Extended Data Fig. 4: IGRP-specific autoimmune CD8 T cell populations are phenotypically and transcriptionally distinct from IGRP-specific effector and memory CD8 T cells generated during acute infections.
From: An autoimmune stem-like CD8 T cell population drives type 1 diabetes
a, Experimental scheme showing generation of NRP-V7 tet+ effector and memory CD8 T cells during an acute Listeria infection in comparison to NRP-V7 tet+ autoimmune CD8 T cells in NOD mice. A Listeria monocytogenes strain was generated expressing the NRP-V7 mimotope (L.m. NRP-V7); host mice (Balb/c or NOD) were infected with L.m. NRP-V7 and effector and memory T cells from indicated tissues were analyzed at indicated time points using NRP-V7 tetramer. Effector and memory T cells were compared with NRP-V7+ autoimmune T cell populations from NOD mice and tissues as indicated. b, Top, identification of NRP-V7+ CD8 T cells in pancreas (effectors (Eff); black) at day 6-7 post infection of Balb/c mice or autoimmune NRP-V7+ CD8 T cells in pancreas of uninfected NOD mice (AI; red). Right, frequency of NRP-V7+ of CD8 T cells. Expression levels of KLRG1, CD127, TBET, and TCF1 in Eff and AI NRP-V7+ CD8 T cells from pancreas of Balb/c and NOD, respectively, are shown and quantified. Eff, n=4 (%NRP-V7+, %KLRG1+, %CD127+, TCF1 MFI) or n = 6 (TBET MFI); AI, n=3 (%NRP-V7+, %KLRG1+, %CD127+, TBET MFI) or n = 4 (TCF1 MFI). c, Expression of KLRG1 and CD127 of NRP-V7+ effectors from spleens of L.m.-infected Balb/c mice. b, c, Representative of 3 independent experiments. d, Frequency of KLRG1+ of NRP-V7+ CD8 T cells from pancreas (left) and spleen (right) of L.m.-infected Balb/c and NOD mice at day 6 post L.m. NRP-V7 infection compared to autoimmune (AI) NRP-V7+ CD8 T cells from NOD mice. Eff, n=3; NOD Eff, n = 4; NOD, n = 3 (pancreas), n=4 (spleen). Representative of 1 experiment. e, Top, identification of NRP-V7+ CD8 T cells in pancreatic lymph node (pLN) (memory (Mem); black) at day 21-28 post infection and autoimmune (AI) NRP-V7+ CD8 T cells in pLN of uninfected NOD mice (blue). Frequency of NRP-V7+ of CD8 T cells quantified on right. Expression profiles and frequencies of KLRG1, CD127, TCF1, CD62L, CD122, and CXCR3 of Mem and AI NRP-V7+ CD8 T cells from pLN of Balb/c and NOD, respectively. Mem, n = 7 (%NRP-V7+), n=6 (%TCF1+CD44+), n = 9 (%CD62L+), n = 3 (%CD127+, CD122 MFI, CXCR3 MFI); AI, n = 3 (NRP-V7+), n=7 (%TCF1+CD44+), n = 11 (%CD62L+), n=4 (%CD127+, CD122 MFI, CXCR3 MFI), n = 3 (%NRP-V7+). Representative of 3 independent experiments; %CD62L+ shows 3 combined experiments. Statistical testing conducted by Student’s two-tailed unpaired t-test. Data are mean ± SEM. f, Gene set enrichment analysis of RNA-seq data from IGRP-specific pLN TCF1hi CD8 T cells (see Fig. 2) compared to central memory CD8 T cells generated during acute LCMV infection; dataset from Mackay et al, 2016, GSE70813. NES, normalized enrichment score.
