NFAT2 Regulates Generation of Innate-Like CD8⁺ T Lymphocytes and CD8⁺ T Lymphocytes Responses

Nuclear factor of activated T cells (NFAT) 2 null mutant mice die in utero of cardiac failure, precluding analysis of the role of NFAT2 in lymphocyte responses. Only the NFAT2^-/-/Rag-1^-/- chimeric mice model gave insight into the role of NFAT2 transcription factor in T lymphocyte development, activation, and differentiation. As reports are mainly focused on the role of NFAT2 in CD4⁺ T lymphocytes activation and differentiation, we decided to investigate NFAT2's impact on CD8⁺ T lymphocyte responses. We report that NFAT2 is phosphorylated and inactive in the cytoplasm of naive CD8⁺ T cells, and upon TCR stimulation, it is dephosphorylated and translocated into the nucleus. To study the role of NFAT2 in CD8⁺ T responses, we employed NFAT2^fl/flCD4-Cre mice with NFAT2 deletion specifically in T cells. Interestingly, the absence of NFAT2 in T cells resulted in increased percentage of non-conventional innate-like CD8⁺ T cells. These cells were CD122⁺, rapid producer of interferon gamma (IFN-γ) and had characteristics of conventional memory CD8⁺ T cells. We also observed an expansion of PLZF⁺ expressing CD3⁺ thymocyte population in the absence of NFAT2 and increased IL-4 production. Furthermore, we found that CD8⁺ T lymphocytes deficient in NFAT2 had reduced activation, proliferation, and IFN-γ and IL-2 production at suboptimal TCR strength. NFAT2 absence did not significantly influence differentiation of CD8⁺ T cells into cytotoxic effector cells but reduced their IFN-γ production. This work documents NFAT2 as a negative regulator of innate-like CD8⁺ T cells development. NFAT2 is required for complete CD8⁺ T cell responses at suboptimal TCR stimulation and regulates IFN-γ production by cytotoxic CD8⁺ T cells in vitro.