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目的 对比分析干扰素 γ(Interferon γ ,IFN γ)或脂多糖 (lipoplysaccharide ,LPS)刺激后的树突状细胞 (dendriticcell,DC)对肿瘤细胞杀伤活性的差异。方法 分离健康供者外周血单核细胞 ,用粒单细胞集落刺激因子和白介素 4诱导为DC。于培养液中加入LPS或IFN γ培养 12h ,作为LPS激活的DC(LPS DC)及IFN γ激活的DC(IFN DC)。用流式细胞仪检测DC表面共刺激分子的改变 ,以明确LPS或IFN γ对DC的不同刺激作用 ;同时 ,以恶性血液病细胞株HL 6 0、Jurkat及Daudi为靶细胞 ,用不同效靶比与DC共同培养 18h ,采用51Cr释放试验检测LPS DC及IFN DC抗肿瘤活性的差异。结果 ①LPS及IFN γ可不同程度的上调DC表面CD86、CD80、CD83及CD1a的表达 ,以LPS刺激组明显。②IFN γ和LPS可分别增强DC对HL6 0及Daudi的杀伤活性 ,在效靶比为 2 0∶1及 10∶1时杀伤率与未加刺激因子对照组 (medium DC)相比差异有显著意义 (P <0 0 5 )。相反 ,IFN γ DC对Daudi、LPS DC对HL 6 0无明显杀伤活性 ,但两者对Jurkat均具杀伤作用。结论 LPS及IFN γ激活的DC对肿瘤细胞的杀伤活性具有相对肿瘤特异性。
Objective To compare the difference in killing activity of dendritic cells (DCs) against tumor cells stimulated by interferon γ (IFN γ) or lipoplysaccharide (LPS). Methods Peripheral blood mononuclear cells were isolated from healthy donors and induced by granulocyte colony-stimulating factor and interleukin-4 as DC. LPS or IFNγ was added to the culture medium and cultured for 12 hours as LPS-activated DC (LPS DC) and IFNγ-activated DC (IFN DC). Flow cytometry was used to detect changes in DC surface co-stimulatory molecules to clarify the different stimulatory effects of LPS or IFN γ on DCs; at the same time, malignant hematological disease cell lines HL 60, Jurkat, and Daudi were used as target cells with different effect targets. Compared with DCs cultured for 18 hours, the 51Cr release assay was used to test the difference in antitumor activity of LPS DCs and IFN DCs. Results 1LPS and IFN γ could up-regulate the expression of CD86, CD80, CD83 and CD1a on DC surface, which was obviously observed in LPS-stimulated group. 2IFNγ and LPS enhanced the killing activity of DC on HL60 and Daudi, respectively, and the killing rate was significant at the effect ratio of 2:0∶1 and 10∶1, compared with the control group without medium (medium DC). (P < 0 0 5). On the contrary, IFN γ DC had no apparent killing activity against DHL and LPS DC against HL 60, but both had killing effect against Jurkat. Conclusion The killing activity of DCs activated by LPS and IFN γ is relatively tumor-specific.