【摘要】 目的　探讨转染干扰素γ诱导蛋白 10 (IP 10 )基因构建的加载树突状细胞 (DC)瘤苗 ,对特异性细胞毒T淋巴细胞 (CTL)的诱导作用。　方法　将RM 1细胞的裂解产物作为肿瘤抗原加载小鼠骨髓来源的DC(Tuly DC) ,将IP 10DNA插入真核表达质粒 ,通过脂质体法将IP 10基因转染至Tuly DC ,构建DC瘤苗 ;RT PCR法检测IP 10基因转染成功 ,趋化实验检测对淋巴细胞的趋化作用 ,混合淋巴细胞反应 (MLR)检测瘤苗刺激T细胞的增殖能力 ,MTT法检测瘤苗诱导的特异性CTL的杀伤活性。　结果　转染DC的IP 10表达明显增强 ,其上清对淋巴细胞有较强的趋化作用 ;DC瘤苗刺激T细胞增殖能力增强 ,每分钟闪烁计数值分别为 18913.0 9± 2 735 .33(1∶10 )和 17736 .6 7± 2 5 31.70 (1∶2 0 ) ,与各对照组相比差异有显著性意义 (P <0 .0 1) ;DC瘤苗诱导的CTL对RM 1细胞的特异性杀伤率分别为 (37.95± 5 .2 6 ) % (效靶比 2 0∶1)和 (43.87± 7.6 3) % (效靶比 4 0∶1) ,均高于各对照组 ,差异有显著性意义 (P <0 .0 1)。　结论　所构建的前列腺癌DC瘤苗能有效提高其抗原提呈功能和特异性CTL的诱导。更多还原

【Abstract】 Objective To investigate the effect of murine dentritic cells vaccine modified with IFN-γ inducible protein-10 gene on CTL induction. Methods DC was propagated from bone marrow (BM) of mice and pulsed with mouse prostate cancer cell line RM-1’s whole lysate (Tuly-DC).IP-10 DNA fragments were inserted into pcDNA3.1(+) vector to construct recombinant plasmid IP-10/pcDNA3.1.Tuly-DC was transfected with IP-10/pcDNA3.1 by DOTAP liposome.Reversal transcript-polymerase chain reaction (RT-PCR) was used to evaluate gene transfer efficiency and chemotaxis assay was used to estimate the tansfected DC’s chemotactic activity on T cells.Antitumor activity of the DC vaccine was studied in vitro by using Mixed leukocyte reaction (MLR) and cytotoxic assay (MTT assay). Results The IP-10 plasmid vector was successfully transfected into DC,which was confirmed by RT-PCR.The DC tranfected with IP-10 gene was capable of synthesizing and secreting IP-10 chemokine,which could increase the preferential chemotaxis of DC to T cells.MLR showed that the DC pulsed with whole tumor lysate and modified with IP-10 gene (IP-10/ Tuly-DC) could induce T cell proliferation significantly compared with other groups (P<0.01), the CPM were 18 913.09±2 735.33 (1 ∶10) and 17 736.67±2 531.70 (1 ∶20). The highest CTL activity against RM-1 cells was induced in mice immunized with IP-10/ Tuly-DC compared with that in mice of other groups (P<0.01), the specific lysis were (37.95±5.26)%(20 ∶1) and (43.87±7.63)%(40 ∶1). Conclusions DC vaccine pulsed with whole tumor lysate and modified with IP-10 gene can induce a specific and effective immune response against prostate cancer cells.更多还原