Leishmania braziliensis Infection Enhances Toll-Like Receptors 2 and 4 Expression and Triggers TNF- α and IL-10 Production in Human Cutaneous Leishmaniasis

This study was carried out in accordance with the recommendations of Institutional Review Board of the Federal University of Bahia, Brazil, with written informed consent from all subjects. All subjects gave written informed consent in accordance with the Declaration of Helsinki. The protocol was approved by the Institutional Review Board of the Federal University of Bahia, Brazil (approval number 693.111). Human Blood Samples and Preparation of Peripheral Blood Cells Peripheral blood mononuclear cells (PBMC) were separated from heparinized venous blood by Ficoll-Hypaque gradient centrifugation. Cells were then washed in saline and resuspended in RPMI 1640 (supplemented with 5% of fetal calf serum, 100 U penicillin/mL, 100 ug streptomycin/ mL) (GIBCO BRL., Grand Island, NY, USA). Parasites An isolate of leishmania obtained from a skin lesion of a CL patient from Corte de Pedra (MHOM/BR/LTCP11245) was characterized as L. brazilensis using PCR and multicolus enzyme electrophoresis (Cupolillo et al., 1994). Parasites were initially grown in biphasic medium (NNN). After isolation, the parasite was cryopreserved in liquid nitrogen. The parasites selected for this study had not been previously passaged in liquid culture medium. After selection, the parasites were expanded in complete Schneider's medium (Aldrch Sigma, St. Louis, MO) supplemented with 10% fetal bovine serum (FBS) (Gilco BRL) and 2% sterile urine. For in vitro infection of PBMC, the promastigotes in the sta...
Source: Frontiers in cellular and infection microbiology - Category: Microbiology Source Type: research