was to validate this model as an alternative for experimental amoebic liver absess (ALA) in animals. To do this, the PCLS was analyzed for the expression of amoebapore and cysteine proteinases 1 and 5, three of the most studied virulence factors of Entamoeba histolytica, as well as the induction of apoptosis and cytokines production in response to the ex vivo infection. PCHLS were prepared with the Brendel-Vitron tissue slicer and then, infected with 200,000 trophozoites of E. histolytica. Samples were taken at 0, 6, 12, 18, and 24 h and compared to control non-infected slices. Morphological studies were performed in order to verify the infection; while apoptosis was studied by TUNEL and PAS techniques. The expression of cysteine proteinases (1 and 5), and amoebapore, was analyzed by real-time PCR. By using ELISA assays, the production of cytokines was also studied. PCHLS were found to be a reproducible infection system, and E. histolytica caused the expression of cysteine proteinases and amoebapore in infected slices. At the same time, trophozoites induce release of cytokines and apoptotic death of the hepatocytes close to them. PCHLS represent a new and suitable alternative model to study the pathogenesis of hepatic amoebiasis.
authors.affiliation: Centro de Investigación Biomédica del Noreste, Instituto Mexicano del Seguro Social, Administración de Correos No. 4, Apartado Postal 020, Colonia Independencia, Monterrey, NL, CP 64720, Mexico. pilarcarranza@cibinmty.net
authors.forename: Pilar
authors.initials: P
authors.lastname: Carranza-Rosales
authors.forename: María Guadalupe
authors.initials: MG
authors.lastname: Santiago-Mauricio
authors.forename: Nancy Elena
authors.initials: NE
authors.lastname: Guzmán-Delgado
authors.forename: Javier
authors.initials: J
authors.lastname: Vargas-Villarreal
authors.forename: Gerardo
authors.initials: G
authors.lastname: Lozano-Garza
authors.forename: Ezequiel
authors.initials: E
authors.lastname: Viveros-Valdez
authors.forename: Rocío
authors.initials: R
authors.lastname: Ortiz-López
authors.forename: Javier
authors.initials: J
authors.lastname: Morán-Martínez
authors.forename: A Jay
authors.initials: AJ
authors.lastname: Gandolfi
mesh_terms: D000826:Animal Testing Alternatives / Q000379:methods; D000818:Animals; D017209:Apoptosis; D006224:Cricetinae; D057056:Cysteine Proteases / Q000235:genetics / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism*; D004195:Disease Models, Animal; D004748:Entamoeba histolytica / Q000276:immunology / Q000472:pathogenicity*; D015971:Gene Expression Regulation, Enzymologic; D020287:In Situ Nick-End Labeling; D008099:Liver / Q000276:immunology / Q000469:parasitology* / Q000473:pathology; D008297:Male; D008647:Mesocricetus; D010503:Periodic Acid-Schiff Reaction; D060888:Real-Time Polymerase Chain Reaction; D037521:Virulence Factors / Q000032:analysis / Q000235:genetics / Q000378:metabolism*
chemical_list: D016207:Cytokines; D037521:Virulence Factors; D057056:Cysteine Proteases