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目的:采用细胞因子诱导的杀伤细胞(cytokine induced killer cell,CIK cell)培养基(含低浓度IL-2、IFN-γ、IL-12和anti-CD3)和NK培养基(含高浓度IL-2和anti-CD3)体外刺激诱导CIK,分析CIK细胞的扩增、表型和细胞因子分泌情况。方法:健康自愿者来源的外周血单个核细胞(peripheral blood mononuclear cell,PBMC),分别加入CIK培养基和NK培养基进行诱导培养,24 h后,换用含IL-2的1%自体血浆培养基继续诱导培养2周。在培养的第6天和第10天检测培养上清中IFN-γ的分泌情况,在培养的第10天以流式细胞术检测两组培养细胞表面CD3、CD4、CD8、CD16、CD56的表达情况。结果:CIK培养基和NK培养基均可使培养细胞在第7天时开始明显扩增,第10天时可扩增至200倍。其中,CIK培养基获得的细胞扩增倍数明显高于NK培养基,其IFN-γ分泌明显高于NK培养基组[(724.7±434.8)vs(108.9±60.6)pg/ml,P<0.01];而以NK培养基培养的细胞,CD3-CD16+CD56+细胞比例高于CIK培养基[(6.27±3.33)%vs(2.88±1.88)%,P<0.05]。结论:不同成分的CIK培养基和NK培养基均可促进CIK细胞的扩增和成熟,但呈现不同的生物学特征。
OBJECTIVE: To investigate the effect of IL-2 on IL-2, IFN-γ, IL-12 and anti-CD3 production by cytokine induced killer cell (CIK cell) 2 and anti-CD3) in vitro induced CIK, CIK cell proliferation, phenotype and cytokine secretion. Methods: Peripheral blood mononuclear cells (PBMCs) derived from healthy volunteers were cultured in CIK medium and NK medium respectively. After 24 h, they were cultured in 1% autologous plasma containing IL-2 Continue to induce induction culture for 2 weeks. The secretion of IFN-γ in the culture supernatant was detected on the 6th day and the 10th day of culture, and the expression of CD3, CD4, CD8, CD16 and CD56 on the cultured cells were detected by flow cytometry on the 10th day of culture Happening. Results: Both CIK medium and NK medium could significantly proliferate cultured cells on the 7th day and 200 times on the 10th day. Among them, CIK medium obtained cell proliferation fold was significantly higher than the NK medium, the IFN-γ secretion was significantly higher than the NK medium group (724.7 ± 434.8 vs 108.9 ± 60.6 pg / ml, P 0.01] ; While the proportion of CD3-CD16 + CD56 + cells cultured in NK medium was higher than that of CIK medium [(6.27 ± 3.33)% vs (2.88 ± 1.88)%, P <0.05]. CONCLUSION: CIK and NK cultures with different components all promote CIK cell proliferation and maturation, but show different biological characteristics.