Chronic infection with LCMV-cl13 disrupts BM hematopoiesis and leads to long-term impairment of HSC function. (A and B) FC-based quantification of total BM cellularity (A) and Ter-119+ BM erythroid progenitors (B) after infections with 2 × 106 ffu LCMV-cl13. (C) Representative pictures of thick femoral slices of uninfected control (ctrl) or chronic LCMV-cl13 infection (7 dpi). (D–I) FC-based quantification of BM CD8 T cells (D), CD4 T cells (E), and B220+ B cells (F). (G) LK (Lin−c-kit+) progenitors. (H) LSK (Lin−Sca-1+c-kit+) progenitors. (I) HSCs (LSKCD150+CD48−) at different time points after infection. (J) Quantification of Ki67/DAPI cell cycle analysis of HSCs (n = ≥4 mice per group) during the course of infections with LCMV-cl13. Statistical significance was analyzed by two-tailed Mann–Whitney U test for A–J: *, P < 0.05; **, P < 0.01; ***, P < 0.001. (K) Focus-forming assay for LCMV-cl13 in the BM during the course of infections (n = 3–5 mice per time point). (L) Top: Schematic experimental layout for ELDA transplantation performed in at least four replicates per condition. Bottom: Linear regression analyses for the transplantation, with indicated numbers representing ELDA estimates for HSC functionality (n = 4–5 mice per group) and HSC dose. Numbers of transplanted mice were lower for 7 and 14 dpi, given the scarcity of HSCs (see Fig. S1).
