Analysis using canine peripheral blood for establishing in vitro conditions for monocyte differentiation into macrophages for Leishmania chagasi infection and T-cell subset purification.

• Autor(a) principal: Viana, Kelvinson Fernandes
• Data de Publicação: 2013
• Outros Autores: Soares, Rodrigo Dian de Oliveira Aguiar, Roatt, Bruno Mendes, Resende, Lucilene Aparecida, Lemos, Denise da Silveira, Oliveira, Rodrigo Corrêa de, Martins Filho, Olindo Assis, Moura, Sandra Aparecida Lima de, Zanini, Marcos Santos, Araújo, Márcio Sobreira Silva, Reis, Alexandre Barbosa, Giunchetti, Rodolfo Cordeiro
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Institucional da UFOP
• Texto Completo: http://www.repositorio.ufop.br/handle/123456789/4327; https://doi.org/10.1016/j.vetpar.2013.08.014
• topic: Leishmania chagasi; Leishmaniasis visceral canine; Mcrobicidal analysis; Macrophage

Canine visceral leishmaniasis (CVL) is a parasitic disease endemic in many countries, anddogs present as the major natural reservoir of the parasite, Leishmania chagasi (syn. L.infantum). Biomarkers in the canine immune system is an important technique in thecourse of developing vaccines and treatment strategies against CVL. New methodologiesfor studying the immune response of dogs during Leishmania infection and after receivingvaccines and treatments against CVL would be useful. In this context, we used peripheralblood mononuclear cells (PBMCs) from healthy dogs to evaluate procedures related to (i)establishment of in vitro conditions of monocytes differentiated into macrophages infectedwith L. chagasi and (ii) purification procedures of T-cell subsets (CD4+and CD8+) usingmicrobeads. Our data demonstrated that after 5 days of differentiation, macrophages wereable to induce significant phagocytic and microbicidal activity after L. chagasi infectionand also showed increased frequency of parasitism and a higher parasite load. Although N-acetyl- _-d-glucosaminidase (NAG) levels presented similar levels of macrophage cultureand L. chagasi infection, a progressive decrease in myeloperoxidase (MPO) levels was ahallmark over 5 days of culture. High purity levels (>90%) of CD4 and CD8 T cells wereobtained on a magnetic separation column. We concluded that monocytes differentiatedinto macrophages at 5 days and displayed an intermediate frequency of parasitism andparasite load 72 h after L. chagasi infection. Furthermore, the purification system usingcanine T-lymphocyte subsets obtained after 5 days of monocyte differentiation provedefficient for CD4 or CD8 T-cell purification (≥90%). The in vitro analysis using L. chagasi-infected macrophages and purified T cells presented a prospective methodology that couldbe incorporated in CVL vaccine and treatment studies that aim to analyze the microbicidalpotential induced by specific CD4+and/or CD8+T cells.