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Parasitology ; 147(14): 1678-1688, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-32951614

RESUMO

Free-living amoeba of the genus Acanthamoeba are ubiquitous protozoa involved in opportunistic and non-opportunistic infection in humans, such as granulomatous amoebic encephalitis and amoebic keratitis. Both infections have challenging characteristics such as the formation of the resistant cysts in infected tissues, hampering the treatment and most usual diagnosis depending on time-consuming and/or low sensitivity techniques. The use of monoclonal antibodies presents itself as an opportunity for the development of more effective alternative diagnostic methods, as well as an important and useful tool in the search for new therapeutic targets. This study investigated the possibility of using a previously produced monoclonal antibody (mAb3), as a diagnostic tool for the detection of Acanthamoeba trophozoites by direct and indirect flow cytometry and immunofluorescence. Immunoprecipitation assay and mass spectrometry allowed the isolation of the antibody's target and suggested it is a transporter part of the CPA (cation: proton antiporter) superfamily. In vitro tests indicate an important role of this target in Acanthamoeba's encystment physiology. Our results support the importance of studying the role of CPA2 transporters in the context of acanthamoebiasis, as this may be a way to identify new therapeutic candidates.


Assuntos
Acanthamoeba/imunologia , Amebíase/diagnóstico , Proteínas de Protozoários/genética , Trocadores de Sódio-Hidrogênio/genética , Acanthamoeba/genética , Amebíase/parasitologia , Sequência de Aminoácidos , Anticorpos Monoclonais , Anticorpos Antiprotozoários , Citometria de Fluxo , Imunofluorescência , Estrutura Secundária de Proteína , Proteínas de Protozoários/química , Proteínas de Protozoários/metabolismo , Alinhamento de Sequência , Trocadores de Sódio-Hidrogênio/química , Trocadores de Sódio-Hidrogênio/metabolismo , Trofozoítos/genética , Trofozoítos/imunologia
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