Cytotoxic activity and degradation patterns of structural proteins by corneal isolates of Acanthamoeba spp

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dc.contributor.authorSant'ana, Viviane Peracini [UNIFESP]
dc.contributor.authorCarrijo-Carvalho, Linda Christian [UNIFESP]
dc.contributor.authorForonda, Annette Silva [UNIFESP]
dc.contributor.authorChudzinski-Tavassi, Ana Marisa
dc.contributor.authorFreitas, Denise de [UNIFESP]
dc.contributor.authorSouza de Carvalho, Fabio Ramos [UNIFESP]
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.contributor.institutionButantan Inst
dc.date.accessioned2016-01-24T14:39:48Z
dc.date.available2016-01-24T14:39:48Z
dc.date.issued2015-01-01
dc.description.abstractProteolytic enzymes secreted by trophozoites (amoebic secretome) are suggested as the main virulence factor involved in the severity of Acanthamoeba keratitis. the degradation profile of the main glycoprotein components of anterior and posterior portions of the cornea and the cytopathic effect of secretomes on endothelial cells by contact-independent mechanism were evaluated.Trophozoites were isolated primarily from corneal tissue samples (n = 11) and extracellular proteins were collected from axenic cell culture supernatants. the molecular weights of proteolytic enzymes were estimated by zymography. Enzymatic cleavage of laminin and fibronectin substrates by amoebic secretome was investigated and cluster analysis was applied to the proteolysis profiles. Primary cultures of endothelial cells were used in both qualitative and quantitative assays of cytophatogenicity.Differential patterns of proteolysis were observed among the Acanthamoeba secretomes that were analysed. the uniformity of laminin degradation contrasted with the diversity of the proteolysis profiles observed in the fibronectin substrate. Acanthamoeba secretome extracted from four clinical isolates was shown to be toxic when in contact with the endothelial cell monolayer (p < 0.01). Induction of apoptosis and membrane permeability, at different percentual values, were suggested as the main mechanisms that could induce endothelial cell death when in contact with amoebic secretome.Our results provide evidence that virulence factors secreted by Acanthamoeba trophozoites can be related to an increased pathogenicity pattern by an independent contact-trophozoite mechanism, through induction of endothelial cell death by apoptosis at a higher percentage than providing the lack of cell viability by the membrane-associated pore-forming toxin activity.en
dc.description.affiliationUniversidade Federal de São Paulo, Paulista Sch Med, Dept Ophthalmol & Visual Sci, São Paulo, Brazil
dc.description.affiliationButantan Inst, Biochem & Biophys Lab, São Paulo, Brazil
dc.description.affiliationUnifespUniversidade Federal de São Paulo, Paulista Sch Med, Dept Ophthalmol & Visual Sci, São Paulo, Brazil
dc.description.sourceWeb of Science
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.description.sponsorshipIDFAPESP: 2011/51626-1
dc.description.sponsorshipIDCNPq: 311612/2012-1
dc.description.sponsorshipIDFAPESP: 2012/15603-0
dc.format.extent65-75
dc.identifierhttp://dx.doi.org/10.1007/s00417-014-2783-3
dc.identifier.citationGraefes Archive for Clinical and Experimental Ophthalmology. New York: Springer, v. 253, n. 1, p. 65-75, 2015.
dc.identifier.doi10.1007/s00417-014-2783-3
dc.identifier.issn0721-832X
dc.identifier.urihttp://repositorio.unifesp.br/handle/11600/38571
dc.identifier.wosWOS:000347553200008
dc.language.isoeng
dc.publisherSpringer
dc.relation.ispartofGraefes Archive for Clinical and Experimental Ophthalmology
dc.rightsinfo:eu-repo/semantics/restrictedAccess
dc.rights.licensehttp://www.springer.com/open+access/authors+rights?SGWID=0-176704-12-683201-0
dc.subjectAcanthamoebaen
dc.subjectCorneaen
dc.subjectProteolysisen
dc.subjectFibronectinen
dc.subjectLamininen
dc.subjectendothelial cellen
dc.titleCytotoxic activity and degradation patterns of structural proteins by corneal isolates of Acanthamoeba sppen
dc.typeinfo:eu-repo/semantics/article
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