IFN-gamma increases efficiency of DNA vaccine in protecting ducks against infection.
OBJECTIVE
To detect the effects of DNA vaccines in combination with duck IFN-gamma gene on the protection of ducks against duck hepatitis B virus (DHBV) infection.


METHODS
DuIFN-gamma cDNA was cloned and expressed in COS-7 cells, and the antiviral activity of DuIFN-gamma was detected and neutralized by specific antibodies. Ducks were vaccinated with DHBpreS/S DNA alone or co-immunized with plasmid expressing DuIFN-gamma. DuIFN-gamma mRNA in peripheral blood mononuclear cells (PBMCs) from immunized ducks was detected by semi-quantitative competitive RT-PCR. Anti-DHBpreS was titrated by enzyme-linked immunosorbent assay (ELISA). DHBV DNA in sera and liver was detected by Southern blot hybridization, after ducks were challenged with high doses of DHBV.


RESULTS
DuIFN-gamma expressed by COS-7 was able to protect duck fibroblasts against vesicular stomatitis virus (VSV) infection in a dose-dependent fashion, and anti-DuIFN-gamma antibodies neutralized the antiviral effects. DuIFN-gamma in the supernatant also inhibited the release of DHBV DNA from LMH-D2 cells. When ducks were co-immunized with DNA vaccine expressing DHBpreS/S and DuIFN-gamma gene as an adjuvant, the level of DuIFN-gamma mRNA in PBMCs was higher than that in ducks vaccinated with DHBpreS/S DNA alone. However, the titer of anti-DHBpreS elicited by DHBpreS/S DNA alone was higher than that co-immunized with DuIFN-gamma gene and DHBpreS/S DNA. After being challenged with DHBV at high doses, the load of DHBV in sera dropped faster, and the amount of total DNA and cccDNA in the liver decreased more significantly in the group of ducks co-immunized with DuIFN-gamma gene and DHBpreS/S DNA than in other groups.


CONCLUSIONS
DHBV preS/S DNA vaccine can protect ducks against DHBV infection, DuIFN-gamma gene as an immune adjuvant enhances its efficacy.
Long
Jian-Er
JE
Department of Molecular Virology, Shanghai Medical College, Fudan University, Shanghai 200032, China.
Huang
Li-Na
LN
Qin
Zhi-Qiang
ZQ
Wang
Wen-Yi
WY
Qu
Di
D
D000276:Adjuvants, Immunologic / Q000235:genetics; D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D004195:Disease Models, Animal; D004372:Ducks; D018347:Hepadnaviridae Infections / Q000276:immunology / Q000517:prevention & control*; D015611:Hepatitis B Virus, Duck; D006524:Hepatitis, Viral, Animal / Q000276:immunology / Q000517:prevention & control*; D007371:Interferon-gamma / Q000235:genetics*; D019444:Vaccines, DNA / Q000235:genetics*
D000276:Adjuvants, Immunologic; D019444:Vaccines, DNA; D007371:Interferon-gamma