In vitro-induced Foxp3+ CD8+ regulatory T cells suppress allergic IgE response in the gut

Abstract

Several subsets of CD8⁺ T cells are known to have a suppressive function in different tissues and diseases in mice and humans. Due to the lack of a consensus on the phenotype of regulatory CD8⁺ T cells and very low frequency in the body, its clinical use as adoptive cellular therapy has not advanced much. In the present work, using DNA methyltransferase inhibitor 5-Aza-2'-deoxycytidine (Aza), we efficiently and stably differentiated naïve CD8⁺ T cells (CD8⁺ CD25^- CD44^- cells) into the CD8⁺ Foxp3⁺ regulatory CD8⁺ T cells (CD8 Tregs). We also generated OVA peptide_257-264 -specific CD8⁺ Foxp3⁺ Tregs. Compared with activated CD8 T cells, Aza plus TGF-β-induced CD8⁺ Foxp3⁺ Tregs showed significantly increased surface expression of CD39, CD73, CD122, CD62L, and CD103, and secreted TGF-β and suppressed the proliferation of effector CD4⁺ T cells. Interestingly, CD8⁺ Foxp3⁺ Tregs exhibited low expression of perforin and granzyme required for cytotoxic function. Analysis of chemokine receptors showed that TGF-β + Aza induced CD8⁺ Foxp3⁺ Tregs expressed gut-tropic chemokine receptors CCR6 and CCR9, and chemokine receptors CCR7 and CXCR3 required for mobilization into the spleen, lymph nodes, and gut-associated lymphoid tissues. Adoptive transfer of induced CD8⁺ Foxp3⁺ Tregs restored cholera toxin-induced breakdown of oral tolerance to OVA by regulating OVA-specific IgE and IgG1. Altogether, we showed an efficient method to generate antigen-specific CD8⁺ Foxp3⁺ Tregs, and the adoptive transfer of these cells induces oral tolerance by suppressing allergic response and maintaining intestinal homeostasis.

Keywords: CD8 Tregs; OVA; cholera toxin; intestinal homeostasis; oral tolerance.