Limits...
Construction and Immunogenicity Evaluation of Recombinant Influenza A Viruses Containing Chimeric Hemagglutinin Genes Derived from Genetically Divergent Influenza A H1N1 Subtype Viruses.

McCormick K, Jiang Z, Zhu L, Lawson SR, Langenhorst R, Ransburgh R, Brunick C, Tracy MC, Hurtig HR, Mabee LM, Mingo M, Li Y, Webby RJ, Huber VC, Fang Y - PLoS ONE (2015)

Bottom Line: Nine shuffled HA constructs were initially screened for immunogenicity in mice by DNA immunization, and one chimeric HA (HA-129) was expressed on both a A/Puerto Rico/8/34 backbone with mutations associated with a live, attenuated phenotype (PR8LAIV-129) and a A/swine/Texas/4199-2/98 backbone (TX98-129).When delivered to mice, the PR8LAIV-129 induced antibodies against all four parental viruses, which was similar to the breadth of immunity observed when HA-129 was delivered as a DNA vaccine.This study established a platform for creating novel genes of influenza viruses using a molecular breeding approach, which will have important applications toward future development of broadly protective influenza virus vaccines.

View Article: PubMed Central - PubMed

Affiliation: Division of Basic Biomedical Sciences, Sanford School of Medicine, The University of South Dakota, Vermillion, SD, 57069, United States of America.

ABSTRACT

Background and objectives: Influenza A viruses cause highly contagious diseases in a variety of hosts, including humans and pigs. To develop a vaccine that can be broadly effective against genetically divergent strains of the virus, in this study we employed molecular breeding (DNA shuffling) technology to create a panel of chimeric HA genes.

Methods and results: Each chimeric HA gene contained genetic elements from parental swine influenza A viruses that had a history of zoonotic transmission, and also from a 2009 pandemic virus. Each parental virus represents a major phylogenetic clade of influenza A H1N1 viruses. Nine shuffled HA constructs were initially screened for immunogenicity in mice by DNA immunization, and one chimeric HA (HA-129) was expressed on both a A/Puerto Rico/8/34 backbone with mutations associated with a live, attenuated phenotype (PR8LAIV-129) and a A/swine/Texas/4199-2/98 backbone (TX98-129). When delivered to mice, the PR8LAIV-129 induced antibodies against all four parental viruses, which was similar to the breadth of immunity observed when HA-129 was delivered as a DNA vaccine. This chimeric HA was then tested as a candidate vaccine in a nursery pig model, using inactivated TX98-129 virus as the backbone. The results demonstrate that pigs immunized with HA-129 developed antibodies against all four parental viruses, as well as additional primary swine H1N1 influenza virus field isolates.

Conclusion: This study established a platform for creating novel genes of influenza viruses using a molecular breeding approach, which will have important applications toward future development of broadly protective influenza virus vaccines.

No MeSH data available.


Related in: MedlinePlus

IgG antibody response in mice immunized with plasmid DNAs expressing chimeric HA.Mice (n = 4) were vaccinated with plasmid DNAs of chimeric HA, delivered by gene gun. Serum antibody (IgG) titers after third vaccination were evaluated by ELISA, with samples considered positive if their initial serum OD405 was at least three times greater than the OD405 of negative control sera. Samples with antibody titers below the detectable limit of the assay were assigned a titer of 50 for the purpose of generating graphs. Horizontal bars show mean values, and vertical error bars indicate standard deviation.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4465703&req=5

pone.0127649.g003: IgG antibody response in mice immunized with plasmid DNAs expressing chimeric HA.Mice (n = 4) were vaccinated with plasmid DNAs of chimeric HA, delivered by gene gun. Serum antibody (IgG) titers after third vaccination were evaluated by ELISA, with samples considered positive if their initial serum OD405 was at least three times greater than the OD405 of negative control sera. Samples with antibody titers below the detectable limit of the assay were assigned a titer of 50 for the purpose of generating graphs. Horizontal bars show mean values, and vertical error bars indicate standard deviation.

Mentions: After screening the HA composition, selected chimeric HAs were screened in mice using DNA immunization. Serum samples collected at 14 days after a third inoculation with DNA were tested for antibody responses using an ELISA that incorporated parental HA-expressing viruses as antigen. The results show that IgG antibodies against all four parental viruses were detected in constructs HA-107, HA-111, HA-113, HA-116, HA-123, and HA-129 (Fig 3). Of note, the HA-124, HA-126, and HA-208 chimeras did not induce antibodies that consistently reacted with all four parental viruses. These data demonstrate that chimeric HA constructs created using DNA shuffling method have the ability to induce broad antibody responses, with some of these constructs inducing antibodies that react with all four parental HAs.


Construction and Immunogenicity Evaluation of Recombinant Influenza A Viruses Containing Chimeric Hemagglutinin Genes Derived from Genetically Divergent Influenza A H1N1 Subtype Viruses.

McCormick K, Jiang Z, Zhu L, Lawson SR, Langenhorst R, Ransburgh R, Brunick C, Tracy MC, Hurtig HR, Mabee LM, Mingo M, Li Y, Webby RJ, Huber VC, Fang Y - PLoS ONE (2015)

IgG antibody response in mice immunized with plasmid DNAs expressing chimeric HA.Mice (n = 4) were vaccinated with plasmid DNAs of chimeric HA, delivered by gene gun. Serum antibody (IgG) titers after third vaccination were evaluated by ELISA, with samples considered positive if their initial serum OD405 was at least three times greater than the OD405 of negative control sera. Samples with antibody titers below the detectable limit of the assay were assigned a titer of 50 for the purpose of generating graphs. Horizontal bars show mean values, and vertical error bars indicate standard deviation.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4465703&req=5

pone.0127649.g003: IgG antibody response in mice immunized with plasmid DNAs expressing chimeric HA.Mice (n = 4) were vaccinated with plasmid DNAs of chimeric HA, delivered by gene gun. Serum antibody (IgG) titers after third vaccination were evaluated by ELISA, with samples considered positive if their initial serum OD405 was at least three times greater than the OD405 of negative control sera. Samples with antibody titers below the detectable limit of the assay were assigned a titer of 50 for the purpose of generating graphs. Horizontal bars show mean values, and vertical error bars indicate standard deviation.
Mentions: After screening the HA composition, selected chimeric HAs were screened in mice using DNA immunization. Serum samples collected at 14 days after a third inoculation with DNA were tested for antibody responses using an ELISA that incorporated parental HA-expressing viruses as antigen. The results show that IgG antibodies against all four parental viruses were detected in constructs HA-107, HA-111, HA-113, HA-116, HA-123, and HA-129 (Fig 3). Of note, the HA-124, HA-126, and HA-208 chimeras did not induce antibodies that consistently reacted with all four parental viruses. These data demonstrate that chimeric HA constructs created using DNA shuffling method have the ability to induce broad antibody responses, with some of these constructs inducing antibodies that react with all four parental HAs.

Bottom Line: Nine shuffled HA constructs were initially screened for immunogenicity in mice by DNA immunization, and one chimeric HA (HA-129) was expressed on both a A/Puerto Rico/8/34 backbone with mutations associated with a live, attenuated phenotype (PR8LAIV-129) and a A/swine/Texas/4199-2/98 backbone (TX98-129).When delivered to mice, the PR8LAIV-129 induced antibodies against all four parental viruses, which was similar to the breadth of immunity observed when HA-129 was delivered as a DNA vaccine.This study established a platform for creating novel genes of influenza viruses using a molecular breeding approach, which will have important applications toward future development of broadly protective influenza virus vaccines.

View Article: PubMed Central - PubMed

Affiliation: Division of Basic Biomedical Sciences, Sanford School of Medicine, The University of South Dakota, Vermillion, SD, 57069, United States of America.

ABSTRACT

Background and objectives: Influenza A viruses cause highly contagious diseases in a variety of hosts, including humans and pigs. To develop a vaccine that can be broadly effective against genetically divergent strains of the virus, in this study we employed molecular breeding (DNA shuffling) technology to create a panel of chimeric HA genes.

Methods and results: Each chimeric HA gene contained genetic elements from parental swine influenza A viruses that had a history of zoonotic transmission, and also from a 2009 pandemic virus. Each parental virus represents a major phylogenetic clade of influenza A H1N1 viruses. Nine shuffled HA constructs were initially screened for immunogenicity in mice by DNA immunization, and one chimeric HA (HA-129) was expressed on both a A/Puerto Rico/8/34 backbone with mutations associated with a live, attenuated phenotype (PR8LAIV-129) and a A/swine/Texas/4199-2/98 backbone (TX98-129). When delivered to mice, the PR8LAIV-129 induced antibodies against all four parental viruses, which was similar to the breadth of immunity observed when HA-129 was delivered as a DNA vaccine. This chimeric HA was then tested as a candidate vaccine in a nursery pig model, using inactivated TX98-129 virus as the backbone. The results demonstrate that pigs immunized with HA-129 developed antibodies against all four parental viruses, as well as additional primary swine H1N1 influenza virus field isolates.

Conclusion: This study established a platform for creating novel genes of influenza viruses using a molecular breeding approach, which will have important applications toward future development of broadly protective influenza virus vaccines.

No MeSH data available.


Related in: MedlinePlus