Differential influence of gp63-like molecules in three distinct Leptomonas species on the adhesion to insect cells
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Parasites belonging to the Leptomonas genus have been used as model organisms for studying biochemical, cellular, and genetic processes unique to members of the Trypanosomatidae family. In the present study, the cell-associated and extracellular peptidases of three Leptomonas species, Leptomonas collosoma, Leptomonas samueli, and Leptomonas wallacei, were assayed and characterized by gelatin–sodium dodecyl sulfate polyacrylamide gel electrophoresis. All parasites released metallopeptidases, whereas no cell-associated proteolytic activity could be detected in the cellular extracts from L. collosoma. Western blotting probed with a polyclonal antibody raised against gp63 from Leishmania amazonensis revealed two major reactive polypeptides of apparent molecular masses of 63 and 52 kDa, with different intensities in cellular extracts and released proteins from the studied trypanosomatids. Flow cytometry and fluorescence microscopy analyses showed that the gp63-like molecules have a surface location. This is the first report on the presence of gp63-like molecules in L. collosoma, L. samueli, and L. wallacei. The pretreatment of L. samueli and L. wallacei with anti-gp63 antibody significantly diminished their association index to Aedes albopictus cell line (C6/36), suggesting a potential involvement of the gp63-like molecules in the interaction process of these insect trypanosomatids with the vector.
KeywordsInsect Cell Visceral Leishmaniasis Spend Culture Medium Insect Cell Line Chikungunya Virus
This study was supported by the Brazilian agencies: MCT/CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico), CEPG/UFRJ (Conselho de Ensino para Graduados e Pesquisas), FAPERJ (Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro), FUJB (Fundação Universitária José Bonifácio), and CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior).
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