Research Group in Immunobiology and Therapeutic Sciences, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkoknoi, Bangkok, Thailand.
Biomedical Research Incubator Unit, Research Group and Research Network Division, Research Department, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkoknoi, Bangkok, Thailand.
Scand J Immunol. 2019 Nov;90(5):e12808. doi: 10.1111/sji.12808. Epub 2019 Aug 22.
CD4 T cell immunotherapy has potential for treatment in HIV-infected patients. A large number of expanded CD4 T cells and confirmation of functional-related phenotypes are required for ensuring the successful outcomes of treatment. Freshly isolated CD4 T cells from healthy donors were activated with anti-CD3/28-coated magnetic beads at different bead-to-cell ratios and cultured in the absence and presence of IL-2 supplementation for 3 weeks. Fold expansion, cell viability, growth kinetic and lymphocyte subset identities were determined. Data demonstrated that a 1:1 bead-to-cell ratio rendered the highest expansion of 1044-fold with 88% viability and 99.5% purity followed by the 2:1 and 0.5:1 ratios. No significant difference in proliferation and phenotypes was found between non-IL-2 and IL-2 supplementation groups. Several specific surface molecule expressions of the expanded cells including chemokine receptors, adhesion molecules, co-stimulatory molecules, activation molecules, maturation markers, cytokine receptors and other molecules were altered when compared to the unexpanded cells. This optimized expansion protocol using the 1:1 bead-to-cell ratio of anti-CD3/28-coated magnetic beads and culture condition without IL-2 supplementation provided the satisfactory yield with good reproducibility. Specific surface molecule expressions of the expanded cells presented potential roles in proliferation, differentiation, homeostasis, apoptosis and organ homing.
CD4 T 细胞免疫疗法在治疗 HIV 感染患者方面具有潜力。为了确保治疗的成功，需要大量扩增的 CD4 T 细胞并确认其功能相关表型。从健康供体中分离出新鲜的 CD4 T 细胞，用抗 CD3/28 包被的磁珠在不同的磁珠与细胞比例下激活，并在无和有 IL-2 补充的情况下培养 3 周。测定细胞扩增倍数、细胞活力、生长动力学和淋巴细胞亚群身份。数据表明，磁珠与细胞的比例为 1:1 时，细胞活力为 88%，纯度为 99.5%，扩增倍数最高可达 1044 倍，其次是磁珠与细胞的比例为 2:1 和 0.5:1。无 IL-2 和有 IL-2 补充的两组之间的增殖和表型没有显著差异。与未扩增细胞相比，扩增细胞的几种表面分子表达发生了变化，包括趋化因子受体、粘附分子、共刺激分子、激活分子、成熟标志物、细胞因子受体和其他分子。使用抗 CD3/28 包被的磁珠的 1:1 磁珠与细胞比例和无 IL-2 补充的培养条件优化的扩增方案提供了令人满意的产量和良好的可重复性。扩增细胞的表面分子表达在增殖、分化、稳态、凋亡和器官归巢方面具有潜在作用。
