Fig 4.
Whole-lung leukocyte and lymphocyte recruitment following C. neoformans infection. Whole-lung leukocytes were isolated using CD45+ antibody-conjugated magnetic beads from lung cell suspensions of inbred 7-week-old female C3H/HeN and CBA/J mice at day 14 and day 21 after intratracheal infection with 104 CFU of C. neoformans ATCC 24067. The total leukocyte number was determined by an automated cell counter, and leukocyte subsets were analyzed by Diff-Quik staining of cell suspensions on cytospin slides. CD45+ leukocytes were also stained with fluorochrome-labeled antibodies specific for CD4+ T-lymphocyte, CD8+ T-lymphocyte, and CD19+ B-lymphocyte subsets and analyzed by flow cytometry. The absolute number of cells was determined by multiplying the percentage of each cell type by the total number of leukocytes (n = 4 to 6 mice/strain/time point; *, P < 0.05; **, P < 0.01; ***, P < 0.001).