Advances in cell immunotherapy underscore the need for effective methods to produce large populations of effector T cells, driving growing interest in T-cell bioprocessing and immunoengineering. Research suggests that T cells demonstrate enhanced expansion and differentiation on soft matrices in contrast to rigid ones. Nevertheless, the influence of antibody conjugation chemistry on these processes remains largely unexplored. In this study, we examined the effect of antibody conjugation chemistry on T cell activation, expansion and differentiation using a soft and biocompatible polydimethylsiloxane (PDMS) platform. We rigorously evaluated three distinct immobilization methods, beginning with the use of amino-silane (PDMS-NH₂-Ab), followed by glutaraldehyde (PDMS-CHO-Ab) or succinic acid anhydride (PDMS-COOH-Ab) activation, in addition to the conventional physical adsorption (PDMS-Ab). By employing both stable amide bonds and reducible Schiff bases, antibody conjugation significantly enhanced antibody loading and density compared to physical adsorption. Furthermore, we discovered that the PDMS-COOH-Ab surface significantly promoted IL-2 secretion, CD69 expression, and T cell expansion compared to the other groups. Moreover, we observed that both PDMS-COOH-Ab and PDMS-NH₂-Ab surfaces exhibited a tendency to induce the differentiation of naïve CD4⁺ T cells into Th1 cells, whereas the PDMS-Ab surface elicited a Th2-biased immunological response. These findings highlight the importance of antibody conjugation chemistry in the design and development of T cell culture biomaterials. They also indicate that PDMS holds promise as a material for constructing culture platforms to modulate T cell activation, proliferation, and differentiation.

中文翻译：

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用于稳定抗体固定和 T 细胞培养的聚二甲基硅氧烷的表面化学修饰

细胞免疫疗法的进步凸显了对产生大量效应 T 细胞的有效方法的需求，从而推动了人们对 T 细胞生物加工和免疫工程日益增长的兴趣。研究表明，与刚性基质相比，T 细胞在软基质上表现出增强的扩增和分化能力。然而，抗体偶联化学对这些过程的影响在很大程度上仍未被探索。在这项研究中，我们使用柔软且生物相容的聚二甲基硅氧烷 (PDMS) 平台检查了抗体缀合化学对 T 细胞激活、扩增和分化的影响。我们严格评估了三种不同的固定方法，首先使用氨基硅烷 (PDMS-NH ₂ -Ab)，然后使用戊二醛 (PDMS-CHO-Ab) 或琥珀酸酐 (PDMS-COOH-Ab) 活化，此外与传统的物理吸附（PDMS-Ab）相比。通过采用稳定的酰胺键和可还原的希夫碱，与物理吸附相比，抗体缀合显着增强了抗体负载和密度。此外，我们发现与其他组相比，PDMS-COOH-Ab 表面显着促进 IL-2 分泌、CD69 表达和 T 细胞扩增。此外，我们观察到PDMS-COOH-Ab和PDMS-NH _{2 -Ab表面均表现出诱导幼稚CD4} ⁺ T细胞分化为Th1细胞的倾向，而PDMS-Ab表面引发了Th2偏向的免疫反应。这些发现强调了抗体偶联化学在 T 细胞培养生物材料设计和开发中的重要性。他们还表明，PDMS 有希望作为构建培养平台来调节 T 细胞活化、增殖和分化的材料。