Centre for Precision Healthcare, Division of Medicine, University College London, London WC1E 6BT, United Kingdom.
Centre for Nature Inspired Engineering, University College London, London WC1E 6BT, United Kingdom.
ACS Appl Mater Interfaces. 2020 Oct 21;12(42):47355-47367. doi: 10.1021/acsami.0c16478. Epub 2020 Oct 7.
The recent rise of adoptive T cell therapy (ATCT) as a promising cancer immunotherapy has triggered increased interest in therapeutic T cell bioprocessing. T cell activation is a critical processing step and is known to be modulated by physical parameters, such as substrate stiffness. Nevertheless, relatively little is known about how biophysical factors regulate immune cells, such as T cells. Understanding how T cell activation is modulated by physical and biochemical cues may offer novel methods to control cell behavior for therapeutic cell processing. Inspired by T cell mechanosensitivity, we developed a multiwell, reusable, customizable, two-dimensional (2D) polyacrylamide (PA) hydrogel-integrated culture device to study the physicochemical stimulation of Jurkat T cells. Substrate stiffness and ligand density were tuned by concentrations of the hydrogel cross-linker and antibody in the coating solution, respectively. We cultured Jurkat T cells on 2D hydrogels of different stiffnesses that presented surface-immobilized stimulatory antibodies against CD3 and CD28 and demonstrated that Jurkat T cells stimulated by stiff hydrogels (50.6 ± 15.1 kPa) exhibited significantly higher interleukin-2 (IL-2) secretion, but lower proliferation, than those stimulated by softer hydrogels (7.1 ± 0.4 kPa). In addition, we found that increasing anti-CD3 concentration from 10 to 30 μg/mL led to a significant increase in IL-2 secretion from cells stimulated on 7.1 ± 0.4 and 9.3 ± 2.4 kPa gels. Simultaneous tuning of substrate stiffness and stimulatory ligand density showed that the two parameters synergize (two-way ANOVA interaction effect: < 0.001) to enhance IL-2 secretion. Our results demonstrate the importance of physical parameters in immune cell stimulation and highlight the potential of designing future immunostimulatory biomaterials that are mechanically tailored to balance stimulatory strength and downstream proliferative capacity of therapeutic T cells.
最近，过继性 T 细胞疗法（ATCT）作为一种有前途的癌症免疫疗法的兴起，引发了人们对治疗性 T 细胞生物加工的浓厚兴趣。T 细胞激活是一个关键的处理步骤，已知其可被物理参数（如基质硬度）调节。然而，人们对生物物理因素如何调节免疫细胞（如 T 细胞）知之甚少。了解 T 细胞激活如何受物理和生化线索调节，可能为控制治疗性细胞处理中的细胞行为提供新方法。受 T 细胞的机械敏感性启发，我们开发了一种多孔、可重复使用、可定制的二维（2D）聚丙烯酰胺（PA）水凝胶集成培养装置，以研究 Jurkat T 细胞的物理化学刺激。基质硬度和配体密度分别通过水凝胶交联剂的浓度和涂层溶液中的抗体浓度进行调节。我们在不同硬度的 2D 水凝胶上培养 Jurkat T 细胞，这些水凝胶表面固定有针对 CD3 和 CD28 的刺激抗体，并证明与较软水凝胶（7.1 ± 0.4 kPa）刺激的 Jurkat T 细胞相比，受硬性水凝胶（50.6 ± 15.1 kPa）刺激的 Jurkat T 细胞表现出更高的白细胞介素-2（IL-2）分泌，但增殖较低。此外，我们发现将抗 CD3 浓度从 10 μg/mL 增加到 30 μg/mL，会显著增加在 7.1 ± 0.4 和 9.3 ± 2.4 kPa 凝胶上受刺激的细胞中 IL-2 的分泌。同时调节基质硬度和刺激配体密度表明，这两个参数协同作用（双向方差分析交互作用：<0.001）以增强 IL-2 的分泌。我们的研究结果表明物理参数在免疫细胞刺激中的重要性，并强调了设计未来免疫刺激生物材料的潜力，这些材料可以根据力学特性进行定制，以平衡治疗性 T 细胞的刺激强度和下游增殖能力。
