Fig. 2: CD4+ Teff cells limit CD8+ T cell dysfunction induced by hepatocellular priming.
a, Design: Env126 Teff cells were transferred at a 1:1 ratio (106:106) with naive Env28 T cells in HBV Tg mice (Co-T). Single transfer of Env28 and untreated mice (phosphate-buffered saline (PBS)) were used as controls. b, sALT in indicated groups of mice and time points. c, IHL numbers at day 5 after cell transfer. d, Frequency and numbers of Env28 T cells. e,f, Frequency (e) and numbers (f) of IFNγ-, TNF- and Grzm-B-producing Env28 T cells. g, Representative flow cytometry plots showing the frequency of control and Env28 T cells expressing Ki-67 at medium (med) and high (high) levels. h, Frequency of Ki-67hi Env28 T cells. i, Radar plot showing normalized MFI values of indicated markers on Env28 T cells. Asterisks refer to the comparison between Env28 and Co-T conditions. j, Immunohistochemical representative micrographs of liver sections: hematoxylin and eosin (H&E), cytokeratin-7 (bile ducts) and Ki-67 stainings. Scale bars, 100 μm. Bottom, confocal immunofluorescence micrographs of liver sections showing: DsRed+/+ Env28 T cells (red), CD38+ sinusoids (white), F4/80+ KCs. Scale bars, 200 μm. k, HBV DNA quantification by Southern blot. Bands indicate integrated transgene (TG), relaxed circular (RC), double-stranded linear (DS) and single-stranded (SS) HBV DNA. l, Representative HBcAg immunohistochemistry (IHC) micrographs of liver sections. Scale bars, 100 μm. m, Serum HBV DNA quantification. n, Env126 Teff cell numbers. o, MFIs of CD44, CD62L, T-bet and Ki-67 on hepatic Env126 Teff cells. p, Representative flow cytometry plots showing the frequency of Env126 Teff cells producing IFNγ and TNF, expressing CXCR3 and T-bet. Control CD4+ T cells are shown in black. n = 3 (PBS), 4 (Env28) and 5 (Co-T). Data are representative of five independents and are expressed as the mean ± s.e.m. Data in b and m were analyzed using two-way ANOVA with Bonferroni’s post hoc test. Data in all other graphs were analyzed using a two-tailed t-test or one-way ANOVA with Bonferroni’s post hoc test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. dLNs, draining lymph nodes.
