Title: The immune efficacy of OmpA-DNA vaccine against avian pasteurella multocida
Abstract:Objective:To research on protective immunity of OmpA-DNA vaccine against fowl cholera in chickens.Methods:The ompa gene fragment amplified by PCR from avian Pasteurella multocida was cloned into the e...Objective:To research on protective immunity of OmpA-DNA vaccine against fowl cholera in chickens.Methods:The ompa gene fragment amplified by PCR from avian Pasteurella multocida was cloned into the eukaryotic expression vector pcDNA3.1(+),and the recombinant plasmid pOMPA was constructed.Then the recombinant plasmid was transfected into SP2/0 cells in vitro.The transcription and expression of target gene were analyzed by RT-PCR and indirect immunofluorescence.Four groups of chickens named pOMPA,attenuated live vaccine,pCDNA3.1(+) and PBS were vaccinated with the DNA vaccine,attenuated live vaccine,control vector and PBS respectively.The serum antibodies were detected by indirect ELISA.The peripheral blood lymphocyte(PBLC) proliferation and secreted of PBLC were tested by MTT.The immunized chickens were challenged with virulent of avian Pasteurella multocida on week 2 post the third immunization,and the protection rate was counted.Results:RT-PCR and indirect immunofluorescence showed that the ompa gene could be transfected into SP2/0 cells in vitro and the target protein was expressed.Indirect ELISA showed that the levels of antibodies in DNA vaccine group and attenuated live vaccine group were most significantly higher than those of the other groups(P0.01),and that of the former lower than that of the latter(P0.05).MTT experiments indicated that the SI value induced with avian Pasteurella multocida Omps in the two vaccine groups were significantly higher than those of pCDNA3.1(+) and PBS groups(P0.01),and the two vaccines groups were indiscriminate.The IFN-γ experiments displayed that the levels of IFN-γ induced with Omps in the groups of pOMPA and attenuated vaccine were mostly higher than those of the control groups(P0.01).The protection rate of DNA vaccine and attenuated live vaccine were 60% and 73.3% respectively.Conclusion:The OmpA-DNA vaccine against fowl cholera has been constructed successfully.The DNA vaccine could enhance the immunity level and the protective effect against fowl cholera,but is barely satisfactory.Read More
Publication Year: 2011
Publication Date: 2011-01-01
Language: en
Type: article
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