Tregs epigenetically reprogrammed from autoreactive effector T cells mitigate established autoimmunity
Tyler R. Colson, James J. Cameron, Hayley I. Muendlein, Mei-An Nolan, Jamie L. Leiriao, James H. Kim, Alexander N. Poltorak, Xudong Li
Tyler R. Colson, James J. Cameron, Hayley I. Muendlein, Mei-An Nolan, Jamie L. Leiriao, James H. Kim, Alexander N. Poltorak, Xudong Li
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Research Article Immunology Inflammation

Tregs epigenetically reprogrammed from autoreactive effector T cells mitigate established autoimmunity

Abstract

Reprogramming autoreactive CD4+ effector T (Teff) cells into immunosuppressive Tregs represents a promising strategy for treating established autoimmune diseases. However, the stability and function of such reprogrammed Tregs under inflammatory conditions remain unclear. Here, we show that demethylation of core Treg identity genes in Teff cells yields lineage-stable effector T cell reprogrammed Tregs (ER-Tregs). A single adoptive transfer of ER-Tregs not only prevents autoimmune neuroinflammation in mice when given before disease onset but also arrests its progression when administered after onset. Compared with Foxp3-overexpressing Teff cells, induced Tregs from naive precursors, and endogenous Tregs, ER-Tregs provide superior protection against autoimmune neuroinflammation. This enhanced efficacy stems from their inherited autoantigen specificity and selectively preserved effector cell transcriptional programs, which together bolster their fitness in inflammatory environments and enhance their suppressive capacity. Our results establish epigenetic reprogramming of autoreactive Teff cells as an effective approach to generate potent, stable Tregs for the treatment of refractory autoimmune conditions.

Authors

Tyler R. Colson, James J. Cameron, Hayley I. Muendlein, Mei-An Nolan, Jamie L. Leiriao, James H. Kim, Alexander N. Poltorak, Xudong Li

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Figure 3

Foxp3 expression is required but not sufficient for the suppressor function of ER-Tregs.

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Foxp3 expression is required but not sufficient for the suppressor funct...
(A) Flow cytometry of IFN-γ and GM-CSF expression in CTVloCD4+ Teff cells, cocultured for 3 days with T cell–depleted splenocytes serving as antigen presenting cells (APCs), in the presence of MOG and the presence or absence of Foxp3Thy1.1 R26Cas9 ER-Tregs transduced with the retroviral vector (RV) expressing sgRNA targeting Foxp3 (sgFoxp3) or the nontargeting sgRNA (sgNT). (B) Flow cytometry of IL-10 and IL-17A expression in ER-Tregs as in A. (C) Flow cytometry of IFN-γ and GM-CSF expression in CTVloCD4+ Teff cells, cocultured for 3 days with APCs, in the presence of MOG and the presence or absence of Foxp3Thy1.1 ER-Tregs transduced with MigR1 empty vector or CD4+ Teff cells forced to express Foxp3 via retroviral transduction with MigR1-Foxp3. (D) Flow cytometry of CD25 and CTLA-4 expression in MigR1-ER-Tregs and MigR1-Foxp3 CD4+ Teff cells as in C. (E–G) EAE was induced via MOG/CFA immunization in CD45.2+ mice with or without adoptive transfer of CD45.1+Foxp3Thy1.1 MOG/CFA-primed CD4+ Teff cells reprogrammed into ER-Tregs or forced to express Foxp3 via retroviral transduction (Foxp3-RV-Teff), administered 1 day prior to immunization. Flow cytometry analyses were conducted at 16 dpi. n = 6 per group. (E) EAE disease curve. (F) Flow cytometry analysis of the frequencies of spinal cord CD4+ T cells. (G) Flow cytometry of Foxp3 expression in adoptively transferred ER-Tregs and Foxp3-RV CD4+ Teff cells in the spinal cord. (H) Flow cytometry of the frequencies and numbers of ER-Tregs or FRV-Tregs in the spinal cord. Data are shown as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, 1-way ANOVA and Holm-Šídák test in (A, C, E, and F) and unpaired 2-tailed t test in (B, D, G, and H).