ABSTRACT
The aim of this study is to investigate the immune effects of BCG primary immunization and IL-12 combined with Mycobacterium tuberculosis Ag85A DNA vaccine booster immunization on mice .We randomly divided the mice into 7 groups ,namely PBS negative controls ,BCG controls ,pcAg85A controls ,BCG primary immunization combined with Ag85A booster immunization controls ,BCG primary immunization combined with Ag85A and IL-12 booster immunization controls , BCG primary immunization combined with IL-12 booster immunization controls ,and BCG primary immunization combined with pcDNA3 .1 booster immunization controls .Implementing the immune in procedure of BCG primary immunization and cytokine IL-12 booster immunization combined with mycobacterium tuberculosis Ag85A DNA ,we observed the immune effect on mice by detecting the mice serum total IgG , specific lymphocyte proliferation and cytokine levels in 4 ,6 ,8 weeks after the last immunization .Comparing the mice immunized in the strategy of BCG primary immunization and cytokine IL-12 combined with Mycobacterium tuberculosis Ag85A DNA vaccine strengthening immunization to the mice in other groups by other immune ways ,we found that ,in BCG/Ag85A+ IL-12 groups ,IgG in-creased significantly (P＜ 0 .05) ,specific lymphocyte prolifera-ted significantly ,and after strengthening immunization IFN-γlevels ,IL-2 levels and IL-4 levels in the three periods were 128 .2 ±20.4,190.2±16.51,244.2±39.14 ;146.2±17.29,271.6±16.36and16.36±28.12 ;68.6±6.62,96.6±5.5and5.5± 10 .71 ,respectively ,which were higher than those in other groups (P＜0 .05) .It’s suggested that the immunization way of BCG primary immunization and cytokine IL-12 combined with Mycobacterium tuberculosis Ag85A DNA vaccine booster immu-nization could significantly enhance the humoral and cellular immunity of the bodies ,and provide the basis for further study on protective effect test in animals .
ABSTRACT
Objective:To investigate the immune efficacy after sequential immunization with Mycobacterium tuberculosis DNA vaccine encoding mature form of Ag85B(pTB30m)and Mycobacterium tuberculosis H37Ra in mice.Methods:Much of highly pure plasmid DNA(pTB30m)extracted by alkaline lysis method was confirmed by restriction endonuclease digestion.Then,its DNA concentration and purity were determined by UV spectrophotometry.At various intervals(4weeks,8weeks)after sequential immunization,ELISA was used to detect the level of the serum antibody against PPD.Also,the spleen lymphocytes of mice were cultured with PPD in vitro.Lymphocyte transformation was detected by MTT assay.Results:Prepared pTB30m was highly pure and came to the needed concentration.Compared with Group Naive control,the specific antibody levels against PPD and the stimulation index(SI)of spleen lymphocytes were all statistically higher in Group DNA-85B/H37Ra(P0.05),as compared with Group DNA-85B/BCG,Group H37Ra and Group BCG,However,compared with Group H37Ra and Group BCG,the SI of mice was significantly larger in Group DNA-85B/H37Ra(P