Fellermeier-Kopf Sina, Gieseke Friederike, Sahin Ugur, Müller Dafne, Pfizenmaier Klaus, Kontermann Roland E
Institute of Cell Biology and Immunology, University of Stuttgart, Stuttgart, Germany.
BioNTech RNA Pharmaceuticals GmbH, Mainz, Germany.
Oncoimmunology. 2018 Aug 1;7(9):e1471442. doi: 10.1080/2162402X.2018.1471442. eCollection 2018.
Co-stimulatory signals induced by ligands of the tumor necrosis factor superfamily (TNFSF) play a central role in T cell activation and have emerged as a promising strategy in cancer immunotherapy. Here, we established a novel class of bifunctional co-stimulatory fusion proteins with the aim to boost T cell activation at the level of T cell - antigen-presenting cell (APC) interaction. These novel dual-acting cytokine fusion proteins were created by connecting two different homotrimeric TNFSF ligands to form homotrimeric bifunctional molecules (Duokines) or by connecting single-chain derivatives of two different homotrimeric TNFSF with a single, flexible linker (single-chain Duokines, scDuokines). By linking the TNFSF ligands 4-1BBL, OX40L and CD27L in all possible combinations, cis-acting Duokines were generated that act on the same or adjacent T cells, while combining CD40L with 4-1BBL, OX40L and CD27L resulted in trans-acting Duokines acting simultaneously on APCs and T cells. , co-stimulation of T cells was seen for cis- and trans-acting Duokines and scDuokines in an antigen-independent as well as antigen-specific setting. Trans-acting molecules furthermore activated B cells, which represent a subclass of APCs. In a pilot experiment using the syngeneic B16-FAP mouse tumor model scDuokines displayed antitumoral activity in vivo in combination with a primary T cell-activating bispecific antibody, evident from reduced number of lung metastasis compared to the antibody-only treated group. Our data show that the bifunctional, co-stimulatory duokines are capable to enhance T cell-mediated anti-tumor immune responses, suggesting that they can serve as a new class of immuno-stimulatory molecules for use in cancer immunotherapy strategies.
肿瘤坏死因子超家族（TNFSF）配体诱导的共刺激信号在T细胞活化中起核心作用，并已成为癌症免疫治疗中有前景的策略。在此，我们建立了一类新型双功能共刺激融合蛋白，旨在在T细胞与抗原呈递细胞（APC）相互作用水平上增强T细胞活化。这些新型双作用细胞因子融合蛋白通过连接两种不同的同三聚体TNFSF配体形成同三聚体双功能分子（双激动剂），或通过用单个柔性接头连接两种不同同三聚体TNFSF的单链衍生物（单链双激动剂，sc双激动剂）来产生。通过以所有可能的组合连接TNFSF配体4-1BBL、OX40L和CD27L，产生了作用于相同或相邻T细胞的顺式作用双激动剂，而将CD40L与4-1BBL（ OX40L和CD27L）组合则产生了同时作用于APC和T细胞的反式作用双激动剂。在抗原非依赖性以及抗原特异性环境中，顺式和反式作用双激动剂和sc双激动剂均可见对T细胞的共刺激作用。此外，反式作用分子激活了作为APC亚类的B细胞。在使用同基因B16-FAP小鼠肿瘤模型的初步实验中，sc双激动剂与原发性T细胞激活双特异性抗体联合在体内显示出抗肿瘤活性，与仅用抗体治疗的组相比，肺转移数量减少证明了这一点。我们的数据表明，双功能共刺激双激动剂能够增强T细胞介导的抗肿瘤免疫反应，表明它们可作为一类新的免疫刺激分子用于癌症免疫治疗策略。
