Fig. 4: CD4+ T cells help CD8+ T cell function by licensing APCs in the liver.
a, Experimental design. b, sALT in indicated groups of mice and time points. c, IHL numbers at day 5 after cell transfer. d, Numbers of Cor93 T cells in the liver. e, Numbers of IFNγ+ and Grzm-B+ Cor93 T cells. f, Frequency of IFNγ+ Cor93 T cells. g,h, MFIs of indicated markers on Cor93 T cells. i, Liver immunohistochemical representative micrographs at day 5 after cell transfer: H&E, cytokeratin-7, Ki-67 stainings. Scale bars, 100 μm. j, Representative HBcAg IHC liver micrographs at day 5 after cell transfer. Scale bars, 100 μm. k,l, Numbers of hepatic Env126 and IFNγ+ Env126 CD4+ Teff cells. m, Frequency of IFNγ+ Env126 Teff cells. n, Heat map of normalized MFI (nMFI) of indicated markers on KCs and DCs at day 5 after cell transfer. MFI values were normalized separately (lowest value set to 0 and highest to 100). o, Experimental setup. Livers were analyzed 24 h after cell transfer. p, Rendered confocal immunofluorescence representative liver micrographs from co-transferred mice: DsRed+/+Env28 T cells (red), EGFP+/+Env126 Teff cells (green), CD38+ sinusoids (white), F4/80+ KCs (blue). Scale bars, 100 μm and 10 μm. q, Time-coded shapes of DsRed+/+Env28 T (red) and EGFP+/+Env126 Teff (green) cell tracks of Supplementary Video 1. r, Single-cell speeds and arrest coefficients of Env28 T cells non-interacting or engaged in three-cell clusters (interacting; cutoff, <2.5 µm min−1). s, Representative multiplex immunofluorescence liver image from a chimpanzee acutely infected with HBV at week 20 after infection: CD4+ T cells (green), CD8+ T cells (red), macrophages (KP1, white), nuclei (DAPI, blue). Scale bar, 10 µm. Individual channels are displayed in Supplementary Fig. 3. a–n, n = 3 (PBS), 4 (Cor93), 4 (Ctr Co-T), 4 (-SLOs Co-T). Data are representative of four independent experiments. o–r, n = 5. r, n = 82 tracks (not interacting), 28 tracks (interacting). Data are representative of at least two independent experiments. Data are expressed as the mean ± s.e.m. Data in b were analyzed using two-way ANOVA with Bonferroni’s post hoc test. Data in all other graphs were analyzed using two-tailed t-test or one-way ANOVA with Bonferroni’s post hoc test. *P < 0.05, **P < 0.01, ****P < 0.0001.
