Preliminary Research On Immunity Response Induced By Intranasal Immune With Inactive Human High Pathogen Avian Influenza Vaccine

Being a new infectious disease, human high pathogen avian influenza(HPAI) has been spread to more than 14 countries. The mortality rate is 63% even higher. It's harm to human health and world economic. WHO required all countries do the prepare for the pandemic influenza. Vaccine is the best way to against infective disease. Current vaccines of HPAI are inactive all virus, splits virus and attenuated virus vaccine. Inactive whole virus has finished I II clinical research. And it has confirmed safety and validity. But the new research find, high pathogen avian influenza virus variant quickly. The vaccines can't stimulate enough immune response to old and children. And the vaccines were performed by injection. It isn't convenient for people. Now it's require a new vaccine has good cross immune protect, easy to performed and suitable to different people.Intranasal immune can induce the systemic and mucosal antibody response better than injection immunity. In this research, we choose inactive pandemic influenza vaccine(Vietnam strain) intranasal immune to observe the Balb/c mouse systemic and mucosal response; and intranasal immune with different pandemic influenza vaccine (Anhui strain, Vietnam strain and Indonesia stain and three stains mixed) to observe the differences in immune responses and provide the theory dependencies of intranasal pandemic influenza vaccine .The study was carried out in three parts:One of the first parts is the preparations of inactive human high pathogen avian influenza antigen. Firstly, choose WHO recommend the H5N1 strain (A/Vietnam/H5N1-PR8/CDC-RG,A/Anhui/1/2005(H5N1)-RG5,A/Indo/05/2005(H5N1)-RG2). and inoculate in eggs. Prepare and identify the antigen of human avian influenza virus strain (H5N1); Secondly, the preparation and purify the antigen. And detect the contents of HA. At last prepare the adjuvant MF59 and mix with the inactive antigen. inactive pandemic influenza vaccine are qualified in WHO demmands.The second part is intranasal immune with inactive pandemic influenza vaccine (A/Vietnam/H5N1-PR8/CDC-RG) . To observe the Balb/c mouse systemic and mucosal response we conform twice immune at 0,14 day and detect the titer of serum IgG IgM, neutral antibody and the cross protect in different strains and subtype viruses, the titer of specific secrete IgG and IgA in lung and nasal washes, observe the dynamic behavior of specific IgA antibody secret cell(ASC) and INF-γ,IL-4 secret cell in spleen lymph cell at 7,14,21,28day after immune. It suggests vaccine can stimulate strong immunity response in serum. HAI antibody is 1: 113±1.5, neutral antibody is 1: 221±82 after 2 weeks with twice immune. The subtype in IgG, IgG1 and IgG2b is the most. The specific secrete IgG and IgA in lung and nasal washes can be detected, sIgA is higher than sIgG. It suggests vaccine can stimulate mucosal immunity. And we also can detect the cell response of Th1, Th2 with boost immunity.The last part, intranasal immune with inactive pandemic influenza vaccine (A/Vietnam/H5N1-PR8/CDC-RG) . To observe the Balb/c mouse systemic and mucosal response we conform twice immune at 0,14 day and detect the titer of serum IgG IgM, neutral antibody and the cross protect in different strains and subtype viruses, the titer of specific secrete IgG and IgA in lung and nasal washes, observe the dynamic behavior of specific IgA antibody secret cell(ASC) and INF-γ, IL-4 secret cell in spleen lymph cell at 7,14,21,28day after immune. It suggests trivalence vaccine can boost stronger immunity than single stain. And the protection is better.Conclusion: the intranasal pandemic influenza vaccine has well immunogenic. It can stimulate systemic and mucosal response and has well crossed immune protection. It provides data to prepare a pandemic influenza vaccine with good cross immune protect, easy to performed and suitable to different people.