瘦素基因佐剂对小鼠抗原提呈细胞表型的影响  

The effect of leptin genetic adjuvant on the phenotypes of mouse antigen-presenting cells

作　　者：郭贺[1] 高琦[1] 徐丽慧[2] 欧阳东云[1] 刘春永[1] 何贤辉[1] 

机构地区：[1]暨南大学生命科学技术学院组织移植与免疫实验中心,广东广州510632 [2]暨南大学生命科学技术学院生物工程研究所,广东广州510632

出　　处：《暨南大学学报（自然科学与医学版）》2010年第2期125-129,共5页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：国家自然科学基金项目(30572199;30371651)

摘　　要：目的:研究瘦素(Leptin)基因佐剂在体内对抗原提呈细胞的影响。方法:根据小鼠瘦素基因设计引物,采用RT-PCR方法获得瘦素cDNA,与pVAX1连接构建瘦素真核表达载体作为基因佐剂,制备无内毒素重组质粒,转染B16细胞鉴定瘦素的表达,并采用肌注方式分析瘦素基因佐剂在体内对小鼠抗原提呈细胞表型的影响。结果:克隆得到的瘦素编码区全长序列,经DNA测序后证明与已报道序列相同,成功构建小鼠瘦素的真核表达载体,可在B16细胞表达;瘦素基因佐剂在体内对CD3-CD19-CD11c+细胞的MHC II和CD83的表达无明显影响,但能显著上调CD3-CD19-CD11c-免疫细胞表面MHC II类分子的表达。结论:成功构建小鼠瘦素基因佐剂,体内试验可促进免疫细胞上调MHC II类分子,提示瘦素基因佐剂有可能通过增强免疫细胞尤其是抗原提呈细胞的活化而促进免疫应答。Aim：To investigate the effect of leptin genetic adjuvant on the antigen-presenting cells in vivo. Methods： The cDNA for leptin was amplified by RT-PCR using a pair of primers designed according to mouse leptin gene and then inserted into plasmid pVAX1 as a genetic adjuvant.The recombinant plasmid that had no endotoxin was prepared and its expression in B16 cells was verified by RT-PCR.The in vivo effect of leptin genetic adjuvant on the phenotypes of mouse antigen-presenting cells was examined by intramuscular injection.Results： The results showed that the cloned cDNA encoding leptin was identical to the published sequence after DNA sequence analysis.The eukaryotic expression vector encoding leptin was constructed and successfully expressed in B16 cells.In vivo assay indicated that the expression of MHC II and CD83 on CD3^-CD19^-CD11c^＋ immune cells was not affected by the leptin genetic adjuvant,but the expression of MHC II on CD3^-CD19^-CD11c^-cells was significantly up-regulated.Conclusion：It was evident that the leptin genetic adjuvant could up-regulate the expression of MHC II on immune cells,suggesting that the immune response may be enhanced by leptin genetic adjuvant through the activation of immune cells,especially antigen-presenting cells.

关 键 词：瘦素 基因佐剂 真核表达 抗原提呈细胞 

分 类 号：R392.9[医药卫生—免疫学]