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Fowl Adenovirus Subunit Vaccine With Molecular Adjuvants

Posted on:2020-08-04Degree:MasterType:Thesis
Country:ChinaCandidate:G R JiFull Text:PDF
GTID:2393330572485758Subject:Biological engineering
Abstract/Summary:PDF Full Text Request
Hydropericardium-hepatitis syndrome and inclusion body hepatitis induced by fowl adenoviruses have caused huge economic losses to the breeding industry in China.Infection of fowl adenoviruses has no obvious clinical symptoms in the early stage so it can not be treated effectively.Therefore,vaccination has become the preferred strategy.This work aims to develop an effective and safe subunit vaccine.According to the etiology of fowl adenovirus,the protein Fiber-2 was chosen to be the antigenic protein.In order to improve the immune efficiency,IL-2 and IFN-? were selected as vaccine adjuvants.The efficacy of the vaccine was evaluated after co-immunization.Firstly,Fiber-2 was produced in the baculovirus expression system.The codon-optimized Fiber-2 sequence was inserted into the baculovirus vector Bacmid.The recombinant plasmid was transfected into Sf9 cells to produce the high titer virus and the recombinant protein.The recombinant protein showed great immunogenicity with the titer up to 1:128.Secondly,IL-2 and IFN-? were expressed in the E.coli system.The codon-optimized sequence was inserted into the expression vector pET28a(+).The recombinant protein was found in the pellet of cell lysate with the form of inclusion bodies.The renaturation results showed that the recombinant chIL-2 and chIFN-? proteins were soluble in the buffer.Finally,different doses of vaccines were injected chickens for animal experiments.The immunoprotection results showed that Fiber-2 could provide 100%protection with a dose of 50?g.The co-immunization results showed that chIFN-? is more effective than chIL-2,and the protection rate of the two adiuvants vaccine reaches 100%,which contains both of adjuvants and the low dose of Fiber-2 protein(10 ?g per dose).
Keywords/Search Tags:Fowl adenoviruses, Fiber-2, IL-2, IFN-?, subunit vaccine, molecular adjuvant
PDF Full Text Request
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