Figure 2.

A1^−/− cells in chimaeric mice exhibit normal T cell immune responses in an acute influenza infection model. (a) Schematic overview of the influenza infection workflow: C57BL/6-Ly5.1xLy5.2 F1 mice were lethally irradiated and reconstituted with 1:1 mixed bone-marrow from C57BL/6-Ly5.1 wild-type and C57BL/6-Ly5.2 A1^−/− mice. Reconstituted mice were infected intra-nasally (i.n.) with 3000 pfu HKx31 influenza virus and lungs and spleens were analysed 8 days post infection by flow cytometry. (b) Leukocyte infiltrates were isolated from lungs of chimaeric mice by gradient centrifugation. The recovered cells were analysed by flow cytometry for tetramer-positive antigen-specific (NP⁺) CD8⁺ T cells, KLRG1⁺ short-lived CD8⁺ effector T cells and (c) central (CD44⁺CD62L⁺) as well as effector (CD44⁺CD62L⁻) memory-like CD8⁺ T cells. (d) Spleen cells from infected bone-marrow chimaeric mice were isolated and analysed for CD8⁺ NP⁺ cells, or central as well as effector memory-like CD8⁺ T cells. (e) CD4⁺ splenic T cells were further analysed for CD4⁺FoxP3⁻ naive (CD62L⁺CD44-), central or effector memory-like subsets. (f) CD4⁺ splenic T cells were analysed for FoxP3⁺ Treg cells, FoxP3⁻CXCR3⁺ Th1 cells, and FoxP3⁻CXCR5⁺PD-1⁺ Tfh cells. (g) Spleen cells from infected mice were analysed for Syndecan1⁺ plasma cells (PC) and B220⁺FAS⁺GL7⁺ germinal centre B cells (GC). Bars represent means±S.E.M. (n=4), irradiation-resistant Ly5.1/5.2 DP cells were excluded by electronic gating