Trans-signaling of the major pro- and anti-inflammatory cytokines Interleukin (IL)-6 and IL-11 has the unique feature to virtually activate all cells of the body and is critically involved in chronic inflammation and regeneration. Hyper-IL-6 and Hyper-IL-11 are single chain designer trans-signaling cytokines, in which the cytokine and soluble receptor units are trapped in one complex via a flexible peptide linker. Albeit, Hyper-cytokines are essential tools to study trans-signaling in vitro and in vivo, the superior potency of these designer cytokines are accompanied by undesirable stress responses. To enable tailor-made generation of Hyper-cytokines, we developed inactive split-cytokine-precursors adapted for posttranslational reassembly by split-intein mediated protein trans-splicing (PTS). We identified cutting sites within IL-6 (E¹³⁴/S¹³⁵) and IL-11 (G¹¹⁶/S¹¹⁷) and obtained inactive split-Hyper-IL-6 and split-Hyper-IL-11 cytokine precursors. After fusion with split-inteins, PTS resulted in reconstitution of active Hyper-cytokines, which were efficiently secreted from transfected cells. Our strategy comprises the development of a background-free cytokine signaling system from reversibly inactivated precursor cytokines.

中文翻译：

---

拆分²蛋白连接从无活性的前体产生活性的IL-6型超细胞因子

主要促炎细胞因子和抗炎细胞因子的转信号白介素（IL）-6和IL-11具有独特的功能，可以虚拟激活人体的所有细胞，并且与慢性炎症和再生密切相关。Hyper-IL-6和Hyper-IL-11是单链设计者的反信号细胞因子，其中的细胞因子和可溶性受体单元通过一种柔性肽接头被捕获在一种复合物中。尽管，超细胞因子是研究反式信号传导的重要工具在体外和体内这些设计细胞因子的强大效能伴随着不良的应激反应。为了实现量身定制的超细胞因子生成，我们开发了无活性的分裂细胞因子前体，适用于通过分裂内含蛋白介导的蛋白质反式剪接（PTS）进行翻译后重组。我们确定了IL-6（E ¹³⁴ / S ¹³⁵）和IL-11（G ¹¹⁶ / S ¹¹⁷）内的切割位点，并获得了非活性的Hyper-IL-6和Split-Hyper-IL-11细胞因子前体。与分裂内含蛋白融合后，PTS导致活性超细胞因子的重建，该因子从转染的细胞中有效分泌。我们的策略包括从可逆灭活的前体细胞因子开发无背景的细胞因子信号系统。