RNA seq of naïve CD8+ T cells expanded in different cytokines (along with our re-engineered cytokine) to find differences in t cell function, marturation and metabolic reprogramming.. RNA seq of naïve CD8+ T cells expanded in different cytokines (along with our re-engineered cytokine) to find differences in t cell function, marturation and metabolic reprogramming.

Cytokine therapy is limited by undesirable off-target side effects as well as terminal differentiation and exhaustion of chronically stimulated T cells. Here, we describe the signaling properties of a potentially unique cytokine by design, where T cell surface binding and signaling are separated between 2 different families of receptors. This fusion protein cytokine, called OMCPmutIL-2, bound with high affinity to the cytotoxic lymphocyte-defining immunoreceptor NKG2D but signaled through the common γ chain cytokine receptor. In addition to precise activation of cytotoxic T cells due to redirected binding, OMCPmutIL-2 resulted in superior activation of both human and murine CD8+ T cells by improving their survival and memory cell generation and decreasing exhaustion. This functional improvement was the direct result of altered signal transduction based on the reorganization of surface membrane lipid rafts that led to Janus kinase-3–mediated phosphorylation of the T cell receptor rather than STAT/AKT signaling intermediates. This potentially novel signaling pathway increased CD8+ T cell response to low affinity antigens, activated nuclear factor of activated T cells transcription factors, and promoted mitochondrial biogenesis. OMCPmutIL-2 thus outperformed other common γ chain cytokines as a catalyst for in vitro CD8+ T cell expansion and in vivo CD8+ T cell–based immunotherapy. Overall design: RNA-seq

细胞因子疗法（Cytokine therapy）受限于不良脱靶副作用，以及慢性刺激T细胞的终末分化与耗竭。本研究描述了一种经设计的潜在独特细胞因子的信号转导特性，该细胞因子将T细胞表面结合与信号转导功能分置于两类不同的受体家族中。这款被命名为OMCPmutIL-2的融合蛋白细胞因子，可与定义细胞毒性淋巴细胞的免疫受体NKG2D（NKG2D）以高亲和力结合，但通过共γ链细胞因子受体（common γ chain cytokine receptor）进行信号转导。除了通过重定向结合实现细胞毒性T细胞的精准激活外，OMCPmutIL-2还可通过提升人源与小鼠CD8+ T细胞的存活能力、促进记忆细胞生成并降低其耗竭，实现二者的更优激活。这一功能改善源于基于细胞膜脂筏（lipid rafts）重排的信号转导改变，该重排介导了T细胞受体的Janus激酶3（Janus kinase-3, JAK3）磷酸化，而非STAT/AKT信号中介分子。这一潜在新型信号通路可增强CD8+ T细胞对低亲和力抗原的应答，激活活化T细胞核因子转录因子（nuclear factor of activated T cells transcription factors），并促进线粒体生物发生（mitochondrial biogenesis）。因此，OMCPmutIL-2作为体外CD8+ T细胞扩增与体内基于CD8+ T细胞的免疫疗法的催化剂，其性能优于其他共γ链细胞因子。整体实验设计：RNA测序（RNA-seq）。