Role of NFAT signaling in immune responses and protection against CMV

人类巨细胞病毒（HCMV）的控制取决于与宿主免疫反应的平衡。因此，CD8+ T细胞的过继免疫治疗足以缓解机会性HCMV感染，这种感染可能发生在同种异体造血（干细胞）移植（alloo - hct）中。在这些患者中，HCMV通过病毒细胞毒性直接引起疾病，但也加剧了共转移供体T细胞与宿主组织之间的免疫不相容性，从而加重了移植物抗宿主病（GvHD）。我们证明了激活t细胞家族核因子的两个成员（NFATc1和NFATc2）的信号传导不仅在免疫治疗后的GvHD中起部分冗余作用，而且在t细胞对小鼠CMV （MCMV）感染的反应中也起部分冗余作用。基于我们的研究结果，我们将利用MCMV潜伏感染小鼠的体内同种异体hct模型和HCMV感染并与人类NK和T细胞共培养的体外模型，研究NFAT信号在NK和T细胞对巨细胞病毒感染和GvHD的反应中的作用。我们将在人类NK或CD8+ T细胞中对NFATC1和/或NFATC2进行基因编辑，并测量它们在共培养细胞中限制HCMV复制的能力。因此，我们将确定NFAT在人类NK和CD8+ T细胞抗病毒活性中的相关性。体内活性将在MCMV模型中通过使用nfat消融的NK或T细胞的小鼠进行测试。我们将比较nfat缺陷小鼠和对照组小鼠NK记忆细胞的出现，引发的基因特征及其控制病毒复制的能力。此外，我们将在单倍体同源HCT和GvHD模型中研究这些细胞在潜伏感染小鼠中的作用。在这里，建立cmv特异性nfat缺陷T或NK细胞在细胞输注过程中应该处于的记忆状态以确保保护是很重要的。综上所述，我们将确定NFAT在临床相关环境中控制巨细胞病毒感染的作用和相关性。

Human cytomegalovirus (HCMV) control hinges on a balance with host immune responses. Hence, adoptive immunotherapy with CD8+ T cells is sufficient to provide relief against opportunistic HCMV infections, which might occur upon allogenic hematopoietic (stem) cell transplantation (allo-HCT). In these patients, HCMV causes disease directly via viral cytotoxicity, but also exacerbates immunological incompatibility between co-transferred donor T cells and host tissue, thus aggravating graft-versus-host disease (GvHD). We demonstrated that signaling by two members of the family nuclear factors of activated T-cells (NFATc1 and NFATc2) plays a partly redundant role not only in GvHD upon immunotherapy, but also in T-cell responses to mouse CMV (MCMV) infection. Building on our results, we will study NFAT signaling in NK- and T-cell responses to CMV infection and GvHD using in vivo models of allo-HCT in MCMV latently infected mice and in vitro models of HCMV infection and co-culture with human NK and T cells. We will gene-edit NFATC1 and/or NFATC2 in human NK or CD8+ T cells and measure their ability to limit HCMV replication in co-cultured cells. Thereby, we will define the relevance of NFAT in the antiviral activity of human NK and CD8+ T cells. In vivo activity will be tested in the MCMV model by using mice with NFAT-ablated NK or T cells. We will compare NFAT-deficient and control mice for the emergence of NK memory cells, the profile of genes elicited and their ability to control virus replication. Furthermore, we will study the effects of these cells in latently infected mice in models of haploidentical HCT and GvHD. Here, it is important to establish the memory status in which CMV-specific NFAT-deficient T or NK cells should be during cell infusion in order to ensure protection. Taken together, we will determine role and relevance of NFAT for controlling CMV infection in clinically relevant settings.