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. RNA-seq

细胞因子疗法（Cytokine therapy）常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。本研究报道一种经定制设计的潜在新型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过程分离，分别依赖两类不同的受体家族。这款命名为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，却通过共γ链细胞因子受体（common γ chain cytokine receptor）完成信号转导。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阳性T细胞的存活能力、促进记忆细胞生成，并降低其耗竭水平，实现二者的更优激活。该功能改善直接源于信号转导的改变：其通过重排细胞表面膜脂筏（lipid rafts），使T细胞受体（T cell receptor）被贾纳斯激酶3（Janus kinase-3）介导磷酸化，而非依赖STAT/AKT信号中介分子。这一潜在新型信号通路可增强CD8阳性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，并促进线粒体生物发生。因此，作为体外CD8阳性T细胞扩增与体内基于CD8阳性T细胞的免疫治疗的催化工具，OMCPmutIL-2的表现优于其他共γ链细胞因子。RNA测序（RNA-seq）