title: Apoptosis-inducing factor participation in bovine macrophage Mycobacterium bovis-induced caspase-independent cell death.
abstract: Mycobacterium tuberculosis complex species survive and replicate in phagosomes of the host cell. Cell death (CD) has been highlighted as one of the probable outcomes in this host-pathogen interaction. Previously, our group demonstrated macrophage apoptosis as a consequence of Mycobacterium bovis infection. In this study, we aimed to identify the contribution of apoptotic effector elements in M. bovis-induced CD. Bovine macrophages were either infected with M. bovis (multiplicity of infection, 10:1) or treated with an M. bovis cell extract (CFE). Structural changes compatible with CD were evaluated. Chromatin condensation was increased three times by the CFE. On the other hand, a terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) assay demonstrated that levels of DNA fragmentation induced by M. bovis and CFE were 53.7% +/- 24% and 38.9% +/- 14%, respectively, whereas control cells had a basal proportion of 8.9% +/- 4.1%. Rates of DNA fragmentation were unaffected by the presence of the pan-caspase inhibitor N-benzyloxycarbonyl-Val-Ala-Asp (z-VAD). Cells treated with 100 mug of CFE for 12 h had a fivefold decrease in the level of mitochondrial outer membrane permeabilization compared to that of untreated cells. Neither M. bovis infection nor CFE treatment induced activation of caspase 3, 8, or 9. Translocation of apoptosis-inducing factor (AIF) to the nucleus was identified in 32% +/- 3.5% and 26.3% +/- 4.9% of M. bovis-infected and CFE-treated cells, respectively. Incubation of macrophages with z-VAD prior to infection did not alter the percentage of cells showing AIF translocation. Our data suggest that M. bovis-induced CD in bovine macrophages is caspase independent with AIF participation.
authors.affiliation: Departamento de Microbiología e Inmunología, Facultad de Medicina Veterinaria y Zootecnia, Universidad Nacional Autónoma de México, Circuito Exterior S/N, Ciudad Universitaria, Coyoacán 04510, Mexico City, Mexico.
authors.forename: X
authors.initials: X
authors.lastname: Vega-Manriquez
authors.forename: Y
authors.initials: Y
authors.lastname: López-Vidal
authors.forename: J
authors.initials: J
authors.lastname: Moran
authors.forename: L G
authors.initials: LG
authors.lastname: Adams
authors.forename: J A
authors.initials: JA
authors.lastname: Gutiérrez-Pabello
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000276:immunology*; D051033:Apoptosis Inducing Factor / Q000502:physiology*; D020169:Caspases / Q000502:physiology; D002417:Cattle; D016923:Cell Death / Q000276:immunology; D002457:Cell Extracts; D002478:Cells, Cultured; D053938:DNA Fragmentation; D008264:Macrophages / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology*; D009163:Mycobacterium bovis / Q000276:immunology; D021381:Protein Transport / Q000276:immunology
chemical_list: D051033:Apoptosis Inducing Factor; D002457:Cell Extracts; D020169:Caspases