锚定修饰磁珠的SA-hIL2/7/21体外扩增人外周血T细胞的实验研究

The expansion of human peripheral blood T cells by SA-hIL2/7/21 anchored beads in vitro

目的为探究一种新的T细胞体外扩增方式,探索并比较了SA-hIL2、SA-hIL7、SA-hIL21 3种细胞因子在游离与锚定修饰磁珠2种情况下,对T细胞的增殖以及表型特征的影响。方法利用本实验室大肠杆菌原核表达系统制备的SA-hIL2/7/21融合蛋白锚定修饰磁珠后体外扩增T细胞,随后CFSE标记流式细胞术分别测定细胞的增殖程度以及初始记忆T细胞表面标志的表达。结果锚定修饰磁珠的SA-hIL2、SA-hIL7、SA-hIL21促增殖效果显著优于游离的SA-hIL2、SA-hIL7、SA-hIL21,并且前者可促进CD8^+CD45RO^+以及CD62L-CCR7-(TEM)表达,但降低CD8^+CD45RO-和CD62L^+CCR7^+(TCM)的表达。结论与游离组扩增T细胞相比,锚定修饰磁珠组一方面促进T细胞增殖的能力更强;另一方面可使T细胞表型由TCM向TEM转变。

This study was designed to explore a new method for in vitro expansion of T cells, providing a reference for adoptive immunotherapy to generate sufficient amounts of T cells for transmission. The recombinant SA-hIL2/7/21 fusion protein prepared in the E. coli prokaryotic expression system in our laboratory was used, and the proteins were then anchored to the biotinylated magnetic beads by the specific binding of biotin to streptavidin. T cells were isolated from peripheral blood of healthy volunteers, and then cultured with free proteins and proteins anchored to magnetic beads, respectively. Then CFSE-labeled flow cytometry was used to determine the degree of cell proliferation and the expression of initial memory T cell markers. Data showed that the SA-hIL2, SA-hIL7, and SA-hIL21 anchorage-modified magnetic beads significantly proliferated better than free SA-hIL2, SA-hIL7 and SA-hIL21; the anchored group promoted the expression of CD8~＋CD45 RO~＋and CD62 L-CCR7-（TEM） but reduced expression of CD8~＋CD45 RO-and CD62 L~＋CCR7~＋（TCM）. Finally, it was concluded that the anchorage-modified magnetic bead group not only has a stronger ability to promote T cell proliferation but also change the T cell phenotype from TCM to TEM.