Oncolytic vaccinia virus induces a novel phenotype of CD8⁺ effector T cells characterized by high ICOS expression

Characterization of the intratumoral immune status is important for developing immunotherapies and evaluating their antitumor effectiveness. CD8⁺ T cells are one of the most important cell types that directly and indirectly contribute to antitumor efficacy by releasing cytolytic molecules and inflammatory cytokines in the tumor microenvironment. Previously, we engineered a tumor-selective oncolytic vaccinia virus that encodes interleukin-7 (IL-7) and IL-12 and demonstrated its usefulness as an agent for in situ vaccination against tumors, with data showing that antitumor efficacy was reliant upon CD8⁺ T cells recruited by viral treatment. Here, we investigated the phenotypic changes in intratumoral CD8⁺ T cells caused by this oncolytic virus and found increased expression of inducible co-stimulator (ICOS) in PD-1^-CD8⁺ T cells. Unlike previously reported ICOS⁺CD8⁺ T cells, a subset of ICOS⁺PD-1^-CD8⁺ T cells showed effector function characterized by granzyme B expression. ICOS expression was induced by the backbone virus, which did not encode any immune transgenes and was independent of upregulation of the type I interferon pathway. Not only did we identify a novel effector cell subset characterized by ICOS expression, but our findings also shed light on a potential unknown aspect of the mechanism of oncolytic vaccinia virotherapy.