Leishmania major Infection in Humanized Mice Induces Systemic Infection and Provokes a Nonprotective Human Immune Response
Figure 4
Flow cytometric analyses of human immune cells in the spleen affected by L. major infection.
(A) Analyses of naïve (CD27+CD45RA+) and memory (CD27+CD45RA−) CD8+ (top) and CD4+ T cells in the spleen with (3×106 L. major; n = 9) or without infection (n = 4). (B) CFSE-labelled spleen cells isolated from L. major-infected humanized mice (3×106 L. major; n = 3) and control humanized mice (HM; without in vivo L. major infection; n = 2) re-stimulated with Leishmania major antigen (SLA), PMA or ConA for 72 hours. Cell samples were then analyzed for CD4+ T cell (light grey), CD8+ T cells (grey) and B cells proliferation (dark grey). Percentage of human T cells of three individual humanized mice samples were determined in C. (D) Human cytokine release from spleen cells (isolated from L. major-infected (3×106) humanized mice; n = 3) restimulated with soluble L. major antigen (SLA), PMA or without restimulation after 72 hours. pb = peripheral blood. Significances between groups were analyzed in one-way (C) or two-way-Anova (A) (A* = p<0,05; A** = p<0,01; A**** = p<0,0001). Additionally significances between groups are marked with * (p<0,05), ** (p<0,01), and *** (p<0,001) analyzed with Tukey's Multiple Comparison Test (1-way-Anova) and Bonferroni posttest (2-way-Anova).