Tumor infiltrating and peripheral CD4+ILT2+ T cells are a cytotoxic subset selectively inhibited by HLA-G in clear cell renal cell carcinoma patients

Abstract

ILT2 has recently been positioned as a major immune checkpoint in urologic cancers. In clear cell renal cell carcinoma (ccRCC), tumor-infiltrating CD8⁺ T cells expressing ILT2 are a highly cytotoxic cell population, distinct from PD1⁺ T cells, and whose function is inhibited by HLA-G⁺ targets. Here we report that ILT2 receptor can also be expressed by CD4⁺ T cells in urologic cancer patients. In the course of deciphering the role of these ILT2⁺CD4⁺ T cells, we found a statistical association between the tumor context and these T cells, and a positive correlation between the levels of peripheral and intra-tumoral CD4⁺ILT2⁺ T cells. Phenotypic analyses revealed that CD4⁺ILT2⁺ T cells express memory T cell (CD27^-CD28^-CD57⁺) and cytotoxicity (Tbet⁺Perforin⁺KLRG1⁺NKp80⁺GPR56⁺) markers, consistent with a CD4⁺CTL phenotype. Functional assays showed that ccRCC-infiltrating CD4⁺ILT2⁺ T cells indeed have high cytolytic properties and therefore function as proper CD4⁺CTLs, but are selectively inhibited by HLA-G⁺ targets. Clinical relevance was provided by immunohistochemical analyses on ccRCC tumor lesions with HLA-G⁺ HLA class II⁺ tumor cells next to CD4⁺ T cell infiltrates. Our findings provide evidence supporting that ILT2⁺ T cells constitute a reservoir of intratumor cytotoxic T cells that is not targeted by the current checkpoint inhibitors, but could be by anti-HLA-G/anti-ILT2 antibodies as novel immunotherapy in HLA-G⁺ tumors.

Keywords: CD4(+) CTLs; HLA-G; ILT2; Immune checkpoint; Renal carcinoma.