[Construction of Ag85B DNA vaccine for Mycobacterium tuberculosis and its immunogenicities].
OBJECTIVE
To construct pcDNA3-Ag85B recombinant and evaluate its immunogenicity and types of immune response induced by recombinant pcDNA3-Ag85B.


METHODS
Ag85B gene of Mycobacterium tuberculosis was inserted into pcDNA3 to construct DNA vaccine pcDNA3-Ag85B. BALB/c mice were intramuscularly immunized with DNA vaccine, saline and pcDNA3, respectively. The levels of anti-Ag85B antibody were examined with ELISA and the transcripts of IL-2, IL-4, IL-10 and IFN-gamma of spleen lymphocytes stimulated with recombinant Ag85B were tested with RT-PCR at the end of 6th week of inoculation.


RESULTS
Compared with control group, the specific antibody levels against Ag85B were much higher and the IL-2 and IFN-gamma transcripts were up regulated in the pcDNA3-Ag85B group. There was no change of IL-4 and IL-10 expressions in three groups.


CONCLUSIONS
The pcDNA3-Ag85B stimulated the specific antibody products and induced the Th1 type cell immunity.
You
L
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Shandong Cancer Biotherapy Center, Institute of Basic Medicine, Shandong Academy of Medical Sciences, Ji'nan 250062, China.
Zhao
Y
Y
Gao
C
C
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D001428:Bacterial Vaccines / Q000276:immunology*; D019556:COS Cells; D002522:Chlorocebus aethiops; D004269:DNA, Bacterial / Q000276:immunology*; D015870:Gene Expression; D005822:Genetic Vectors; D007111:Immunity, Cellular; D007074:Immunoglobulin G / Q000097:blood; D007371:Interferon-gamma / Q000235:genetics; D016753:Interleukin-10 / Q000235:genetics; D007376:Interleukin-2 / Q000235:genetics; D015847:Interleukin-4 / Q000235:genetics; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000276:immunology; D015183:Restriction Mapping; D019444:Vaccines, DNA / Q000276:immunology*
D000907:Antibodies, Bacterial; D001428:Bacterial Vaccines; D004269:DNA, Bacterial; D007074:Immunoglobulin G; D007376:Interleukin-2; D019444:Vaccines, DNA; D016753:Interleukin-10; D015847:Interleukin-4; D007371:Interferon-gamma