Extended Data Fig. 1: Naive CD8+ T cells that recognize hepatocellular antigen are activated locally and expand, but do not develop effector function.
From: Dynamics and genomic landscape of CD8+ T cells undergoing hepatic priming
a, Schematic of the experimental setup. Five million Env28 TN cells were transferred into C57BL/6 × BALB/c F1 (WT) or HBV replication-competent transgenic (HBV Tg, C57BL/6 × BALB/c F1) recipients. Livers were collected and analysed 5 days after Env28 TN cell transfer and sera from the same mice were collected daily from day 0 to 5 after transfer. b, c, Absolute numbers of total (b) and IFNγ-producing (c) Env28 T cells in the livers of the indicated mice. d, ALT levels detected in the sera of the indicated mice at the indicated time points. n = 4. e, Schematic of the experimental setup. Five million Cor93 TN cells were transferred into C57BL/6 (WT) or MUP-core recipients. Mice were splenectomized and treated with anti-CD62L antibody 48 h or 4 h before cell transfer, respectively. Untreated wild-type mice that received 5 × 106 Cor93 TN cells were used as controls. Where indicated, mice were injected with 2.5 × 105 infectious units of non-replicating rLCMV-core 4 h before Cor93 TN cell transfer. Liver-draining lymph nodes68 (dLN) and non-draining inguinal lymph nodes (ndLN) were collected at 4 h and 1 day after transfer. f, Representative flow cytometry plots 4 h after Cor93 TN cells transfer. Numbers indicate the percentage of cells within the indicated gate. g, h, Quantification of the absolute numbers of cells recovered from the ndLN (g) and dLN (h) of the indicated mice 4 h and 1 day (d1) after Cor93 TN cell transfer. n = 3. i, Confocal immunofluorescence micrographs of liver sections from wild-type mice, wild-type mice transduced with rLCMV-core, MUP-core mice, and R26-ZsGreen mice injected with 2.5 × 105 infectious units of non-replicating rLCMV-cre. Scale bars, 100 μm. Note that, because HBV core protein did not accumulate at detectable levels in Kupffer cells and hepatic dendritic cells after rLCMV-core injection, we confirmed the tropism of this vector by injecting rLCMV-cre into R26-ZsGreen mice, which express the fluorescent protein ZsGreen after Cre-mediated recombination. j, MFI of CD69 expression on Cor93 T cells in the liver, blood, lung and bone marrow of the indicated mice 4 h after Cor93 TN cell transfer. n = 4. Data are mean ± s.e.m. and representative of at least three independent experiments. ***P < 0.001, two-tailed t-test (b, c) or one-way ANOVA with Bonferroni post-test (g, h, j). Mouse drawings were adapted from ref. 69.
