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目的 观察mIL 12基因转染的小鼠树突状细胞(DC)功能的改变。方法 分离小鼠骨髓单个核细胞与rmGM CSF和rmIL 4培养 1周 ,对培养的细胞进行形态学观察 ,FACS检测细胞表面DEC2 0 5、CD86表达。以mIL 12重组腺病毒转染培养的DC ,ELISA测定培养上清中mIL 12的水平 ,混合淋巴细胞反应检测转染细胞的功能。结果 培养 1周后 ,得到具有典型DC形态的细胞 ,以mIL 12重组腺病毒为载体转染DC ,培养上清中可以检测到较高水平的mIL 12 ;与对照组相比转染细胞能明显地刺激混合林巴细胞反应。结论 小鼠骨髓单个核细胞经GM CSF和IL 4培养 1周 ,生成大量DC ,mIL 12重组腺病毒感染培养的DC对T细胞刺激显著增强
Objective To observe the function of mouse dendritic cells (DCs) transfected with mIL 12 gene. Methods Mice bone marrow mononuclear cells were cultured with rmGM CSF and rmIL 4 for 1 week. The cultured cells were observed morphologically. The expression of DEC2 05 and CD86 was detected by FACS. Cultured DCs were transfected with the recombinant adenovirus of mIL 12, the level of mIL 12 in the culture supernatant was measured by ELISA, and the function of the transfected cells was examined by mixed lymphocyte reaction. Results After culturing for 1 week, cells with typical DC morphology were obtained. DCs were transfected with mIL 12 recombinant adenovirus and higher levels of mIL 12 were detected in the culture supernatant. Compared with the control group, transfected cells could be significantly To stimulate mixed lymbal cell responses. Conclusion Mice bone marrow mononuclear cells were cultured with GM CSF and IL 4 for 1 week to generate a large amount of DCs. DCs infected with mIL 12 recombinant adenovirus significantly stimulated T cell stimulation