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. 2021 Oct 28;218(12):e20192070. doi: 10.1084/jem.20192070

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© 2021 Isringhausen et al.

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Figure 8. — LCMV infection results in persistent functional impairment of the BM microenvironment to maintain HSC quiescence. (A) Experimental layout for CFSE label dilution analysis. (B) Representative dot plots for CFSE label dilution 21 d after transplantation of LSK cells into control (ctrl) uninfected or LCMV-cl13–infected mice, 56 dpi. CFSE dilution is shown for HSPC LSKCD48⁻ (left) and HSC LSKCD48⁻CD150⁺ (right) subsets. Numbers above dot plots indicate divisions demarcated by gates on the CFSE axis. (C) Quantification of the percentage of transplanted LSKCD48⁻CD150⁺ cells undergoing different numbers of divisions (div) based on dilution levels of CFSE (n = 7 recipient mice per time point from two independent experiments are shown in recipient control and LCMV-cl13–infected mice, 56 dpi). Statistics were analyzed by two-tailed Mann–Whitney U test with *, P < 0.05; **, P < 0.01; ***, P < 0.001; and ns, P > 0.05. (D) Experimental layout for long-term transplantation experiments. LSK cells from CD45.2 mice were transplanted into CD45.2/CD45.1 mice uninfected (CTRL) or infected with LCMV-cl13 (56 dpi) treated with PBS or a combination of a-IFNAR and anti-IFNγ. (E–I) Monthly values of CD45.2 chimerism in PB (E) myeloid Gr1⁺ (F), B220⁺ (G), CD4⁺ (H), and CD8⁺ (I) compartments. (J–L) CD45.2 chimerism in HSCs (LSKCD48⁻CD150⁺), multipotential progenitors (MPPs; LSKCD48⁺CD150⁻), common myeloid progenitors (CMPs; Lin⁻Sca-1⁻ckit⁺CD16/32⁻CD34⁺), granulocyte-monocyte progenitors (GMPs; Lin⁻Sca-1⁻ckit⁺CD16/32⁺CD34⁺), and common lymphoid progenitors (CLPs; Lin⁻Sca-1⁻ckit^+/loCD127⁺) in the BM at terminal analysis 4 mo after transplantation (n = 3 mice per group, one experiment). Statistical significance was analyzed by two-tailed ANOVA with Tukey’s multiple comparison test: *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, P > 0.05.