Therapeutic polyclonal human CD8+ CD25+ Fox3+ TNFR2+ PD-L1+ regulatory cells induced ex-vivo

Abstract

We report that polyclonal CD8regs generated in one week ex-vivo with anti-CD3/28 beads and cytokines rapidly developed suppressive activity in vitro sustained by TGF-β. In immunodeficient mice, these CD8regs demonstrated a markedly protective, IL-10 dependent activity against a xeno-GVHD. They expressed IL-2Rα/β, Foxp3, TNFR2, and the negative co-stimulatory receptors CTLA-4, PD-1, PD-L1 and Tim-3. Suppressive activity in vitro correlated better with TNFR2 and PD-L1 than Foxp3. Blocking studies suggested that TNF enhanced PD-L1 expression and the suppressive activity of the CD8regs generated. Unlike other polyclonal CD4 and CD8 Tregs, these CD8regs preferentially targeted allogeneic T cells, but they lacked cytotoxic activity against them even after sensitization. Unlike CD4regs, these CD8regs could produce IL-2 and proliferate while inhibiting target cells. If these CD8regs can persist in foreign hosts without impairing immune surveillance, they could serve as a practical remission-inducing product for the treatment of autoimmune diseases, graft-versus-host disease, and allograft rejection.

Selective Activation of Tumor Necrosis Factor Receptor II Induces Antiinflammatory Responses and Alleviates Experimental Arthritis

“…To investigate the effect of selective TNFRII activation on both CD4+ and CD8+ Treg cells, we genetically engineered a novel mouse TNFRII agonist, EHD2‐sc‐mTNF R2 , and showed that this fusion protein is capable of expanding both mouse and human CD4+ and CD8+ Treg cells. As expected, the in vitro expansion of both CD4+ and CD8+ Treg cells was strongly increased in the presence of the T cell growth factor IL‐2, which was previously shown to be essential for the generation of clinically useful human CD4+ and CD8+ Treg cells. Our data show that TNFRII and IL‐2 signaling synergistically induce the expansion of Treg cells in vitro.…”

“…TNFRII signaling-induced expansion of CD4+ and CD8+ Treg cells. TNFRII is expressed on CD4+ and CD8+ Treg cells (20,23,24,36) and contributes to the expansion and stabilization of CD4+ Treg cells (19)(20)(21)(22). We therefore investigated the impact of EHD2-sc-mTNF R2 on CD4+ and CD8+ Treg cell expansion.…”

Suppression of Murine Lupus by CD4+ and CD8+ Treg Cells Induced by T Cell–Targeted Nanoparticles Loaded With Interleukin‐2 and Transforming Growth Factor β

“…Regarding the latter, the immunotherapeutic potential of CD8+ Treg cells in SLE has not been examined thoroughly, although it is known that improved function of CD8+ Treg cells in human SLE is associated with disease remission . We have previously shown that IL‐2 and TGFβ can induce CD8+ cells to become Treg cells , with a protective effect in humanized mice . Notably, when we used both CD4+ and CD8+ Treg cells induced ex vivo with IL‐2 and TGFβ to suppress lupus‐like disease in BDF1 mice, the therapeutic effects were much stronger than when the mice were treated with CD4+ Treg cells alone, suggesting an important role of CD8+ Treg cells in suppressing lupus autoimmunity .…”

The TNF Receptor Superfamily in Co-stimulating and Co-inhibitory Responses

“…Several studies suggest the immune modulatory role of TNFR2 is primarily in Treg cells due to high TNFR2 expression in Foxp3 + Treg cells compared to naïve CD4 + T cell. The activation of TNFR2 is important for proliferation, survival and lineage stability of Treg cells, and the development of thymic Treg cells from Treg precursor cells (Chen et al, 2013; Horwitz et al, 2013; Mahmud et al, 2014). TNFR2 also limits CD8 + T cell mediated viral clearance and anti-tumor immunity.…”