摘要
目的观察Rab4突变体在树突状细胞(DC)内的定位,研究其对DC内源抗原递呈的影响。方法构建Rab4突变体的慢病毒表达质粒,以DNA-磷酸钙共沉淀法制备慢病毒,病毒感染DC-OVA细胞并使用流式细胞术检测感染效率,激光共聚焦显微镜观察DC-OVA细胞中Rab4突变体的表达与定位,最后用识别MHC I类分子-OVA肽复合物的T细胞杂交瘤B3Z细胞检测DC-OVA细胞内源抗原递呈的变化。结果慢病毒可高效感染DC-OVA细胞,感染后Rab4突变体可在DC-OVA中稳定表达,主要分布在细胞膜周围的囊泡结构中。抗原递呈检测结果显示Rab4对DC内源性抗原递呈具有正相关性影响。结论成功观察到不同Rab4突变体在DC内的表达与分布,初步证实Rab4蛋白可通过其活性的改变参与调节DC的内源性抗原递呈。
To observe the intracellular localization of Rab4 mutants in dendritic cells and investigate their effects on the endogenous antigen presentation of dendritic cells,we constructed lentiviral vectors of Rab4 mutants and packaged them into 293FT using DNA-Ca3（PO4）2 co-transfection method for producing lentivirus.The lentivirus were used to infected DC-OVA cells and the efficiency of infection was measured by FACS.The expression and location of Rab4 mutants in DC-OVA were observed using confocal laser microscope,and finally the endogenous antigen presentation of DC-OVA was measured with IL-2 level produced by B3Z.The results showed that lentivirus could infect DC-OVA efficiently,and Rab4 mutants could be stably expressed in the organelles around the nucleus.A positive correlation between Rab4 and endogenous antigen presentation of DC was also found.In conclusion,the expression and location of Rab4 mutants have been observed in DC-OVA cells successfully in our study,and we preliminary confirmed that Rab4 could regulate the endogenous antigen presentation of DC by changing its GTPase activity.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2012年第10期877-879,883,共4页
Immunological Journal
关键词
Rab4
DC
突变体
内源
抗原递呈
Rabd
Dendritic cell
Mutant
Endogenous
Antigen presentation
