Single‐cell profiling of human CD127+ innate lymphoid cells reveals diverse immune phenotypes in hepatocellular carcinoma

Abstract Background and Aims Innate lymphoid cells (ILCs) are tissue‐resident lymphocytes that play critical roles in cytokine‐mediated regulation of homeostasis and inflammation. However, relationships between their immune phenotypic characteristics and HCC remain largely unexplored. Approach and Results We performed single‐cell RNA sequencing analysis on sorted hepatic ILC cells from human patients with HCC and validated using flow cytometry, multiplex immunofluorescence staining, and functional experiments. Moreover, we applied selection strategies to enrich ILC populations in HCC samples to investigate the effects of B cells on the immune reaction of inducible T cell costimulator (ICOS) + ILC2 cells. Dysregulation of ILCs was manifested by the changes in cell numbers or subset proportions in HCC. Seven subsets of 3433 ILCs were identified with unique properties, of which ICOS + ILC2a were preferentially enriched in HCC and correlated with poor prognosis. Mechanistically, we report that B cells, particularly resting naïve B cells, have a previously unrecognized function that is involved in inflammatory differentiation of ILC2 cells. B cell–derived ICOSL signaling was responsible for exacerbating inflammation through the increased production of IL‐13 in ICOS + ILC2a cells. Heat shock protein 70 (HSP70) genes Heat Shock Protein Family A Member 1A ( HSPA1A ) and Heat Shock Protein Family A Member 1B ( HSPA1B ) were highly expressed in ILC2s in late‐stage HCC, and targeting to ICOS and its downstream effector HSP70 in ILC2s suppressed tumor growth and remodeled the immunosuppressive tumor microenvironment. Conclusions This in‐depth understanding sheds light on B cell‐driven innate type 2 inflammation and provides a potential strategy for HCC immunotherapy.