肿瘤患者偏移的T细胞TCRBV表达谱在自体CIK细胞中的恢复

Recovery of biased TCRBV profile of tumor patients after in vitro autologous CIK cell culture

目的研究肿瘤患者自体PBMC经体外诱导培养的CIK细胞群TCRBV亚家族表达的变化。方法Ficoll密度梯度离心法分类人体外周血单核细胞（PBMC）,经IFN-γ、OKT-3以及IL-2诱导培养,Trizol试剂提取总RNA,RT-PCR半定量分析24个TCRBV亚家族表达情况,流式分选平台分选CD3＋CD8＋细胞和CD3＋CD56＋细胞亚群。结果诱导培养后的CIK中,分化的CD3＋CD56＋细胞亚群比例占12.6%~41.7%。PBMC中弱表达或缺失的TCRBV亚家族,在CIK中上调或恢复表达。CIK细胞群中,CD3＋CD8＋细胞和CD3＋CD56＋细胞亚群克隆组成无明显差异。结论肿瘤患者自体PBMC体外诱导培养CIK细胞后,部分表达偏移的TCRBV亚家族能恢复表达,表明部分受损或被抑制的T细胞克隆在CIK培养中被激活并增殖。

Objective To investigate the variation of TCRBV subfamily between autologous PBMC from tumor patients and in vitro cultured CIK. Methods PBMC was separated by Ficoll gradient centrifugation,and cultured with IFN-γ,OKT-3 and IL-2. Total RNA was separated by Trizol,and RT-PCR was performed to analyze the expression of TCRBV 24 subfamilies. FACS was used to sort CD3＋CD8＋cells and CD3＋CD56＋cells. Results The ratio of differentiated CD3＋CD56＋cells in cultured CIK accounted for 12. 6%-41. 7%. The expression of TCRBV subfamilies decreased or deficient in PBMC was up-regulated in CIK.There was no significant difference in clonal combination between CD3＋CD8＋cells and CD3＋CD56＋cells. Conclusion The biased TCRBV profile can be partly recovered during in vitro CIK culture,suggesting that some T cells impaired or inhibited from tumor patient may be activated and enter replication cycle.