Figure 2.

Effects of L. infantum infection on BMDC CD40 and CD86 costimulatory molecule expression. A: BMDCs plated at 2 × 10⁶ cells/well were infected with L. infantum promastigotes in a 10:1 (parasites/cell) ratio for 4 hours. Cells were immediately recovered or left in culture for an additional 24 hours after the end of the infection. As a control, BDMCs were stimulated with 1 μg/ml LPS for 4 and 24 hours. The mRNA levels were assessed by qPCR for CD40 and CD86. Gene expression is indicated as mean log₂ values of fold changes relative to untreated cells. B: BMDCs were treated with 1 μg/ml LPS or pretreated for 1 hour with LPS and then infected with L. infantum promastigotes in a 10:1 (parasites/cell) ratio, (LPS → Infection). After 4 hours of culture, cells were harvested, and the CD40 and CD86 mRNA levels were assessed by qPCR. Gene expression is indicated as mean log₂ values of fold changes relative to LPS-treated cells. C: Cells were infected, treated with 1 μg/ml LPS or pretreated during 1 hour with LPS and then infected (LPS → Infection). After 24 hours, the surface expression of CD40 and CD86 was measured on gated CD11c⁺ cells by flow cytometry. As negative control, cells were incubated in RPMIc (DCs) for a similar time. Each value represents the mean ± SD from three independent experiments. **P < 0.01.