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目的 研究IL 2基因修饰增强小鼠骨髓来源的树突状细胞 (DC)对肿瘤抗原提呈的功能和对MHCⅠ类限制性抗原多肽体内诱导的细胞毒性T淋巴细胞 (CTL)的激活作用及其相关的免疫机制。方法 用重组腺病毒介导IL 2基因修饰小鼠骨髓来源的DC ,ELISA法检测DC培养上清中IL 1 2和CTL上清中IFN γ的分泌水平 ,用流式细胞仪 (FACS)分析IL 2基因修饰对DC表面共刺激分子B7表达的调节和内吞卵清蛋白抗原八肽 (OVA)的作用 ,3H TdR掺入法检测DC对小鼠Lewis肺癌细胞株3LL肿瘤抗原的提呈能力 ;51 Cr释放法检测用 3LL细胞MHCⅠ类抗原多肽Mut1致敏IL 2基因修饰的DC对小鼠体内特异性CTL的诱导作用。结果 经IL 2基因修饰后 ,DC 48h能分泌高水平的IL 1 2(78.4± 6 .6)pg·(1× 1 0 6 细胞 ) - 1 ·ml- 1 ,DC表面的共刺激分子B7表达增加 ,DC内吞OVA多肽的作用也增强。经Mut1致敏后与同系 3LL细胞荷瘤的小鼠T淋巴细胞混合培养 ,3H TdR掺入量显著增高 ,用Mut1致敏IL 2基因修饰的DC免疫小鼠后 ,能在体内诱导出分泌高浓度IFN γ[(1 1 68.0± 58.4)pg/ml]的CTL活性。结论 IL 2基因修饰可活化DC抗原提呈的第二信号 ,增强DC对抗原多肽的捕获和提呈功能 ,经MHCⅠ类抗原多肽致敏后 ,在小鼠体内能更有效地诱导出CTL特异性抗肿瘤免疫应答。?
Objective To investigate the role of IL 2 gene modification in enhancing the antigen presenting function of mouse bone marrow-derived dendritic cells (DCs) and the activation of cytotoxic T lymphocytes (CTLs) induced in vivo by MHC class I-restricted antigen peptides Related immune mechanisms. Methods The bone marrow-derived DCs were modified with recombinant adenovirus mediated IL 2 gene. The secretion of IFN-γ in supernatant of IL-12 and CTL supernatants was detected by ELISA. The levels of IL-2 and IFN-γ in supernatants were determined by flow cytometry 2 gene modification on DC surface costimulatory molecule B7 expression and endocytic ovalbumin antigen octapeptide (OVA) role, 3H TdR incorporation assay of DC on mouse Lewis lung cancer cell line 3LL tumor antigen presentation ability; 51 Cr release method was used to detect the induction of specific CTLs in mice by DC sensitized with Mut2 sensitized by MHC class I antigen peptide of 3LL cells. Results After IL 2 gene modification, DCs secreted high levels of IL-12 (78.4 ± 6.6 pg · (1 × 10 6 cells) -1 · ml -1 at 48h after DC treatment, and the co-stimulatory molecule B7 expression on DC surface Increasing, the role of DC endocytosed OVA polypeptide also increased. After Mut1 sensitized with the same line 3LL tumor-bearing mouse T lymphocytes mixed culture, 3H TdR incorporation was significantly increased, with Mut1 sensitized IL2 gene-modified DC immunized mice induced in vivo high secretion Concentration of IFNγ [(1 1 68.0 ± 58.4) pg / ml] CTL activity. Conclusion IL 2 gene modification activates the second signal of DC antigen presentation and enhances the capture and presentation of antigen peptides by DCs. After being sensitized by MHC class I antigen peptide, CTL specificity can be induced more effectively in mice Anti-tumor immune response. ?