Extended Data Fig. 8: vCD44BP–CD44 interactions in stromal cells but not hematopoietic cells affect the generation of antiviral CD8 T cell responses.
From: Fibroblastic reticular cells direct the initiation of T cell responses via CD44
(a) Representative flow cytometry plot showing proportions of donor (CD45.2) and recipient (CD45.1) derived haematopoietic cells in spleens of chimeric mice generated using the CRISPR-Cas9 based approach. (b) Proportions of donor (CD45.2) and recipient (CD45.1) derived DCs in spleens of chimeric mice. (c) Representative histograms showing CD44 expression on DCs isolated from spleens of chimeric mice by mechanical dissociation. (d) Representative flow cytometry plots of CD8 T cells and (e) M45+ virus specific CD8 T cells from chimeric mice infected with MCMV, ∆vCD44BP (day 7 PI) or uninfected controls. (f) Number of CD8 T cells in spleens of chimeric mice infected with MCMV, ∆vCD44BP (day 7 PI) or uninfected controls. Data in (a–f) combined from two independent experiments (gScr→WT: MCMV n = 8, ∆vCD44BP n = 7, naïve n = 4; gCd44 → WT: MCMV n = 8, ∆vCD44BP n = 8, naïve n = 4; gScr→Cd44−/−: MCMV n = 8, ∆vCD44BP n = 8, naïve n = 3) and representative flow plots concatenated from one experiment (naïve n = 2; MCMV and ∆vCD44BP n = 4). (g) Representative histograms showing CD44 expression on splenic stromal fibroblastic cells (FC), endothelial cells (EC) and dendritic cells (DC) isolated using a stromal isolation protocol; representative of three independent experiments (naïve n = 6; MCMV and ∆vCD44BP n = 11). (h) Representative flow cytometry plots showing expression of CD44 and mCherry by splenic EC from uninfected or MCMV:mCherry-infected BALB/c mice (24 h PI). All mice were infected with 5 × 103 PFU, except in (h) where 1 × 104 PFU was used. Mean is shown in flow plots; graphs show mean ± SEM; significance tested by two-sided Mann-Whitney.
