Constitutive induction of intestinal Tc17 cells in the absence of hematopoietic cell-specific MHC class II expression
Version of Record online: 25 AUG 2013
© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
European Journal of Immunology
Volume 43, Issue 11, pages 2896–2906, November 2013
How to Cite
Rubino, S. J., Geddes, K., Magalhaes, J. G., Streutker, C., Philpott, D. J. and Girardin, S. E. (2013), Constitutive induction of intestinal Tc17 cells in the absence of hematopoietic cell-specific MHC class II expression. Eur. J. Immunol., 43: 2896–2906. doi: 10.1002/eji.201243028
- Issue online: 20 NOV 2013
- Version of Record online: 25 AUG 2013
- Accepted manuscript online: 23 JUL 2013 08:44AM EST
- Manuscript Accepted: 19 JUL 2013
- Manuscript Revised: 13 JUN 2013
- Manuscript Received: 27 SEP 2012
- Canadian Institute of Health Research (CIHR)
- Crohn's and Colitis Foundation of Canada
- Banting and Best Graduate Scholarship from CIHR
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Figure S1. Representative back-gating of CD8+ and CD4+ LP T cells. Lamina Propria Lymphocytes were gated by FSC-A and SSC-A, then the single cells were gated on by SSC-W. Next, dead cells were excluded by live-dead aqua staining and LPLs were gated on TCRb+ cells and then sub-gated into CD4+ and CD8+ populations.
Figure S2. CD4+, CD8+ and MHCII+ cell characterization in MHCII-/-gWT mice. A-B) Gated on live TCRb+ lymphocytes, dot plots depict the number of CD4+ and CD8+ T cells in the lamina propria of WT and MHCII-/- mice (A) or WTgWT and MHCII-/-gWT chimeric mice (B). C) Expression of MHC class II by flow cytometry on live lymphocytes in the spleen and lamina propria of WTgWT and MHCII-/-gWT chimeric mice. Dot plots represent one representative of 3 to 5 independent experiments.
Figure S3. Number of IL-17+ and IFNg+ CD4 T cells after infection with C. rodentium. The bar graphs depict the number of cells per cecum of IL-17A+ (A) and IFNg+ (B) CD4+ T cells in WTWT and MHCII-/-WT mice (average of 3 to 5 experiments). Statistical analysis was performed using a two-tailed student's t-test where: * = p < 0.05, ns = not significant.
Figure S4. Intracellular IL-22 expression in CD4+TCRb+ LPLs. A-B) Gated on live CD4+TCRb+ lymphocytes, dot plots depict the percentage of IL-17+ and IL-22+ CD4+T cells in the lamina propria of WT WT and MHCII-/-WT mice infected with C. rodentium for 4 days (A) or infected with Salmonella enterica serovar Typhimurium for 1 day (B). One representative of two independent experiments.
Figure S5. Characterization of CIITA-/-WT chimeric mice. A) Domain organization of the Nod-like receptor CIITA: CARD = Caspase recruitment domain, TA = transcriptional activator domain, NBD = nucleotide binding domain and LRR = leucine-rich domain. B) Expression of MHC class II by flow cytometry on live lymphocytes in the lamina propria of WTWT and CIITA-/-WT chimeric mice. C) Survival curve of WTWT and CIITA-/-WT mice infected by oral gavage with 1×109 CFU of C. rodentium (4 to 6 mice per group).
Figure S6. Intracellular IL-22 expression in CD8+TCRb+ LPLs. Gated on live CD8+TCRb+ lymphocytes, dot plots depict the percentage of IL-17+ and IL-22+ CD8+T cells in the lamina propria of WT WT and MHCII-/-WT mice. One representative of two independent experiments.
Figure S7. CD44 expression on IL-17+, FOXP3+ and IFN+ CD8+ T cells in the lamina propria of MHCII-/-gWT mice. Dot plots depict the expression CD44, IL-17 (left), FOXP3 (middle) and IFN(right)on gated CD8+TCR+ LPLs isolated from uninfected MHCII-/-gWT mice. One representative of 3 independent experiments.
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