FIG. 2.

The glycoconjugate molecules of Leishmania major do not modulate DC activation in vivo. BALB/c mice infected in the footpad with CFSE-labeled WT, lpg2 AB, or lpg2⁻ (lpg2 knockout) L. major parasites were sacrificed at 3 days, the dLNs were collected and then digested with collagenase, and the total number of cells was enumerated by direct cell count using a hemocytometer (A). DCs were isolated from pooled dLN cells by positive selection using autoMACS column, and stained routinely with fluorochrome-labeled anti-CD11c MAb and other various cell surface markers (CD40, CD80, CD86, and MHC II). The stained cells were analyzed by flow cytometry to enumerate the total number of DCs (i.e., CD11c⁺ cells in panel B) or percentage of infected cells (i.e., CFSE^high CD11c⁺ cells in panel C). The expression of MHC class II, CD40, CD80, and CD86 molecules (D) and CD8α molecules (E) on both infected (CFSE^high) and total DCs were analyzed after gating on CD11c⁺ cells. Data presented are representative of three (A to C) and two (D) independent experiments with similar results. ND, not done. *, P < 0.05; **, P < 0.01.