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Roles of the endosymbiont and leishmanolysin-like molecules expressed by Crithidia deanei in the interaction with mammalian fibroblasts
Authors:Filipe P Matteoli  Lívia O Santos  Carla Holandino  André LS Santos
Institution:a Laboratório de Estudos Integrados em Bioquímica Microbiana, Departamento de Microbiologia Geral, Instituto de Microbiologia Prof. Paulo de Góes (IMPPG), Bloco E-subsolo, Centro de Ciências da Saúde (CCS), Universidade Federal do Rio de Janeiro (UFRJ), Av. Carlos Chagas Filho, 373, Cidade Universitária, Rio de Janeiro, RJ 21941-902, Brazil
b Laboratório de Biologia Molecular e Doenças Endêmicas, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, RJ 21045-900, Brazil
c Departamento de Medicamentos-Faculdade de Farmácia, CCS, UFRJ, Brazil
d Laboratório de Bioquímica de Proteases, Departamento de Microbiologia Geral, IMPPG, CCS, UFRJ, Brazil
Abstract:Crithidia deanei is an insect trypanosomatid that harbors a bacterial endosymbiont in its cytoplasm. In this work, we have demonstrated the influence of the endosymbiont on the interaction of C. deanei with mammalian fibroblasts, also implicating the surface leishmanolysin-like molecules of C. deanei in this process. The wild strain of C. deanei expressed a higher amount (2-fold) of leishmanolysin-like molecules in the parasite surface than the aposymbiotic strain. The treatment of parasites with anti-leishmanolysin antibodies or the fibroblasts with purified leishmanolysin-like molecules from C. deanei significantly reduced the association index. The aposymbiotic strain of C. deanei presented interaction rates about 2- and 3-fold lower with fibroblasts than the endosymbiont-bearing counterpart after 1 and 2 h, respectively. However, the association indexes were similar after 3 and 4 h of interaction. Additionally, we observed a 2-fold increase in the association index after 24-96 h of parasite-fibroblast interaction when compared to the interaction process performed for 4 h, irrespective to the presence of the endosymbiont, suggesting that fibroblasts support multiplication and survival of C. deanei. Both parasite strains were able to induce fibroblast lysis. Interestingly, the wild strain led to a 2-fold increase in fibroblasts death in comparison to the aposymbiotic strain after 48-96 h. We also showed that both wild and aposymbiotic biotinylated live parasites recognized the same receptor in the fibroblast cells.
Keywords:Crithidia deanei  Monoxenous trypanosomatid  Endosymbiont  Fibroblast  Leishmanolysin  Cellular interaction
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