Preparation Of The Mycobacterium Tuberculosis Esat-6 Antigen Multi-epitope Dna Vaccine And Their Immune Effects Of A Preliminary Study

PARTⅠConstruction and expression of multiepitope DNA vaccine on ESAT-6 antigen of Mycobacterium tuberculosis in vitroObjective: To construct recombinant plasmids containing 3 T cell epitopes on ESAT-6 antigen of Mycobacterium tuberculosis and extra-cellular fragment of Flt3 ligand (FL) genes, and to express them in rat glomerular mesangial cells (GMCs). Methods: The amino acid sequence of ESAT-6 antigen was analysed using predictive algorithms and 3 T cell epitopes were predicted. The oligonucleotide encoding and the linker Ala-Ala-Tyr (AAY) were synthesized and inserted into the bicistronic vector pIRES and pIRES-FL. The recombinant plasmids were transfected into GMCs, the recombinant proteins expressing in GMCs was examined by Western blot. Results: The recombinant plasmids were verified by sequencing and the recombinant fusion proteins were identified by Western blot. Conclusion: The multiepitope DNA vaccine from ESAT-6 antigen of Mycobacterium tuberculosis was constructed and expressed successfully. PARTⅡImmunol effects of gene vaccine constituted with epitopes of tuberculosis ESAT-6 antigen and Flt-3 ligand in miceObjective: To study the immunol effects of gene vaccine constituted with epitopes of tuberculosis ESAT-6 antigen and Flt-3 ligand in mice. Methods: C57BL/6 mice were inoculated with 100μg of pIRES-TH and pIRES-TH-FL each time and a total of 3 times as a whole. The spleen cell proliferation responses to antigen, the Th1/Th2 responses and the titer of ESAT-6 specific IgG2a/IgGl antibody were examined. Results: The spleen cell proliferation responses, the Th1 responses and the antibody level of ESAT-6 specific IgG2a/IgG1 of the pIRES-TH-FL group are higher than those of control groups. Conclusion: pIRES-TH-FL can effectively elevate the cellular immunity of C57BL/6 mice.