Extended Data Fig. 5: CREM/ICER has no checkpoint role to limit effector function in CD8 T cells during persistent hepatotropic infection.
From: A liver immune rheostat regulates CD8 T cell immunity in chronic HBV infection
a, Strategy for generation of T cell-specific Icer deficient mice and targeting of the Icer locus among the Crem exons for integration of loxP sites, for details see material and method section. b, Expansion of CD8 T cells from Icerfl/fl and Cd4Cre x Icerfl/fl mice after 24 h stimulation in vitro (unpaired two-sided t-test p = 0.5814, n = 4). c,d, CD25 expression and INFγ production of CD8 T cells from the spleen of Icerfl/fl and Cd4Cre x Icerfl/fl mice after activation for 3 d in vitro followed by 4 h restimulation with PMA/Ionomycin (PI) or anti-CD3/CD28-coated beads (αCD3/28) or left in medium as control (ctrl) (two-way ANOVA with Sidak’s multiple comparison, n = 3). e,f, Monitoring of Ad-TTR-GOL-infected Icerfl/fl and Cd4Cre x Icerfl/fl mice via bioluminescence in vivo imaging and sALT measurements (two-way ANOVA with Šídák’s multiple comparison, n = 5). g, Liver and spleen CD8 T cells from Icerfl/fl and Cd4Cre x Icerfl/fl mice on d30 p.i. (Ad-TTR-GOL) (two-way ANOVA with Šídák’s multiple comparison, Icerfl/fl: n = 7, Cd4CrexIcerfl/fl: n = 9). h, Frequencies of liver antigen-specific multimer+ CD8 T cells in Icerfl/fl and Cd4Cre x Icerfl/fl mice on d30 p.i. (Ad-TTR-GOL) (unpaired two-sided t-test, Icerfl/fl: n = 7, Cd4CrexIcerfl/fl: n = 9). i, INFγ-expressing CXCR6+ CD8 T cells from Icerfl/fl and Cd4Cre x Icerfl/fl mice on d30 p.i. (Ad-TTR-GOL) after re-stimulation with OVA257-264 peptide (one-way ANOVA with Sidak’s multiple comparison, medium Padj=0.9467, OVA257-265 Padj=0.0084, Icerfl/fl: n = 7, Cd4CrexIcerfl/fl: n = 9). j, sALT levels after 108 IU Ad-HBV infection of ICERfl/fl or Cd4Cre x Icerfl/fl mice (two-way ANOVA with Šídák’s multiple comparison, n = 6). k, IFNγ production by liver HBcore-specific CD8 T cells from Cd4Cre x Icerfl/fl or littermate control mice after Cor93-100 peptide restimulation ex vivo (two-way ANOVA with Turkey’s multiple comparison, n = 6). n.a. = not analysed; b-f one out of ≥ two independent experiments shown; p≥0.05, * p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, not significant (n.s.). errors are shown as SD with mean.
