负载U266细胞抗原的树突状细胞诱导T细胞的抗骨髓瘤活性

Anti-myloma Activity of T cell Activated by Dentritic Cells Loading Antigen of U266 Cells

本研究探讨负载骨髓瘤U266细胞裂解物的树突状细胞(DC)瘤苗活化的T细胞对U266细胞的体外杀伤作用。用密度梯度离心法分离外周血单个核细胞,然后分别用含重组人粒细胞-巨噬细胞集落刺激因子(GM-CSF)、重组人白细胞介素-4(IL-4)的AIM-V无血清培养液和含胎牛血清的RPMI1640培养液体外诱导培养DC,用MTT法检测负载U266细胞全抗原的DC活化的T细胞对U266细胞的杀伤作用。结果表明:健康人外周血来源的单个核细胞用无血清培养液和含血清培养液诱导培养均可得到足够数量的DC,这两种培养液培养的DC在表型上无明显差异。分别用负载U266细胞全抗原的DC+IL-2、成熟DC+IL-2和单独应用IL-2刺激后的T细胞与U266细胞共培养,其对U266细胞的杀伤作用逐渐减弱,负载U266细胞全抗原的DC+IL-2刺激后的T细胞较成熟DC+IL-2刺激后的T细胞对U266细胞的杀伤作用明显,且T细胞对U266细胞的杀伤率随效靶比的增加而升高。结论:用GM-CSF、IL-4诱导培养健康人外周血单个核细胞可以得到不成熟的DC,体外负载U266细胞全抗原的DC可以刺激自体T细胞增殖并能杀伤U266细胞。

This study was aimed to investigate the effect of T cells activated by DCs loaded with whole antigens of U266 cells on the U266 cells survival in vitro. Peripheral blood mononuclear cells were isolated from healthy donor, and adherented on culture plate. Adherent cells were cultured in AIM-V serum-free medium or in RPMI 1640 medium contained 20% fetal bovine serum （FBS）, supplemented with granulocyte-macrophage colony stimulating factor （GM- CSF） and interleukin-4 （IL-4）. Methyl thiazolyl tetrazolium （MTT） assay was used to evaluate killing rate of U266 cells by T cells activated by DCs loaded with whole antigen of U266 cells. The results showed that DCs derived from peripheral blood mononuclear cells cultured by AIM-V serum-free medium or RPMI 1640 medium containing FBS had similar immunophenotype. T cells activated by DCs loaded with whole antigen of U266 cells or mature DCs might kill U266 cells in a dose-dependent manner. It is concluded, DCs derived from peripheral blood mononuclear cells of healthy donor and loaded with whole antigen of U266 cells can induce anti-myeloma response of T cells in vitro.