Waggoner Stephen, Cox Andrew, Canaday Laura, Katko Alexander, Feldman Harrison, Warrick Kathrynne, Tselikova Anastassia, Seelamneni Harsha, Roskin Krishna
Cincinnati Children's Hospital Medical Center.
UCLA.
Res Sq. 2024 Aug 30:rs.3.rs-4921081. doi: 10.21203/rs.3.rs-4921081/v1.
Natural killer (NK) cells suppress cellular and humoral immune responses via killing of T cells, resulting in diminished vaccine responses in mice and humans. Efforts to overcome this roadblock and achieve optimal immunity require an improved understanding of the molecular mediators facilitating NK cell-targeting of discrete subsets of CD4 T cells. We employed single-cell forensic victimology and CRISPR-Cas9 editing to delineate a transcriptional program uniquely responsible for the susceptibility of a subpopulation of CD4 T cells to perforin-dependent immunoregulation by NK cells. The unique vulnerability of these CD4 T cells relative to other subsets of CD4 T cells was not associated with a pattern of NK-cell-receptor ligand expression that would favor activation of NK cells. Instead, susceptible CD4 T cells were skewed toward follicular helper T cell (Tfh) differentiation and exhibited intermediate expression of Klf2 and a related suite of KLF2-target genes (e.g. S1pr1) involved in cell migration and spatial positioning. NK-cell dependent suppression of the subset of Tfh exhibiting intermediate expression of KLF2 and S1PR1 was confirmed with single-cell proteomics. CRISPR targeting of KLF2 in CD4 T cells prevented suppression by NK cells. Thus, KLF2 regulation of spatial positioning of T cells is a key determinant of NK-cell immunoregulatory function and a possible target for strategies to enhance vaccine efficacy.
自然杀伤（NK）细胞通过杀伤T细胞来抑制细胞免疫和体液免疫反应，导致小鼠和人类的疫苗反应减弱。要克服这一障碍并实现最佳免疫，需要更好地理解促进NK细胞靶向CD4 T细胞离散亚群的分子介质。我们采用单细胞法医学和CRISPR-Cas9编辑来描绘一个独特的转录程序，该程序负责CD4 T细胞亚群对NK细胞穿孔素依赖性免疫调节的敏感性。相对于其他CD4 T细胞亚群，这些CD4 T细胞的独特易感性与有利于NK细胞激活的NK细胞受体配体表达模式无关。相反，易感的CD4 T细胞倾向于滤泡辅助性T细胞（Tfh）分化，并表现出Klf2和一组相关的KLF2靶基因（如S1pr1）的中间表达，这些基因参与细胞迁移和空间定位。单细胞蛋白质组学证实了NK细胞对表现出KLF2和S1PR1中间表达的Tfh亚群的依赖性抑制。CRISPR靶向CD4 T细胞中的KLF2可防止NK细胞的抑制作用。因此，KLF2对T细胞空间定位的调节是NK细胞免疫调节功能的关键决定因素，也是增强疫苗效力策略的可能靶点。
