Sequential Immunization With Mycobacterial Antigen Ag85B-Expressing DNA And Mycobacterium Tuberculosis H37Ra In Mice

Objectives: 1. To prepare highly pure DNA vaccine encoding mature form of Ag85B protein of Mycobacterium tuberculosis, namely pTB30m, which was as the priming vaccine in DNA-85B/H37Ra prime-boost vaccination regimen. 2. To investigate the immune efficacy after sequential immunization with mycobacterial antigen Ag85B-expressing DNA(pTB30m)and Mycobacterium tuberculosis H37Ra.Methods: 1. Much of highly pure plasmid DNA(pTB30m)was extracted by alkaline lysis method and was confirmed by restriction endonuclease digestion.Then, its DNA concentration and purity were determined by UV spectrophotometry. 2. C57BL/6 female mice were primed with an intramuscular injecton of a DNA vaccine(pTB30m)and boosted with H37Ra at 2-week intervals in Group DNA-85B/H37Ra. Meanwhile, the others of C57BL/6 female mice were divided into Group DNA-85B/BCG, Group H37Ra, Group BCG and Group Naive control respectively. 3. At various intervals(4 weeks, 8 weeks)after sequential immunization, ELISA was used to detect the level of the serum antibody...