Fig 4. Confocal microscopy of mouse spleen mononuclear cells from mice infected with L. infantum.

Mice were infected with promastigote forms of L. infantum and sacrificed 30 days (A) or 45 days (B) later. Spleen mononuclear cells were obtained and enriched for CD45- CD71+ cells. Cells were incubated in 8-well chamber slides to adhere followed by staining with DAPI, anti-CD271, anti-Sca1 and rabbit anti-L. infantum IgG. Note in Merge clear images that point to the presence of amastigote forms stained with anti-leishmania FITC antibody associated with CD271+, Sca1+ cells. Arrows in Zoom from merge shows one of several round images that clearly suggest amastigote forms of the parasite. No staining with anti-leishmania-FITC was seen in all preparations using cells from non-infected mice (not shown).