Sawaisorn Piamsiri, Tangchaikeeree Tienrat, Polpanich Duangporn, Midoeng Panuwat, Udomsangpetch Rachanee, Elaissari Abdelhamid, Jangpatarapongsa Kulachart
Center for Research and Innovation, Faculty of Medical Technology, Mahidol University Bangkok 10700 Thailand
National Nanotechnology Center, National Science and Technology Development Agency (NSTDA), Thailand Science Park Pathum Thani 12120 Thailand.
RSC Adv. 2018 Apr 17;8(26):14393-14400. doi: 10.1039/c8ra01468j.
γδ T cells play a significant role in protection against cancer. Purification of γδ T cells is needed for insight when studying their anti-cancer functionality and their utilization in adoptive cell therapy. To improve the purification of γδ T cells, in this work, a composite material based on magnetic nanoparticles was developed for purification of Vγ9Vδ2 T cells, the predominant subset of γδ T lymphocytes in human peripheral blood. The epoxy-functionalized magnetic poly(divinylbenzene--glycidyl methacrylate) particles (mPDGs) were bio-conjugated with anti-human Vδ2 antibody to provide specific recognition sites for T cell receptors of Vγ9Vδ2 T cells. Using fluorescence-activated cell sorting (FACS) analysis, separation of Vγ9Vδ2 T cells from peripheral blood mononuclear cells of healthy donors was confirmed with high purity [89.77% (range 87.00-91.80, = 3)]. More interestingly, the immobilized particles did not affect the viability of purified cells as high cell viability was indicated (>90%). By combining the properties of magnetic nanoparticles with specific antibodies, these immobilized particles were shown to be used as a cell-friendly purification tool of Vγ9Vδ2 T lymphocytes without any limits for the further use of cells. The purified Vγ9Vδ2 T cells using the antibody-immobilized epoxy-functionalized mPDGs could be used directly without a detachment step for further cultivation and expansion. This highlights the advantages of this method in allowing the study of cell function and further investigation of such rare T cell populations in immunotherapy.
γδ T细胞在抗癌保护中发挥着重要作用。在研究γδ T细胞的抗癌功能及其在过继性细胞疗法中的应用时，需要对其进行纯化以深入了解。为了改进γδ T细胞的纯化方法，在本研究中，开发了一种基于磁性纳米颗粒的复合材料，用于纯化人外周血中γδ T淋巴细胞的主要亚群Vγ9Vδ2 T细胞。环氧官能化的磁性聚（二乙烯基苯-甲基丙烯酸缩水甘油酯）颗粒（mPDGs）与抗人Vδ2抗体进行生物偶联，为Vγ9Vδ2 T细胞的T细胞受体提供特异性识别位点。通过荧光激活细胞分选（FACS）分析，证实从健康供体的外周血单核细胞中分离出的Vγ9Vδ2 T细胞具有高纯度[89.77%（范围87.00 - 91.80，n = 3）]。更有趣的是，固定化颗粒不会影响纯化细胞的活力，因为细胞活力较高（>90%）。通过将磁性纳米颗粒的特性与特异性抗体相结合，这些固定化颗粒被证明可作为一种对细胞友好的Vγ9Vδ2 T淋巴细胞纯化工具，且对细胞的进一步使用没有任何限制。使用抗体固定的环氧官能化mPDGs纯化的Vγ9Vδ2 T细胞无需进行解离步骤即可直接用于进一步培养和扩增。这突出了该方法在允许研究细胞功能以及进一步研究免疫疗法中此类稀有T细胞群体方面的优势。
