Peroxisome Proliferator-Activated Receptor-γ-Mediated Polarization of Macrophages in Leishmania Infection
Kinetics of PPAR expression and parasitic infection. BALB/c mice were intravenously injected with 107 stationary phase promastigotes of L. donovani 1S. At two-week intervals, groups () were sacrificed, and their liver and spleen were harvested. PPAR expression was determined by real-time RT-PCR as described in legend of Figure 1. The amount of parasites in the organs was determined by limiting dilution analysis according to the procedure of Titus and colleagues . Cells were plated into wells of 96-well plates at a range of concentrations, according to the number of red blood cells in the liver and splenocytes in the spleen suspensions and then incubated at 27°C to allow the parasites to transform from intracellular amastigotes into promastigotes. After 2-3 weeks of proliferation, the number of wells that shows parasite growth was scored under a microscope, and the L-Calc software for limiting dilution analysis (provided by Stem Cell Technology, Vancouver, Canada) was used to determine the frequency of parasite. Normalization was based on the number of red blood cells in the liver cultures (a) and the number of splenocytes in the spleen cultures (b).
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