CD71+VISTA+ erythroid cells promote the development and function of regulatory T cells through TGF-β

Abstract

Cell-surface transferrin receptor (CD71⁺) erythroid cells are abundant in newborns with immunomodulatory properties. Here, we show that neonatal CD71⁺ erythroid cells express significant levels of V-domain Immunoglobulin (Ig) Suppressor of T Cell Activation (VISTA) and, via constitutive production of transforming growth factor (TGF)- β, play a pivotal role in promotion of naïve CD4⁺ T cells into regulatory T cells (Tregs). Interestingly, we discovered that CD71⁺VISTA⁺ erythroid cells produce significantly higher levels of TGF-β compared to CD71⁺VISTA⁻ erythroid cells and CD71⁺ erythroid cells from the VISTA knock-out (KO) mice. As a result, CD71⁺VISTA⁺ erythroid cells—compared to CD71⁺VISTA⁻ and CD71⁺ erythroid cells from the VISTA KO mice—significantly exceed promotion of naïve CD4⁺ T cells into induced Tregs (iTreg) via TGF-β in vitro. However, depletion of CD71⁺ erythroid cells had no significant effects on the frequency of Tregs in vivo. Surprisingly, we observed that the remaining and/or newly generated CD71⁺ erythroid cells following anti-CD71 antibody administration exhibit a different gene expression profile, evidenced by the up-regulation of VISTA, TGF-β1, TGF-β2, and program death ligand-1 (PDL-1), which may account as a compensatory mechanism for the maintenance of Treg population. We also observed that iTreg development by CD71⁺ erythroid cells is mediated through the inhibition of key signaling molecules phosphorylated protein kinase B (phospho-Akt) and phosphorylated mechanistic target of rapamycin (phospho-mTOR). Finally, we found that elimination of Tregs using forkhead box P3 (FOXP3)-diptheria toxin receptor (DTR) mice resulted in a significant expansion in the frequency of CD71⁺ erythroid cells in vivo. Collectively, these studies provide a novel, to our knowledge, insight into the cross-talk between CD71⁺ erythroid cells and Tregs in newborns. Our results highlight the biological role of CD71⁺ erythroid cells in the neonatal period and possibly beyond. The primary role of the red blood cells is to transport oxygen, but we know relatively little about the other functions they perform. Following maturation, red blood cells exit the bone marrow and enter blood circulation. Their immature counterparts are normally absent or in very low frequency in the blood of healthy adults. However, we showed previously that immature red blood cells are abundant in the spleens of neonatal mice and in human umbilical cord blood and that these cells possess immunological properties. In this report, we studied a subset of neonatal immature red blood cells that express a protein called V-domain Immunoglobulin (Ig) Suppressor of T Cell Activation (VISTA) on their surface. We found that the presence of VISTA enables the cells to repeatedly produce the regulatory cytokine TGF-β. TGF-β induces a subset of naïve lymphocytes—the CD4⁺ T cells—and converts them into regulatory T cells, also known as Tregs. Tregs modulate and suppress other immune cells. Our studies provide novel insights, to our knowledge, into the immunological role of immature red blood cells in newborns.