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Acanthamoeba spp. monoclonal antibody against a CPA2 transporter: a promising molecular tool for acanthamoebiasis diagnosis and encystment study

Published online by Cambridge University Press:  21 September 2020

Michele Martha Weber-Lima
Affiliation:
Laboratório de Imunoquímica, Departamento de Patologia Básica, Universidade Federal do Paraná, Curitiba-PR, Brazil
Bianca Prado-Costa
Affiliation:
Laboratório de Imunoquímica, Departamento de Patologia Básica, Universidade Federal do Paraná, Curitiba-PR, Brazil
Alessandra Becker-Finco
Affiliation:
Laboratório de Imunoquímica, Departamento de Patologia Básica, Universidade Federal do Paraná, Curitiba-PR, Brazil
Adriana Oliveira Costa
Affiliation:
Departamento de Análises Clínicas e Toxicológicas, Faculdade de Farmácia, Universidade Federal de Minas Gerais, Belo Horizonte-MG, Brazil
Philippe Billilad
Affiliation:
IPSIT, School of Pharmacy, University Paris-Saclay, Châtenay-Malabry, France
Cinthia Furst
Affiliation:
Departamento de Patologia, Centro de Ciências da Saúde, Universidade Federal do Espírito Santo, Vitória, ES, Brazil
Juliana Ferreira de Moura
Affiliation:
Laboratório de Imunoquímica, Departamento de Patologia Básica, Universidade Federal do Paraná, Curitiba-PR, Brazil
Larissa Magalhães Alvarenga*
Affiliation:
Laboratório de Imunoquímica, Departamento de Patologia Básica, Universidade Federal do Paraná, Curitiba-PR, Brazil
*
Author for correspondence: Larissa Magalhães Alvarenga, E-mail: lmalvarenga@ufpr.br; lmalvarenga@gmail.com

Abstract

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.

Type
Research Article
Copyright
Copyright © The Author(s), 2020. Published by Cambridge University Press

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Footnotes

*

Those authors contributed equally for the manuscript

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