| The H9 subtype of avian influenza has endangered our country's poultry industry seriously.In order to prevent and control this avian influenza,our country has researched and developed at least fifteen species of H9 subtype of avian influenza vaccines which come from avian influenza virus.All of these vaccines are inactivated vaccines,and they only could produce antibodies in blood circulation,but could not activate the body to produce mucosal immunity and cellular immunity responses.Due to the variability avian influenza virus,when the vaccines are used in large area,influenza viruses mutate and recombine to escape the pressure which is from the host immune,and emerge the new variants.When the original vaccines face these new variants,they can not provide the effective immune protection,to the new variants,which leads to the new outbreaks of epidemic in clinical or invisible infection and so on.To improve the protective efficacy of avian influenza vaccines and broaden the protective spectrum of vaccines is the key point in the new influenza vaccine research.The regular research and development ways cause the process lengthy.It can not adapt to the changes of epidemiology mutant which emerge constantly.Immune enhancer can improve humoral immunity(including serum antibody and mucosal antibodies),cellular immunity and vaccine effectiveness.From the previous study,we has obtained VA5 immune enhancer can improve the protection about the present inactivated vaccine effectiveness,including humoral and cellular immunity.This study is intended to explore the mechanism of mucosal immunity and cellular immunity of H9 subtypes avian influenza inactivated vaccine which is improved by immunity enhancer.1 The influence of the immunity enhancer to H9 subtype avian influenza inactivated vaccine about IgA and IgG content in mucosa exudates.Method:75 SPF chickens were divided randomly into three groups.25 chickens:Inoculated H9+VA5 mixed vaccine(H9-VA5 group).25 chickens:vaccinated H9 vaccine(H9 group).25 chickens:healthy control group.Each chicken was injected at subcutaneous 0.5ml whereas the the control group C weren't treated.On the third,seventh,fourteenth,and twenty-first day after injection,three chickens were randomly chosen from each group,respectively.After weighing,we collected tears and blood in order to finish the subsequent antibody detection.We collected tracheal mucous,intestinal scraping fluid and lung fluid partly,which were set cryopreservation rapidly and later used as a detection antibody.Results:Immune enhancer VA5 increased the potency of H9 subtype avian influenza inactivated vaccine significantly about 21og2 HI antibody potency.To further evaluate enhancement of the protection effectiveness of H9 avian influenza inactivated vaccine by the immunity enhancer,we used iELISA to test H9+VA5 group,H9 group,and C group on the third,seventh,fourteenth,and twenty-first day.We found that IgG antibody in blood reached a higher level after 21 days,but IgA' level was lower.IgA and IgG antibody's levels were rising with time.On the twenty-first day,IgA content:H9 + VA5 group>H9 group>Group C;IgG content:H9 + VA5 group>H9 group>group C.The IgA's levels in each mucosal exudates on the 21st day:H9 + VA5 group>H9 group>Group C;IgG content:H9 + VA5 group>H9 group>C group.That means the immunity enhancer can enhance the secretion of IgA mucosal antibodies significantly.When we used H9N2 avian influenza virus which was separate source strain to challenge protection,H9+VA5 group's protection was 100%and cloacae swabs and throat swabs never appeared virus.2 The dynamic influence of the immunity enhancer to cells which secrete mucous membrane antibodiesMethod:75 SPF chickens were divided randomly into three groups.25 chickens:Inoculated H9+VA5 mixed vaccine(H9-VA5A group).25 chickens:vaccinated H9 vaccine(H9 group).25 chickens:healthy control group.Each chicken was injected at subcutaneous 0.5mL whereas the control group C weren't treated.On the third,seventh,fourteenth,and twenty-first day after injection,three chickens were randomly chosen from each group,respectively.After weighing we collected lung,spleen,lymph nodes cecum and bursal for testing.Results:In the whole period of immunization,H9+VA5 group,with the progressive of time,IgA and IgG B cells showed an increasing trend,on the twenty-first day the number of B cells got the highest point.And the mostly of IgA and IgG B cells were at the same position.H9 group,with the progressive of time,IgA and IgG B cells also showed an increasing trend,but the number of cells were less than the first group.Control group with the time processed,there was no change on the number of IgA and IgG B cells,less far away from the first and second group.Spleen compared bursa,lung cecal lymph effect more visible.The results show that immunity enhancer VA5 enhances improve H9 subtype inactivated vaccine efficacy through increasing the number of IgA's cells.And the mostly of IgA and IgG B cells are at the same position.3 The influences of the immunity fortifier to H9 subtype of CTL.Method:75 SPF chickens were divided randomly into three groups.25 chickens:Inoculated H9+VA5 mixed vaccine(H9-VA5 A group).25 chickens:vaccinated H9 vaccine(H9 group).25 chickens:healthy control group.Each chicken was injected at subcutaneous 0.5mL whereas the control group C weren't treated.The isolated peripheral blood lymphocytes as effector cells,DF-1 cells infected with the H9 virus as target cells,and the proportion of effector cells and target cells was 25,50,100,200.Results:For H9+VA5 group,with the proportion of effector cells and target cells increasing,the killing rate showed an increasing trend,the proportion of effector cells and target cells reach the highest point when the rate got 200:1.For H9 group,with the proportion of effector cells and target cells increasing,the increased killing rate was not significant.Control group killing rate was substantially zero.H9 + VA5 group and H9 group comparison,the same proportion of effector cells and target cells,H9 + VA5 group killing rate was much higher than H9 group.We can conclude that immunity enhancer through improve the number of IgA cells,thereby increasing the level of mucosal IgA antibodies.Meanwhile,through the improvement of CTL response,it can produce the protection of H9 subtype inactivated vaccine. |
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