Study On Hsp65for Immunifaction Of Goose Paramyxovirus DNA Vaccine

Goose Newcastle viruse also called goose paramyxovirus, is a serious avian disease. The mortality rate in young geese under15days of age was100%, causing significant economic loses in Goose poultry. Vaccination as one of a effective control measures has been used to keep the disease under control in most countries, traditional vaccines is not have a very ideal protect, novel vaccine research will become central issue. Development of a new vaccine may improve the immunogenicity of NDV DNA vaccine. DNA vaccine may elicit cellular and humoral immune, with advantages of low toxicity. It is necessary to use of adjuvant for improve the immunogenicity of DNA vaccine.The purpose of this research is to explore Mycobacterium avium subsp Paratuberculosis Hsp65to GPMV HN gene vaccine adjuvants role, take MAP hsp65gene and GPMV HN gene cloning to eukaryotic expression vector pVAX1, obtained right recombinant plasmid pVAX1-HN, pVAX1-hsp65-HN, The transfection of this plasmid to Marc-145cells and the recombinant plasmid pVAXl-HN, pVAX1-hsp65-HN can transient expression in Marc-145cell.With respectively and fractionated, take pVAXl-HN, pVAX1-hsp65-HN to goose1000μg per one by intramuscular injection, the antibodytiter by HI as upward trend of immunized geese. The results shows antibodytiter by HI that pVAXl-hsp65-HN are higher than inactivated vaccine and pVAX1-HN at63days. Detection specific antibody level in serum by indirect ELISA, the results shows antibody titer rising with the increasing of days. Inactivated vaccine immunized group to DNA recombinant plasmid specific antibody level in goose serum, difference was significant(P<0.05) before34days. pVAX1-hsp65-HN immunized group and inactivated vaccine GPMV specific antibody level in goose serum, difference was not significant at49days. Both are higher than pVAX1-HN immunized group. Detection T activity by lymphocyte transformation test, the results shows that pVAX1-hsp65-HN T lymphocytes proliferation index are higher pVAXl-HN and inactivated vaccine was not significant(P>0.05), recombinant plasmid pVAX1-HN, pVAX1-hsp65-HN, inactivated vaccine T lymphocytes proliferation index to pVAX1, PBS difference was significant(P<0.05). The above test illustrate that MAP Hsp65enhance immune effect of pVAX1-HN, establish the foundation for goose paramyxovirus DNA vaccine and Hsp65adjuvants role.