Autophagy-targeted DNA vaccine against Japanese encephalitis virus promotes dendritic cell function in BALB/c mice
10.3760/cma.j.cn112309-20201024-00488
- VernacularTitle:增强自噬的乙型脑炎重组DNA疫苗促进BALB/c小鼠树突状细胞功能
- Author:
Junyao ZHU
;
Xin ZANG
;
Yongzhen ZHAI
- From:
Chinese Journal of Microbiology and Immunology
2021;41(3):209-215
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effects of autophagy-targeted DNA vaccine against Japanese encephalitis virus (JEV) on the immune-related functions of dendritic cells (DCs) in BALB/c mice.Methods:Healthy female BALB/c mice were randomly divided into 5 groups and injected with pcDNA3.1(+ ) empty vector (negative control), pJME plasmid, recombinant pJME-LC3 plasmid muscle, sterile PBS (blank control group) and live attenuated JEV vaccine (positive control group), respectively. The mice were immunized three times with an interval of two weeks. Splenic DCs were isolated two weeks after the last immunization. Immunofluorescence assay was used to observe the expression of the recombinant plasmid in dendritic cells. Expression of major histocompatibility complex Ⅱ (MHC Ⅱ) on DCs and the uptake of FITC-Dextran by DCs were observed by flow cytometry. CCK8 assay was used to detect the effects of DCs on the proliferation of spleen mononuclear cells from allogeneic mice.Results:The expression of plasmid-encoded protein in the DCs of the pJME-LC3 group was significantly higher than that of the pJME, pJME-LC3+ 3-MA and pcDNA3.1(+ ) empty vector groups. The uptake of FITC-Dextran by DCs was significantly enhanced in the pJME-LC3 group than in the other groups ( P<0.05), and the expression of MHC Ⅱ moleculars on DCs was increased in the pJME-LC3 group as well ( P<0.05). The splenic DCs from the mice in the pJME-LC3 group had a stronger effect on the proliferation of spleen mononuclear cells from allogeneic mice that those from other groups ( P<0.05). Conclusions:The recombinant plasmid pJME-LC3 could promote the ability of mouse splenic DCs to present antigen and to stimulate lymphocyte proliferation after immunization, suggesting that the autophagy-targeted recombinant DNA vaccine against JEV could enhance immune responses by affecting the function of DCs in BALB/c mice.
