论文部分内容阅读
目的:探讨联合输注Flt3-L和GM-CSF真核表达质粒体内诱导小鼠DC的方法并检测其抗原提呈功能。方法:采用尾静脉注射法分别输注Flt3-L及GM-CSF质粒,15d后收获小鼠脾脏;采用流式细胞术检测小鼠脾细胞CD11c、CD11b、B220、CD8α和NK1.1等表面标志以鉴定DC的比例及亚型;将体内诱导DC与HBsAg共孵育,然后刺激HBsAg预免疫小鼠的脾细胞并检测其分泌IFN-的水平。结果:联合输注Flt3-L和GM-CSF质粒小鼠的脾细胞达到4×108个细胞/脾脏、其中CD11c+细胞占20%以上,CD11c+细胞中CD11b+、CD11b-、B220+、CD8+、NK1.1+细胞的比例分别达17%、10%、26%、16%、7%左右;体内诱导DC负载HB-sAg后刺激HBsAg特异性淋巴细胞分泌IFN-γ的水平明显高于HBsAg单独刺激组。结论:采用小鼠尾静脉注射技术输注Flt3-L及GM-CSF质粒能够诱导CD11c+ DC的产生并包含多种亚型,体内诱导DC具备有效的抗原提呈功能。
OBJECTIVE: To investigate the method of inducing mouse DC in vivo by co-infusion of Flt3-L and GM-CSF eukaryotic expression plasmids and to detect its antigen presenting function. Methods: Flt3-L and GM-CSF plasmids were respectively transfused by tail vein injection and spleen of mice were harvested 15 days later. Surface markers such as CD11c, CD11b, B220, CD8α and NK1.1 were detected by flow cytometry To identify the proportion of DC and subtype; in vivo induction of DC and HBsAg co-incubation, and then stimulate the pre-immunized mice HBsAg splenocytes and detect the secretion of IFN-. Results: The spleen cells of Flt3-L and GM-CSF plasmid-treated mice reached 4 × 108 cells / spleen, of which CD11c + cells accounted for more than 20%, CD11b +, CD11b-, B220 +, CD8 +, NK1.1 + Cells were 17%, 10%, 26%, 16% and 7% respectively. The level of IFN-γ secreted by HBsAg-specific lymphocytes stimulated by DC in vitro was significantly higher than that of HBsAg-stimulated group. CONCLUSION: The transfection of Flt3-L and GM-CSF plasmids by tail vein injection in mice can induce the production of CD11c + DC and include multiple subtypes. In vivo induction of DCs has an effective antigen presenting function.