The role of the intracellular complement system - the complosome - in Th1 biology

我们的目的是了解复合体在人类Th1反应的启动(产生干扰素-γ)和收缩(IL-10切换)以及CD8+CTL生物学中的作用。 答：我们在了解C3基因在T细胞和其他免疫细胞中的表达调控方面取得了重大进展。我们发现，整合素LFA-1(在免疫细胞进入组织的过程中参与)介导的信号是C3的主要诱导剂。因此，LFA-1缺乏的患者不能上调免疫细胞中的C3，从而缺乏正常的Th1、CTL和IL-1b产生巨噬细胞(Kolv和West，免疫学，2020)。 B.我们现在也已经能够确定CD46细胞内区域诱导人类CD4+T细胞中Th1相关基因和活动的确切分子机制。这是通过与细胞质中的某些代谢酶直接相互作用以及通过与细胞核中的特定转录因子相互作用而发生的(手稿正在准备中)。 C.在了解复合体如何共同诱导Th1细胞中的IL-10并启动其关闭程序方面，我们取得了同样的进展。复合体的这些新功能包括调节脂质和氨基酸使用的代谢酶的受控表达诱导(手稿正在准备中)。 D.为了试图以前所未有的深度理解朴素和记忆中的CD4+和CD8+T细胞激活的信号事件，我们对人类分类的T细胞进行了第一次随时间的比较scRNA-seq分析，并利用一种与U-Penn的张实验室合作的新算法分析了数据(手稿正在准备中)。

We aim at understanding the role of the complosome in both initiation (IFN-gamma production) and contraction (IL-10 switching) of human Th1 responses as well as CD8+ CTL biology. A. We have made major progress in understanding how C3 gene expression is regulated in T and other immune cells. We found that signals mediated by the integrin LFA-1 (engaged during diapedesis of immune cells into tissues) is a master inducer of C3. In consequence, patients with deficiency in LFA-1 cannot upregulate C3 in immune cells and, thus, lack normal Th1, CTL and IL-1b producing macrophages (Kolev and West, Immunity, 2020). B. We have now also been able to detangle the exact molecular mechanisms by which the intracellular domain of CD46 induces Th1-related genes and activities in human CD4+ T cells. This occurs via direct interaction with certain metabolic enzymes in the cytosol as well as via interaction with specific transcription factors in the nucleus (manuscripts in preparation). C. We have made equal progress in understanding how the complosome co-induces IL-10 in Th1 cells and initiates their shutdown program. These novel functions of the complosome involve the controlled expression induction of metabolic enzymes regulating lipid and amino acid usage (manuscripts in preparation). D. In an attempt to understand the signaling event underlying naive and memory CD4+ and CD8+ T cell activation in unprecedented depth, we have performed the first comparative scRNA-seq analysis of human sorted T cells over time and analyzed the data utilizing a novel algorithm integrating multiple unknown multivariate probability distributions in collaboration with the Zhang laboratory at U-Penn (manuscript in preparation).