L. (L.) amazonensis and L. (L.) infantum promastigotes were cultured in 199 medium (Sigma Aldrich, Burlington, MA, USA) supplemented with 0.005% hemin, 40 mM HEPES pH 7.4, 100 µM adenine, 4 mM sodium bicarbonate, 20 μg/mL gentamicin, and 10% FBS. Promastigotes of LV79 (MPRO/BR/72/M1841, obtained from the rodent Proechimys sp. from Pará State, Brazil) and PH8 (IFLA/BR/67/PH8, isolated from the sand fly Lutzomyia flaviscutellata from Pará State, Brazil) strains were obtained through cultivation of amastigotes derived from BALB/c mice lesions in medium 199 at 24 °C. Parasites were subcultured weekly to an initial density of 2 × 106 parasites/mL until the eighth passage. For that, BALB/c mice infected with LV79 and with PH8 were kept in our animal facility. Amastigotes were collected from the footpad lesions when promastigotes’ cultures reached 6–7 passages.
All experiments employed parasites at fourth day of culture (early stationary phase); macrophage infections employed day 4 and day 6 cultures.
L. (L.) infantum Ba262 strain (MCAN/BR/1989/BA262) was used as control for LPG assays.
All experiments employed parasites at fourth day of culture (early stationary phase); macrophage infections employed day 4 and day 6 cultures.
L. (L.) infantum Ba262 strain (MCAN/BR/1989/BA262) was used as control for LPG assays.
Free full text: Click here
