Cell therapy with IL-10-producing group 2 innate lymphoid cells suppresses Graft-versus-Host disease

Abstract

IL-10 producing group 2 innate lymphoid cells (ILC210) have immunoregulatory functions, and limit harmful immune responses across various tissues. Despite their crucial roles in maintaining immune homeostasis, the cell therapy potential of human ILC210has not been demonstrated, due to both limited numbers in human peripheral blood and lack of definitive markers for identification. Here, we isolate and expand circulating human ILC210, and assess their cell therapy potential in a humanized model of Graft-versus-Host Disease (GVHD). Cell therapy with human ILC210decreased GVHD severity and prolonged survival of NOD-scidIL2Rγnull(NSG) mice. Adoptive transfer of ILC210inhibited pathogenic T cell proliferation and intestinal infiltration, and suppressed CD4+Th1 and CD8+Tc1 cells in an IL-4 and IL-10 dependent manner. Critically, increased proportions of ILC2s did not correlate with higher rates of cancer relapse in HSCT recipients, and adoptive transfer of ILC210did not compromise graft-versus-leukemic (GVL) effects in a humanized model. Finally, we identify CD49d and CD86 as novel markers that discriminate ILC210from conventional ILC2s. Collectively, these findings demonstrate the potential of harnessing ILC210in cell therapies for GVHD and other immune-driven pathologies.

Efficient and stable CRISPR/Cas9-mediated genome-editing of human type 2 innate lymphoid cells

“…To confirm our protocol would be effective at targeting other genes in ILC2s, we performed a pilot experiment with sgRNAs targeting IL10 , which we and others have linked to ILC2s immunoregulatory effects in the context of allergy ( 11 ), cell therapy approaches for transplantation ( 28 ) and Graft-versus-Host Disease (GvHD) ( 31 ). We observed IL-10 expression was efficiently downregulated at day 3 post-transfection using our protocol ( Figure S7 ), indicating the approach is effective at knocking out genes in different genomic positions.…”

“…Similarly, ILC2s were cultured in X-VIVO15 (Lonza) supplemented with 5% human AB serum (Sigma), 100U/mL Penicillin-Streptomycin (Gibco) and 1X GlutaMAX (Gibco) with recombinant human IL-2, IL-7, and IL-33 as described in Reid et al. ( 31 ). Cytokine analysis was performed pre and post IL-4 knockout by flow cytometry and cytometric bead array ( Figure 1 ).…”