Maghsoodi Nafiseh, Zareinejad Mohammadrasul, Ghaderi Abbas, Mahmoudi Maymand Elham, Irajie Cambyz, Ramezani Amin
Student Research Committee, Shiraz University of Medical Sciences, Shiraz, Iran; Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Iran; Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
Cytokine. 2025 Jan;185:156822. doi: 10.1016/j.cyto.2024.156822. Epub 2024 Dec 3.
To overcome the limitations of IL-15 and to improve the efficacy of IL-15 in immunotherapy, several strategies have been introduced.
The objective of this study was to generate and evaluate a novel anti-CD8/IL-15 (N72D)/Sushi fusion protein with the potential to target CD8 T cells and enhance functionality of CD8 T cells against tumor cells.
In this connection, a novel fusokine that contains IL-15(N72D), a Sushi domain, and anti-CD8 single-chain fragment variable (scFv) was designed. The size accuracy and binding potency of the isolated protein were assessed using western blotting and indirect surface staining. Following purification, the potential function of the anti-CD8/IL-15(N72D)/Sushi fusion protein in the induction of proliferation and cytotoxicity of CD8 T cells was evaluated.
In-silico analysis revealed that fusokine is structurally stable, correctly folded and can interact with the CD8 co-receptor. Both fusokine and IL-15(N72D)/Sushi were produced in CHO-S cell line with a final concentration of 18.43 mg/l and 12.64 mg/l respectively. Fusokine bound to 97.6 % of CD8 T cells and significantly induced T cell proliferation and cytotoxic potential in peripheral blood mononuclear cells (PBMCs) in a time dependent manner. Compared to both the control and the IL-15 (N72D)/sushi treated groups, fusokine showed superior potential in CD8 T cell functionality.
Anti-CD8/IL-15(N72D)/Sushi has the ability to effectively target CD8 T cells, promote lymphocyte proliferation and induce cytotoxicity against tumor cells. Due to its promising properties, it could be considered as a new potential immunotherapy approach.
为克服白细胞介素 - 15（IL - 15）的局限性并提高其在免疫治疗中的疗效，已引入多种策略。
本研究的目的是生成并评估一种新型抗CD8/IL - 15（N72D）/寿司融合蛋白，该蛋白具有靶向CD8 T细胞并增强CD8 T细胞对肿瘤细胞功能的潜力。
为此，设计了一种新型融合细胞因子，其包含IL - 15（N72D）、一个寿司结构域和抗CD8单链可变片段（scFv）。使用蛋白质印迹法和间接表面染色评估分离蛋白的大小准确性和结合能力。纯化后，评估抗CD8/IL - 15（N72D）/寿司融合蛋白在诱导CD8 T细胞增殖和细胞毒性方面的潜在功能。
计算机模拟分析表明，融合细胞因子结构稳定、折叠正确且能与CD8共受体相互作用。融合细胞因子和IL - 15（N72D）/寿司均在CHO - S细胞系中产生，最终浓度分别为18.43 mg/l和12.64 mg/l。融合细胞因子与97.6%的CD8 T细胞结合，并以时间依赖性方式显著诱导外周血单核细胞（PBMCs）中T细胞增殖和细胞毒性潜力。与对照组和IL - 15（N72D）/寿司处理组相比，融合细胞因子在CD8 T细胞功能方面表现出更优的潜力。
抗CD8/IL - 15（N72D）/寿司具有有效靶向CD8 T细胞、促进淋巴细胞增殖并诱导对肿瘤细胞的细胞毒性的能力。由于其具有良好的特性，可被视为一种新的潜在免疫治疗方法。
