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Conference Paper: A pilot study of oral vaccine development against H5N1 Avian Flu using SALMONELLA ASDA mutant as delivery system

TitleA pilot study of oral vaccine development against H5N1 Avian Flu using SALMONELLA ASDA mutant as delivery system
Authors
KeywordsH5N1
Salmonella
AsdA mutant
Issue Date2011
PublisherAkademiai Kiado Rt.. The Journal's web site is located at http://akkrt.hu/73/journals/products/medicine/european_journal_of_microbiology_and_immunology
Citation
The 1st European Conference of Microbiology and Immunology, Budapest, Hungary, 12-14 May 2011. In European Journal of Microbiology and Immunology, 2011, v. 1 n. 2, p. 181-182, abstract no. D10 How to Cite?
AbstractINTRODUCTION: A highly pathogenic avian influenza A virus H5N1 has caused severe disease in human. The major cases have been associated with direct exposure to H5N1 viruses-infected poultry. It has been reported live attenuated Salmonella strains expressed hemaggutinin (HA1) of H5N1 in chicken and showed partial protection versus H5N1 challenge. Salmonella spp. was a Gram-negative and intracellular pathogen. Attenuated Salmonella strains can act as carriers for heterologous antigen delivery due to their efficient mucosal antigen delivery capability. For such purpose, a number of attenuated mutations and antibiotic-free balanced-lethal plasmids were performed in Salmonella spp. Now a considerable concern was arisen for such biological containment systems which included the potential risks for unintentional release of these genetically modified organisms to outer environment. Therefore, development of Salmonella vaccines with high efficacy and self-lysis characteristics for poultry, especially for wild birds, is urgently needed. OBJECTIVE: This project aims to develop a novel live attenuated and self-lysis Salmonella to deliver viral antigens or antiviral shRNAs as oral vaccine for poultry and wild birds. METHOD: Via knocking out asdA gene of a chicken isolated Salmonella, a live attenuated strain (ΔS129) was generated which required 2,6-Diaminoheptanedioic acid (DAP) in growth. After integrating plasmid pEGFP into ΔS129, the expression of eGFP in pEGFP-ΔS129 infected Caco-2 cell was detected. ELISA assay was applied to evaluate the eGFP titer within pEGFP-ΔS129 intraperitoneal administrated Balb/C mice. Flow cytometry detected eGFP expression in peripheral blood mononuclear cell (PBMC) of pEGFP-ΔS129 intragastric administrated Balb/C mice. RESULT: 1. The expression of eGFP in the pEGFP-ΔS129 infected Caco-2 cell can be detected by western blot assay, which displayed a dose-dependent manner for the DAP dosage. 2. ELISA for eGFP detection showed the mean optical densities of 105 CFU group (Dilution Fold=1/50) were 0.3198 (Post Infection Days=6) and 0.3828 (Post Infection Days=23) under 450 nm wave length. 3. Flow cytometry result showed eGFP expression in 0.38% PBMCs, compared with 0.11% in negative control group. CONCLUSION: ΔS129 could be used as a heterologous antigen vehicle in vitro under the regulation of DAP. The in vivo application needs further confirmation in chicken model.
DescriptionPoster abstracts - Session D: Viral Infections and Vaccines
Persistent Identifierhttp://hdl.handle.net/10722/135936
ISSN

 

DC FieldValueLanguage
dc.contributor.authorZhang, Ken_US
dc.contributor.authorLou, YSen_US
dc.contributor.authorZheng, Ben_US
dc.date.accessioned2011-07-27T02:00:09Z-
dc.date.available2011-07-27T02:00:09Z-
dc.date.issued2011en_US
dc.identifier.citationThe 1st European Conference of Microbiology and Immunology, Budapest, Hungary, 12-14 May 2011. In European Journal of Microbiology and Immunology, 2011, v. 1 n. 2, p. 181-182, abstract no. D10en_US
dc.identifier.issn2062-509X-
dc.identifier.urihttp://hdl.handle.net/10722/135936-
dc.descriptionPoster abstracts - Session D: Viral Infections and Vaccines-
dc.description.abstractINTRODUCTION: A highly pathogenic avian influenza A virus H5N1 has caused severe disease in human. The major cases have been associated with direct exposure to H5N1 viruses-infected poultry. It has been reported live attenuated Salmonella strains expressed hemaggutinin (HA1) of H5N1 in chicken and showed partial protection versus H5N1 challenge. Salmonella spp. was a Gram-negative and intracellular pathogen. Attenuated Salmonella strains can act as carriers for heterologous antigen delivery due to their efficient mucosal antigen delivery capability. For such purpose, a number of attenuated mutations and antibiotic-free balanced-lethal plasmids were performed in Salmonella spp. Now a considerable concern was arisen for such biological containment systems which included the potential risks for unintentional release of these genetically modified organisms to outer environment. Therefore, development of Salmonella vaccines with high efficacy and self-lysis characteristics for poultry, especially for wild birds, is urgently needed. OBJECTIVE: This project aims to develop a novel live attenuated and self-lysis Salmonella to deliver viral antigens or antiviral shRNAs as oral vaccine for poultry and wild birds. METHOD: Via knocking out asdA gene of a chicken isolated Salmonella, a live attenuated strain (ΔS129) was generated which required 2,6-Diaminoheptanedioic acid (DAP) in growth. After integrating plasmid pEGFP into ΔS129, the expression of eGFP in pEGFP-ΔS129 infected Caco-2 cell was detected. ELISA assay was applied to evaluate the eGFP titer within pEGFP-ΔS129 intraperitoneal administrated Balb/C mice. Flow cytometry detected eGFP expression in peripheral blood mononuclear cell (PBMC) of pEGFP-ΔS129 intragastric administrated Balb/C mice. RESULT: 1. The expression of eGFP in the pEGFP-ΔS129 infected Caco-2 cell can be detected by western blot assay, which displayed a dose-dependent manner for the DAP dosage. 2. ELISA for eGFP detection showed the mean optical densities of 105 CFU group (Dilution Fold=1/50) were 0.3198 (Post Infection Days=6) and 0.3828 (Post Infection Days=23) under 450 nm wave length. 3. Flow cytometry result showed eGFP expression in 0.38% PBMCs, compared with 0.11% in negative control group. CONCLUSION: ΔS129 could be used as a heterologous antigen vehicle in vitro under the regulation of DAP. The in vivo application needs further confirmation in chicken model.-
dc.languageengen_US
dc.publisherAkademiai Kiado Rt.. The Journal's web site is located at http://akkrt.hu/73/journals/products/medicine/european_journal_of_microbiology_and_immunology-
dc.relation.ispartofEuropean Journal of Microbiology and Immunologyen_US
dc.subjectH5N1-
dc.subjectSalmonella-
dc.subjectAsdA mutant-
dc.titleA pilot study of oral vaccine development against H5N1 Avian Flu using SALMONELLA ASDA mutant as delivery systemen_US
dc.typeConference_Paperen_US
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=2062-509X&volume=1&issue=2&spage=181&epage=182, abstract no. D10&date=2011&atitle=A+pilot+study+of+oral+vaccine+development+against+H5N1+Avian+Flu+using+SALMONELLA+ASDA+mutant+as+delivery+system-
dc.identifier.emailZhang, K: kezhang@hku.hken_US
dc.identifier.emailZheng, B: bzheng@hkucc.hku.hk-
dc.identifier.authorityZheng, B=rp00353en_US
dc.description.naturelink_to_OA_fulltext-
dc.identifier.doi10.1556/EuJMI.1.2011.2.1-
dc.identifier.hkuros188735en_US
dc.identifier.volume1-
dc.identifier.issue2-
dc.identifier.spage181-
dc.identifier.epage182-
dc.description.otherThe 1st European Conference of Microbiology and Immunology, Budapest, Hungary, 12-14 May 2011. In European Journal of Microbiology and Immunology, 2011, v. 1 n. 2, p. 181-182, abstract no. D10-

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