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目的 :探讨IL 12基因转染的树突状细胞 (DCs)能否体外加强细胞免疫反应。方法 :脂质体转染 (经过Flt 3L、SCF、GM CSF刺激 )增殖周期中的DCs前体细胞。加入GM CSF、IL 4和TNF α扩增转染细胞。在流式细胞仪上分析细胞表型及IL 12的表达。成熟DCs装载上分别能与HLA I类和HLA II类分子结合的多肽 ,观察IL 12基因转染DCs对特异性Th细胞发育及特异性CTL诱导和功能的影响。结果 :IL 12基因转染细胞在上述细胞因子作用下发育成为能有效表达IL 12并具有典型形态学与表型特征的DCs。在装载上CD4+T细胞表位HBcAg50 69后 ,能诱导这些细胞发育成为IFN γ产生型的Th1细胞。HLA A2 0 1亚型DCs在其同时装载CD4+T和CD8+T细胞相应的表位后 ,IL 12基因转染DCs所诱导产生的自身淋巴细胞的CTL效应增强。结论 :IL 12基因转染的DCs能增强对Th1细胞的刺激和CTL效应的诱导能力 ,提示它在肿瘤疫苗发展中的潜力
Objective: To investigate whether IL-12-transfected dendritic cells (DCs) can enhance cellular immune response in vitro. METHODS: Liposome-transfected DCs (pre-stimulated with Flt 3L, SCF, GM CSF) proliferate in DCs precursor cells. Transfected cells were expanded with GM CSF, IL4 and TNF [alpha]. Cell phenotype and IL 12 expression were analyzed on a flow cytometer. Matured DCs were loaded with peptides that could bind to HLA class I and HLA class II molecules, respectively. The effects of IL12 transfected DCs on specific Th cell development and specific CTL induction and function were observed. Results: IL 12 gene transfected cells developed DCs that could effectively express IL 12 and had typical morphological and phenotypic characteristics under the above cytokines. After loading with the CD4 + T cell epitope HBcAg5069, these cells were induced to develop into IFNγ-producing Th1 cells. The HLA-A2 0 subtype DCs enhanced the CTL effect of autologous lymphocytes induced by IL-12 gene-transfected DCs after they were simultaneously loaded with the corresponding epitopes of CD4 + T and CD8 + T cells. CONCLUSION: DCs transfected with IL-12 can enhance the induction of Th1 cells and the induction of CTL effects, suggesting its potential for tumor vaccine development