Pathogenic T cells persist after reversal of autoimmune disease by immunosuppression with regulatory T cells
Article first published online: 25 MAR 2013
© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
European Journal of Immunology
Volume 43, Issue 5, pages 1286–1296, May 2013
How to Cite
Tu, E., Bourges, D., Gleeson, P. A., Ang, D. K. Y. and van Driel, I. R. (2013), Pathogenic T cells persist after reversal of autoimmune disease by immunosuppression with regulatory T cells. Eur. J. Immunol., 43: 1286–1296. doi: 10.1002/eji.201242771
- Issue published online: 25 APR 2013
- Article first published online: 25 MAR 2013
- Accepted manuscript online: 19 FEB 2013 02:46AM EST
- Manuscript Accepted: 13 FEB 2013
- Manuscript Revised: 16 JAN 2013
- Manuscript Received: 26 JUN 2012
- National Health and Medical Research Council of Australia and the University of Melbourne
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Figure S1. Generation of H+/K+ ATPase-specific iTreg cells. Purified CD4+CD25- T cells from A23.Foxp3GFP mice were cultured with irradiated splenocytes in the presence of IL-2, H/ Kα630–641 peptide, TGFβ and retinoic acid for 7 days as described in “Methods” to produce A23 iTreg cells and analysed by flow cytometry for Foxp3GFP and CD4 expression (A). “Before culture”, prior to culture to induce iTreg cells; “After culture”, after culture to induce iTreg cells; “After purification”, after cells were purified by flow cytometric sorting. Lymph node cells from H/Kα-/- mice (5 × 107) were transferred into irradiated BALB/cCrSlc mice with or without A23 iTreg cells (2 × 106). Mice that received lymph node cells from wildtype mice were used as normal controls (normal). Mice were analysed 8 weeks after transfer by haematoxylin and eosin staining of sections of stomach tissue (B), and quantitation of gastric pathology (C). Data pooled from 5 independent experiments and each dot represents data from one mouse. Mann-Whitney U test was used; bars, median. ***, P < 0.001.
Figure S2. Experimental strategies Two overall experimental strategies were employed in this work. In the “Cotransfer” experiments (A) gastritogenic T cells from H/Kα-/- mice and A23 iTreg cells were transferred at the same time into sublethally-irradiated BALB/cCrSlc mice as described in “Methods”. Analysis was performed 8 weeks after transfer. In experiments to test the “Therapeutic” effect of iTreg cells (B), autoimmune gastritis was induced, as above, by the transfer of gastritogenic T cells from H/Kα-/- mice into sublethally-irradiated BALB/cCrSlc mice. After 8 weeks mice then received one dose either A23 iTreg cells or polyclonal wildtype iTreg cells. The severity of gastritis in the treated and untreated mice was assessed either 10 weeks or 6 months after the administration of iTreg cells. In some cases (C and D), the pathogenic potential of the donor cells was assayed by repurifying the T cells from the H/Kα-/- donor mice and transferring them to athymic mice. The athymic mice were analysed 8 weeks later.
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