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Bruce H Janke - One of the best experts on this subject based on the ideXlab platform.
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dna vaccination elicits protective immune responses against pandemic and classic Swine Influenza viruses in pigs
Clinical and Vaccine Immunology, 2011Co-Authors: Patrick J Gorres, Kelly M Lager, Wing Pui Kong, Michael Royals, John Paul Todd, Amy L Vincent, Chih Jen Wei, Crystal L Loving, Eraldo Lourenso Zanella, Bruce H JankeAbstract:Swine Influenza is a highly contagious viral infection in pigs that significantly impacts the pork industry due to weight loss and secondary infections. There is also the potential of a significant threat to public health, as was seen in 2009 when the pandemic H1N1 Influenza virus strain emerged from reassortment events among avian, Swine, and human Influenza viruses within pigs. As classic and pandemic H1N1 strains now circulate in Swine, an effective vaccine may be the best strategy to protect the pork industry and public health. Current inactivated-virus vaccines available for Swine Influenza protect only against viral strains closely related to the vaccine strain, and egg-based production of these vaccines is insufficient to respond to large outbreaks. DNA vaccines are a promising alternative since they can potentially induce broad-based protection with more efficient production methods. In this study we evaluated the potentials of monovalent and trivalent DNA vaccine constructs to (i) elicit both humoral and gamma interferon (IFN-γ) responses and (ii) protect pigs against viral shedding and lung disease after challenge with pandemic H1N1 or classic Swine H1N1 Influenza virus. We also compared the efficiency of a needle-free vaccine delivery method to that of a conventional needle/syringe injection. We report that DNA vaccination elicits robust serum antibody and cellular responses after three immunizations and confers significant protection against Influenza virus challenge. Needle-free delivery elicited improved antibody responses with the same efficiency as conventional injection and should be considered for development as a practical alternative for vaccine administration.
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inhibition test in detecting serum antibody against Swine Influenza viruses comparison of a commercial h1n1 enzyme linked immunosorbent assay and hemagglutination
2011Co-Authors: Kyoungjin Yoon, Bruce H Janke, Rick W Swalla, Gene EricksonAbstract:Recently a commercial enzyme-linked immunosorbent assay (ELISA) kit for detecting antibody against H1N1 Swine Influenza virus (SIV) has been made available to diagnosticians and veterinary practitioners. Because the hemagglutination inhibition (HI) test has been considered the standard test for SIV serology, diagnostic performance of the new ELISA was evaluated using positive (n 5 60) and negative (n 5 188) serum samples from young pigs with known status of SIV infection and compared with that of the HI test. Both ELISA and HI test identified all negative animals correctly. None of the serum samples (n 5 64) from pigs inoculated with H3N2 SIV was positive by ELISA for SIV antibody. The H1N1 SIV antibody detectable by ELISA appears to develop more slowly in comparison with antibody detectable by HI test. Although antibody was detected by HI test in all inoculated animals (n 5 20) by day 7 postinoculation (PI), antibody was detected by ELISA in 0%, 75%, and 100% of the inoculated animals on days 7, 14, and 28 PI, respectively. Discrepancy in test results between the 2 serologic tests appeared to be because of differences in antibody isotypes detected by each test. Enzyme-linked immunosorbent assay mainly detected IgG antibody, whereas the HI test detects IgM antibody very efficiently as well as IgG antibody. Collectively, the commercial ELISA is highly specific for antibody to H1N1 SIV but may not identify positive animals at the early stage of infection as effectively as the HI test, particularly when SIV is introduced to a nai ve Swine population. Swine Influenza, commonly known as ‘‘Swine flu,’’ has become one of the economically significant respiratory diseases in pigs throughout the world because it was initially recognized in the early 1900s.5,6,11,16 The disease is caused mainly by Influenza A viruses, which are enveloped RNA viruses with 8-segmented, singlestranded, negative-sense RNA molecules.8 Although detection of Swine Influenza virus (SIV) or viral antigen in the lung or nasal secretions from clinically affected animals is considered as the definitive diagnosis of Swine Influenza, serologic testing is often used to detect animals that have been exposed to SIV because the disease has a very short course and the causative agent becomes undetectable in nasal secretions or lungs relatively quickly.9,17 Serology is also used to assess immune status of pigs at various stages within an operation so that the level of herd immunity or timing of vaccination can be determined. Several serologic assays have been used for detecting antibody against SIV: hemagglutination inhibition (HI) test, serum–virus neutralization test, and indirect From the Veterinary Diagnostic Laboratory, Iowa State University, Ames, IA 50011 (Yoon, Janke), Murphy Family Farms, Algona, IA 50511 (Swalla), and Rollins Animal Disease Diagnostic Laboratory, Raleigh, NC 27605 (Erickson). 1Corresponding Author: Kyoung-Jin Yoon, Associate Professor and Head of Virology, Veterinary Diagnostic Laboratory, College of Veterinary Medicine, Iowa State University, 1600 South 16th Street, Ames, IA 50011. fluorescent antibody test.8,17 Among these assays, HI test has been used most commonly in veterinary diagnostic laboratories to detect anti-Influenza virus antibody and is considered to be the standard test for international trade of animals by Office International des Epizooties.12 The HI test is designed to detect antibody specific for hemagglutinin (H), which is 1 of 2 major envelope proteins on the surface of SIV and highly immunogenic. Because 15 different H subtypes are known to exist among Influenza A viruses,8 the HI test must be tailored for each subtype by using reference strains corresponding to individual subtypes in assays for measuring subtype-specific antibody. At present, H1 and H3 subtypes are of worldwide concern in the Swine industry,1,2,13 although pigs are known to be susceptible to all H subtypes.12 The HI test is a relatively inexpensive serologic assay. In general, the presence of HI antibody in animals is indicative of protection against the subtype used in the test, and HI antibody titers appear to correlate with the level of protection. However, the labor intensiveness of the HI test is a major hindrance to its use on a large scale. Recently, a commercial enzyme-linked immunosorbent assay (ELISA) has been developed specifically for detecting antibody against SIV of H1N1 subtype. The following study was conducted to assess the diagnostic performance of the ELISA in comparison with the HI test using a set of serum samples from animals with known status of Swine Influenza. by guest on May 22, 2011 vdi.sagepub.com Downloaded from
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viral reassortment and transmission after co infection of pigs with classical h1n1 and triple reassortant h3n2 Swine Influenza viruses
Journal of General Virology, 2010Co-Authors: Kelly M Lager, Bruce H Janke, Adolfo Garciasastre, Jurgen A Richt, Porntippa Lekcharoensuk, Alicia Solorzano, Richard J Webby, Eva S UleryAbstract:Triple-reassortant Swine Influenza viruses circulating in North American pigs contain the internal genes derived from Swine (matrix, non-structural and nucleoprotein), human [polymerase basic 1 (PB1)] and avian (polymerase acidic and PB2) Influenza viruses forming a constellation of genes that is well conserved and is called the triple-reassortant internal gene (TRIG) cassette. In contrast, the external genes [haemagglutinin (HA) and neuraminidase (NA)] are less conserved, reflecting multiple reassortant events that have produced viruses with different combinations of HA and NA genes. This study hypothesized that maintenance of the TRIG cassette confers a selective advantage to the virus. To test this hypothesis, pigs were co-infected with the triple-reassortant H3N2 A/Swine/Texas/4199-2/98 (Tx/98) and the classical H1N1 A/Swine/Iowa/15/1930 viruses and co-housed with a group of sentinel animals. This direct contact group was subsequently moved into contact with a second group of naive animals. Four different subtypes (H1N1, H1N2, H3N1 and H3N2) of Influenza virus were identified in bronchoalveolar lavage fluid collected from the lungs of the experimentally infected pigs, with most of the viruses containing TRIG from the Tx/98 virus. Interestingly, only the intact H3N2 Tx/98 virus was transmitted from the infected pigs to the direct-contact animals and from them to the second contact group of pigs. These results demonstrated that multiple reassortments can occur within a host; however, only specific gene constellations are readily transmissible. It was concluded that certain HA and NA gene pairs, in conjunction with the TRIG cassette, may have a competitive advantage over other combinations for transmission and maintenance in Swine.
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efficacy of inactivated Swine Influenza virus vaccines against the 2009 a h1n1 Influenza virus in pigs
Vaccine, 2010Co-Authors: Amy L Vincent, Bruce H Janke, Eraldo L Zanella, Janice R Ciaccizanella, Marcus E Kehrli, Alessio Lorusso, P C Gauger, Kelly M LagerAbstract:The gene constellation of the 2009 pandemic A/H1N1 virus is a unique combination from Swine Influenza A viruses (SIV) of North American and Eurasian lineages, but prior to April 2009 had never before been identified in Swine or other species. Although its hemagglutinin gene is related to North American H1 SIV, it is unknown if vaccines currently used in U.S. Swine would cross-protect against infection with the pandemic A/H1N1. The objective of this study was to evaluate the efficacy of inactivated vaccines prepared with North American Swine Influenza viruses as well as an experimental homologous A/H1N1 vaccine to prevent infection and disease from 2009 pandemic A/H1N1. All vaccines tested provided partial protection ranging from reduction of pneumonia lesions to significant reduction in virus replication in the lung and nose. The multivalent vaccines demonstrated partial protection; however, none was able to prevent all nasal shedding or clinical disease. An experimental homologous 2009 A/H1N1 monovalent vaccine provided optimal protection with no virus detected from nose or lung at any time point in addition to amelioration of clinical disease. Based on cross-protection demonstrated with the vaccines evaluated in this study, the U.S. Swine herd likely has significant immunity to the 2009 A/H1N1 from prior vaccination or natural exposure. However, consideration should be given for development of monovalent homologous vaccines to best protect the Swine population thus limiting shedding and the potential transmission of 2009 A/H1N1 from pigs to people.
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experimental inoculation of pigs with pandemic h1n1 2009 virus and hi cross reactivity with contemporary Swine Influenza virus antisera
Influenza and Other Respiratory Viruses, 2010Co-Authors: Amy L Vincent, Kelly M Lager, Kay S Faaberg, Michelle Harland, Eraldo L Zanella, Janice R Ciaccizanella, Marcus E Kehrli, Bruce H JankeAbstract:Please cite this paper as: Vincent et al. (2010) Experimental inoculation of pigs with pandemic H1N1 2009 virus and HI cross-reactivity with contemporary Swine Influenza virus antisera. Influenza and Other Respiratory Viruses 4(2), 53–60 Background A novel A/H1N1 was identified in the human population in North America in April 2009. The gene constellation of the virus was a combination from Swine Influenza A viruses (SIV) of North American and Eurasian lineages that had never before been identified in Swine or other species. Objectives The objectives were to (i) evaluate the clinical response of Swine following experimental inoculation with pandemic H1N1 2009; (ii) assess serologic cross-reactivity between H1N1 2009 and contemporary SIV antisera; and (iii) develop a molecular assay to differentiate North American-lineage SIV from H1N1 2009. Methods Experiment 1: Weaned pigs were experimentally infected with A/California/04/2009 (H1N1). Experiment 2: The cross-reactivity of a panel of US SIV H1N1 or H1N2 antisera with three isolates of pandemic A/H1N1 was evaluated. Experiment 3: A polymerase chain reaction (PCR)-based diagnostic test was developed and validated on samples from experimentally infected pigs. Results and Conclusions In experiment 1, all inoculated pigs demonstrated clinical signs and lesions similar to those induced by endemic SIV. Viable virus and antigen were only detected in the respiratory tract. In experiment 2, serologic cross-reactivity was limited against H1N1 2009 isolates, notably among virus antisera from the same HA phylogenetic cluster. The limited cross-reactivity suggests North American pigs may not be fully protected against H1N1 2009 from previous exposure or vaccination and novel tests are needed to rapidly diagnose the introduction of H1N1 2009. In experiment 3, an RT–PCR test that discriminates between H1N1 2009 and endemic North American SIV was developed and validated on clinical samples.
Amy L Vincent - One of the best experts on this subject based on the ideXlab platform.
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divergent immune responses and disease outcomes in piglets immunized with inactivated and attenuated h3n2 Swine Influenza vaccines in the presence of maternally derived antibodies
Virology, 2014Co-Authors: Matthew R Sandbulte, Crystal L Loving, Ratree Platt, James A Roth, Jamie Henningson, Kathleen A Gibson, Daniela S Rajao, Amy L VincentAbstract:Live-attenuated Influenza virus (LAIV) prime-boost vaccination previously conferred protection against heterologous H3N2 Swine Influenza challenge, including in piglets with maternally derived antibodies (MDA). Conversely, a whole-inactivated virus (WIV) vaccine was associated with enhanced disease. This study was aimed at identifying immune correlates of cross-protection. Piglets with and without MDA received intramuscular adjuvanted WIV or intranasal LAIV, and were challenged with heterologous H3N2. WIV induced cross-reactive IgG, inhibited by MDA, and a moderate T cell response. LAIV elicited mucosal antibodies and T cells cross-reactive to the heterologous challenge strain. The presence of MDA at LAIV vaccination blocked lung and nasal antibody production, but did not interfere with T cell priming. Even without mucosal antibodies, MDA-positive LAIV vaccinates were protected, indicating a likely role for T cells. Based on the data, one LAIV dose can induce cell-mediated immunity against antigenically divergent H3N2 Influenza virus despite passive antibody interference with humoral immune responses.
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dna vaccination elicits protective immune responses against pandemic and classic Swine Influenza viruses in pigs
Clinical and Vaccine Immunology, 2011Co-Authors: Patrick J Gorres, Kelly M Lager, Wing Pui Kong, Michael Royals, John Paul Todd, Amy L Vincent, Chih Jen Wei, Crystal L Loving, Eraldo Lourenso Zanella, Bruce H JankeAbstract:Swine Influenza is a highly contagious viral infection in pigs that significantly impacts the pork industry due to weight loss and secondary infections. There is also the potential of a significant threat to public health, as was seen in 2009 when the pandemic H1N1 Influenza virus strain emerged from reassortment events among avian, Swine, and human Influenza viruses within pigs. As classic and pandemic H1N1 strains now circulate in Swine, an effective vaccine may be the best strategy to protect the pork industry and public health. Current inactivated-virus vaccines available for Swine Influenza protect only against viral strains closely related to the vaccine strain, and egg-based production of these vaccines is insufficient to respond to large outbreaks. DNA vaccines are a promising alternative since they can potentially induce broad-based protection with more efficient production methods. In this study we evaluated the potentials of monovalent and trivalent DNA vaccine constructs to (i) elicit both humoral and gamma interferon (IFN-γ) responses and (ii) protect pigs against viral shedding and lung disease after challenge with pandemic H1N1 or classic Swine H1N1 Influenza virus. We also compared the efficiency of a needle-free vaccine delivery method to that of a conventional needle/syringe injection. We report that DNA vaccination elicits robust serum antibody and cellular responses after three immunizations and confers significant protection against Influenza virus challenge. Needle-free delivery elicited improved antibody responses with the same efficiency as conventional injection and should be considered for development as a practical alternative for vaccine administration.
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efficacy of inactivated Swine Influenza virus vaccines against the 2009 a h1n1 Influenza virus in pigs
Vaccine, 2010Co-Authors: Amy L Vincent, Bruce H Janke, Eraldo L Zanella, Janice R Ciaccizanella, Marcus E Kehrli, Alessio Lorusso, P C Gauger, Kelly M LagerAbstract:The gene constellation of the 2009 pandemic A/H1N1 virus is a unique combination from Swine Influenza A viruses (SIV) of North American and Eurasian lineages, but prior to April 2009 had never before been identified in Swine or other species. Although its hemagglutinin gene is related to North American H1 SIV, it is unknown if vaccines currently used in U.S. Swine would cross-protect against infection with the pandemic A/H1N1. The objective of this study was to evaluate the efficacy of inactivated vaccines prepared with North American Swine Influenza viruses as well as an experimental homologous A/H1N1 vaccine to prevent infection and disease from 2009 pandemic A/H1N1. All vaccines tested provided partial protection ranging from reduction of pneumonia lesions to significant reduction in virus replication in the lung and nose. The multivalent vaccines demonstrated partial protection; however, none was able to prevent all nasal shedding or clinical disease. An experimental homologous 2009 A/H1N1 monovalent vaccine provided optimal protection with no virus detected from nose or lung at any time point in addition to amelioration of clinical disease. Based on cross-protection demonstrated with the vaccines evaluated in this study, the U.S. Swine herd likely has significant immunity to the 2009 A/H1N1 from prior vaccination or natural exposure. However, consideration should be given for development of monovalent homologous vaccines to best protect the Swine population thus limiting shedding and the potential transmission of 2009 A/H1N1 from pigs to people.
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experimental inoculation of pigs with pandemic h1n1 2009 virus and hi cross reactivity with contemporary Swine Influenza virus antisera
Influenza and Other Respiratory Viruses, 2010Co-Authors: Amy L Vincent, Kelly M Lager, Kay S Faaberg, Michelle Harland, Eraldo L Zanella, Janice R Ciaccizanella, Marcus E Kehrli, Bruce H JankeAbstract:Please cite this paper as: Vincent et al. (2010) Experimental inoculation of pigs with pandemic H1N1 2009 virus and HI cross-reactivity with contemporary Swine Influenza virus antisera. Influenza and Other Respiratory Viruses 4(2), 53–60 Background A novel A/H1N1 was identified in the human population in North America in April 2009. The gene constellation of the virus was a combination from Swine Influenza A viruses (SIV) of North American and Eurasian lineages that had never before been identified in Swine or other species. Objectives The objectives were to (i) evaluate the clinical response of Swine following experimental inoculation with pandemic H1N1 2009; (ii) assess serologic cross-reactivity between H1N1 2009 and contemporary SIV antisera; and (iii) develop a molecular assay to differentiate North American-lineage SIV from H1N1 2009. Methods Experiment 1: Weaned pigs were experimentally infected with A/California/04/2009 (H1N1). Experiment 2: The cross-reactivity of a panel of US SIV H1N1 or H1N2 antisera with three isolates of pandemic A/H1N1 was evaluated. Experiment 3: A polymerase chain reaction (PCR)-based diagnostic test was developed and validated on samples from experimentally infected pigs. Results and Conclusions In experiment 1, all inoculated pigs demonstrated clinical signs and lesions similar to those induced by endemic SIV. Viable virus and antigen were only detected in the respiratory tract. In experiment 2, serologic cross-reactivity was limited against H1N1 2009 isolates, notably among virus antisera from the same HA phylogenetic cluster. The limited cross-reactivity suggests North American pigs may not be fully protected against H1N1 2009 from previous exposure or vaccination and novel tests are needed to rapidly diagnose the introduction of H1N1 2009. In experiment 3, an RT–PCR test that discriminates between H1N1 2009 and endemic North American SIV was developed and validated on clinical samples.
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Swine Influenza matrix 2 m2 protein contributes to protection against infection with different h1 Swine Influenza virus siv isolates
Vaccine, 2009Co-Authors: Pravina Kitikoon, Amy L Vincent, Bruce H Janke, Marie Gramer, Barbara Z Erickson, Erin L Strait, Eileen L ThackerAbstract:A Swine Influenza virus (SIV) vaccine-challenge pig model was used to study the potential of a conserved matrix 2 (M2) protein vaccine alone or in combination with an inactivated H1N1-vaccine to protect against H1N1 and H1N2 viruses. The H1N1-vaccine and heterologous H1N2-challenge virus model has previously been shown to prolong fever and increase SIV-associated pneumonic lesions. The M2 vaccine in combination with the H1N1-vaccine reduced the H1N2 induced fever but not virus shedding. The M2 vaccine alone reduced respiratory signs and pneumonic lesions to levels similar to the negative control pigs following H1N2 infection. This study found that the M2 protein has potential as a vaccine for SIV-associated disease prevention. However, development of an immune response towards the major envelope HA protein was required to reduce SIV shedding.
Kelly M Lager - One of the best experts on this subject based on the ideXlab platform.
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dna vaccination elicits protective immune responses against pandemic and classic Swine Influenza viruses in pigs
Clinical and Vaccine Immunology, 2011Co-Authors: Patrick J Gorres, Kelly M Lager, Wing Pui Kong, Michael Royals, John Paul Todd, Amy L Vincent, Chih Jen Wei, Crystal L Loving, Eraldo Lourenso Zanella, Bruce H JankeAbstract:Swine Influenza is a highly contagious viral infection in pigs that significantly impacts the pork industry due to weight loss and secondary infections. There is also the potential of a significant threat to public health, as was seen in 2009 when the pandemic H1N1 Influenza virus strain emerged from reassortment events among avian, Swine, and human Influenza viruses within pigs. As classic and pandemic H1N1 strains now circulate in Swine, an effective vaccine may be the best strategy to protect the pork industry and public health. Current inactivated-virus vaccines available for Swine Influenza protect only against viral strains closely related to the vaccine strain, and egg-based production of these vaccines is insufficient to respond to large outbreaks. DNA vaccines are a promising alternative since they can potentially induce broad-based protection with more efficient production methods. In this study we evaluated the potentials of monovalent and trivalent DNA vaccine constructs to (i) elicit both humoral and gamma interferon (IFN-γ) responses and (ii) protect pigs against viral shedding and lung disease after challenge with pandemic H1N1 or classic Swine H1N1 Influenza virus. We also compared the efficiency of a needle-free vaccine delivery method to that of a conventional needle/syringe injection. We report that DNA vaccination elicits robust serum antibody and cellular responses after three immunizations and confers significant protection against Influenza virus challenge. Needle-free delivery elicited improved antibody responses with the same efficiency as conventional injection and should be considered for development as a practical alternative for vaccine administration.
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viral reassortment and transmission after co infection of pigs with classical h1n1 and triple reassortant h3n2 Swine Influenza viruses
Journal of General Virology, 2010Co-Authors: Kelly M Lager, Bruce H Janke, Adolfo Garciasastre, Jurgen A Richt, Porntippa Lekcharoensuk, Alicia Solorzano, Richard J Webby, Eva S UleryAbstract:Triple-reassortant Swine Influenza viruses circulating in North American pigs contain the internal genes derived from Swine (matrix, non-structural and nucleoprotein), human [polymerase basic 1 (PB1)] and avian (polymerase acidic and PB2) Influenza viruses forming a constellation of genes that is well conserved and is called the triple-reassortant internal gene (TRIG) cassette. In contrast, the external genes [haemagglutinin (HA) and neuraminidase (NA)] are less conserved, reflecting multiple reassortant events that have produced viruses with different combinations of HA and NA genes. This study hypothesized that maintenance of the TRIG cassette confers a selective advantage to the virus. To test this hypothesis, pigs were co-infected with the triple-reassortant H3N2 A/Swine/Texas/4199-2/98 (Tx/98) and the classical H1N1 A/Swine/Iowa/15/1930 viruses and co-housed with a group of sentinel animals. This direct contact group was subsequently moved into contact with a second group of naive animals. Four different subtypes (H1N1, H1N2, H3N1 and H3N2) of Influenza virus were identified in bronchoalveolar lavage fluid collected from the lungs of the experimentally infected pigs, with most of the viruses containing TRIG from the Tx/98 virus. Interestingly, only the intact H3N2 Tx/98 virus was transmitted from the infected pigs to the direct-contact animals and from them to the second contact group of pigs. These results demonstrated that multiple reassortments can occur within a host; however, only specific gene constellations are readily transmissible. It was concluded that certain HA and NA gene pairs, in conjunction with the TRIG cassette, may have a competitive advantage over other combinations for transmission and maintenance in Swine.
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efficacy of inactivated Swine Influenza virus vaccines against the 2009 a h1n1 Influenza virus in pigs
Vaccine, 2010Co-Authors: Amy L Vincent, Bruce H Janke, Eraldo L Zanella, Janice R Ciaccizanella, Marcus E Kehrli, Alessio Lorusso, P C Gauger, Kelly M LagerAbstract:The gene constellation of the 2009 pandemic A/H1N1 virus is a unique combination from Swine Influenza A viruses (SIV) of North American and Eurasian lineages, but prior to April 2009 had never before been identified in Swine or other species. Although its hemagglutinin gene is related to North American H1 SIV, it is unknown if vaccines currently used in U.S. Swine would cross-protect against infection with the pandemic A/H1N1. The objective of this study was to evaluate the efficacy of inactivated vaccines prepared with North American Swine Influenza viruses as well as an experimental homologous A/H1N1 vaccine to prevent infection and disease from 2009 pandemic A/H1N1. All vaccines tested provided partial protection ranging from reduction of pneumonia lesions to significant reduction in virus replication in the lung and nose. The multivalent vaccines demonstrated partial protection; however, none was able to prevent all nasal shedding or clinical disease. An experimental homologous 2009 A/H1N1 monovalent vaccine provided optimal protection with no virus detected from nose or lung at any time point in addition to amelioration of clinical disease. Based on cross-protection demonstrated with the vaccines evaluated in this study, the U.S. Swine herd likely has significant immunity to the 2009 A/H1N1 from prior vaccination or natural exposure. However, consideration should be given for development of monovalent homologous vaccines to best protect the Swine population thus limiting shedding and the potential transmission of 2009 A/H1N1 from pigs to people.
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experimental inoculation of pigs with pandemic h1n1 2009 virus and hi cross reactivity with contemporary Swine Influenza virus antisera
Influenza and Other Respiratory Viruses, 2010Co-Authors: Amy L Vincent, Kelly M Lager, Kay S Faaberg, Michelle Harland, Eraldo L Zanella, Janice R Ciaccizanella, Marcus E Kehrli, Bruce H JankeAbstract:Please cite this paper as: Vincent et al. (2010) Experimental inoculation of pigs with pandemic H1N1 2009 virus and HI cross-reactivity with contemporary Swine Influenza virus antisera. Influenza and Other Respiratory Viruses 4(2), 53–60 Background A novel A/H1N1 was identified in the human population in North America in April 2009. The gene constellation of the virus was a combination from Swine Influenza A viruses (SIV) of North American and Eurasian lineages that had never before been identified in Swine or other species. Objectives The objectives were to (i) evaluate the clinical response of Swine following experimental inoculation with pandemic H1N1 2009; (ii) assess serologic cross-reactivity between H1N1 2009 and contemporary SIV antisera; and (iii) develop a molecular assay to differentiate North American-lineage SIV from H1N1 2009. Methods Experiment 1: Weaned pigs were experimentally infected with A/California/04/2009 (H1N1). Experiment 2: The cross-reactivity of a panel of US SIV H1N1 or H1N2 antisera with three isolates of pandemic A/H1N1 was evaluated. Experiment 3: A polymerase chain reaction (PCR)-based diagnostic test was developed and validated on samples from experimentally infected pigs. Results and Conclusions In experiment 1, all inoculated pigs demonstrated clinical signs and lesions similar to those induced by endemic SIV. Viable virus and antigen were only detected in the respiratory tract. In experiment 2, serologic cross-reactivity was limited against H1N1 2009 isolates, notably among virus antisera from the same HA phylogenetic cluster. The limited cross-reactivity suggests North American pigs may not be fully protected against H1N1 2009 from previous exposure or vaccination and novel tests are needed to rapidly diagnose the introduction of H1N1 2009. In experiment 3, an RT–PCR test that discriminates between H1N1 2009 and endemic North American SIV was developed and validated on clinical samples.
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characterization of a newly emerged genetic cluster of h1n1 and h1n2 Swine Influenza virus in the united states
Virus Genes, 2009Co-Authors: Amy L Vincent, Kelly M Lager, Marie Gramer, Jurgen A Richt, Bruce H JankeAbstract:H1 Influenza A viruses that were distinct from the classical Swine H1 lineage were identified in pigs in Canada in 2003–2004; antigenic and genetic characterization identified the hemagglutinin (HA) as human H1 lineage. The viruses identified in Canadian pigs were human lineage in entirety or double (human–Swine) reassortants. Here, we report the whole genome sequence analysis of four human-like H1 viruses isolated from U.S. Swine in 2005 and 2007. All four isolates were characterized as triple reassortants with an internal gene constellation similar to contemporary U.S. Swine Influenza virus (SIV), with HA and neuraminidase (NA) most similar to human Influenza virus lineages. A 2007 human-like H1N1 was evaluated in a pathogenesis and transmission model and compared to a 2004 reassortant H1N1 SIV isolate with Swine lineage HA and NA. The 2007 isolate induced disease typical of Influenza virus and was transmitted to contact pigs; however, the kinetics and magnitude differed from the 2004 H1N1 SIV. This study indicates that the human-like H1 SIV can efficiently replicate and transmit in the Swine host and now co-circulates with contemporary SIVs as a distinct genetic cluster of H1 SIV.
Jurgen A Richt - One of the best experts on this subject based on the ideXlab platform.
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viral reassortment and transmission after co infection of pigs with classical h1n1 and triple reassortant h3n2 Swine Influenza viruses
Journal of General Virology, 2010Co-Authors: Kelly M Lager, Bruce H Janke, Adolfo Garciasastre, Jurgen A Richt, Porntippa Lekcharoensuk, Alicia Solorzano, Richard J Webby, Eva S UleryAbstract:Triple-reassortant Swine Influenza viruses circulating in North American pigs contain the internal genes derived from Swine (matrix, non-structural and nucleoprotein), human [polymerase basic 1 (PB1)] and avian (polymerase acidic and PB2) Influenza viruses forming a constellation of genes that is well conserved and is called the triple-reassortant internal gene (TRIG) cassette. In contrast, the external genes [haemagglutinin (HA) and neuraminidase (NA)] are less conserved, reflecting multiple reassortant events that have produced viruses with different combinations of HA and NA genes. This study hypothesized that maintenance of the TRIG cassette confers a selective advantage to the virus. To test this hypothesis, pigs were co-infected with the triple-reassortant H3N2 A/Swine/Texas/4199-2/98 (Tx/98) and the classical H1N1 A/Swine/Iowa/15/1930 viruses and co-housed with a group of sentinel animals. This direct contact group was subsequently moved into contact with a second group of naive animals. Four different subtypes (H1N1, H1N2, H3N1 and H3N2) of Influenza virus were identified in bronchoalveolar lavage fluid collected from the lungs of the experimentally infected pigs, with most of the viruses containing TRIG from the Tx/98 virus. Interestingly, only the intact H3N2 Tx/98 virus was transmitted from the infected pigs to the direct-contact animals and from them to the second contact group of pigs. These results demonstrated that multiple reassortments can occur within a host; however, only specific gene constellations are readily transmissible. It was concluded that certain HA and NA gene pairs, in conjunction with the TRIG cassette, may have a competitive advantage over other combinations for transmission and maintenance in Swine.
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characterization of a newly emerged genetic cluster of h1n1 and h1n2 Swine Influenza virus in the united states
Virus Genes, 2009Co-Authors: Amy L Vincent, Kelly M Lager, Marie Gramer, Jurgen A Richt, Bruce H JankeAbstract:H1 Influenza A viruses that were distinct from the classical Swine H1 lineage were identified in pigs in Canada in 2003–2004; antigenic and genetic characterization identified the hemagglutinin (HA) as human H1 lineage. The viruses identified in Canadian pigs were human lineage in entirety or double (human–Swine) reassortants. Here, we report the whole genome sequence analysis of four human-like H1 viruses isolated from U.S. Swine in 2005 and 2007. All four isolates were characterized as triple reassortants with an internal gene constellation similar to contemporary U.S. Swine Influenza virus (SIV), with HA and neuraminidase (NA) most similar to human Influenza virus lineages. A 2007 human-like H1N1 was evaluated in a pathogenesis and transmission model and compared to a 2004 reassortant H1N1 SIV isolate with Swine lineage HA and NA. The 2007 isolate induced disease typical of Influenza virus and was transmitted to contact pigs; however, the kinetics and magnitude differed from the 2004 H1N1 SIV. This study indicates that the human-like H1 SIV can efficiently replicate and transmit in the Swine host and now co-circulates with contemporary SIVs as a distinct genetic cluster of H1 SIV.
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failure of protection and enhanced pneumonia with a us h1n2 Swine Influenza virus in pigs vaccinated with an inactivated classical Swine h1n1 vaccine
Veterinary Microbiology, 2008Co-Authors: Amy L Vincent, Kelly M Lager, Bruce H Janke, Marie Gramer, Jurgen A RichtAbstract:Two US Swine Influenza virus (SIV) isolates, A/Swine/Iowa/15/1930 H1N1 (IA30) and A/Swine/Minnesota/00194/2003 H1N2 (MN03), were evaluated in an in vivo vaccination and challenge model. Inactivated vaccines were prepared from each isolate and used to immunize conventional pigs, followed by challenge with homologous or heterologous virus. Both inactivated vaccines provided complete protection against homologous challenge. However, the IA30 vaccine failed to protect against the heterologous MN03 challenge. Three of the nine pigs in this group had substantially greater percentages of lung lesions, suggesting the vaccine potentiated the pneumonia. In contrast, priming with live IA30 virus provided protection from nasal shedding and virus replication in the lung in MN03 challenged pigs. These data indicate that divergent viruses that did not cross-react serologically did not provide complete cross-protection when used in inactivated vaccines against heterologous challenge and may have enhanced disease. In addition, live virus infection conferred protection against heterologous challenge.
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evaluation of hemagglutinin subtype 1 Swine Influenza viruses from the united states
Veterinary Microbiology, 2006Co-Authors: Amy L Vincent, Kelly M Lager, Marie Gramer, Porntippa Lekcharoensuk, Christina M Loiacono, Jurgen A RichtAbstract:Swine Influenza viruses (SIV) of the hemagglutinin subtype 1 (H1) isolated from the United States (U.S.) have not been well-characterized in the natural host. An increase in the rate of mutation and reassortment has occurred in SIV isolates from the U.S. since 1998, including viruses belonging to the H1 subtype. Two independent animal studies were done to evaluate and compare the pathogenesis of 10 SIV isolates dating from 1930 to currently circulating isolates. In addition, the hemagglutinin and neuraminidase genes of each isolate were sequenced for genetic comparison, and serological cross-reactivity was evaluated using all sera and virus combinations in hemagglutination inhibition and serum neutralization assays. Statistically significant differences in percentage of pneumonia and virus titers in the lung were detected between isolates, with modern isolates tending to produce more severe disease, have more virus shedding and higher viral titers. However, nasal shedding and virus titers in the lung were not always correlated with one another or lung lesions. Serologically, the classic historical H1N1 viruses tended to have better cross-reaction between historical sera and antigens, with moderate to good cross-reactivity with modern viral antigens. However, the modern sera were less reactive with historical viruses. Modern viruses tended to have less consistent cross-reactivity within the modern group. Overall, H1 isolates collected over the last 75 years from the U.S. pig population exhibit considerable variability in pathogenicity. There appears to be an increase in genetic and antigenic diversity coincident with the emergence of the Swine triple reassortant H3N2 in 1998.
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vaccination of pigs against Swine Influenza viruses by using an ns1 truncated modified live virus vaccine
Journal of Virology, 2006Co-Authors: Jurgen A Richt, Kelly M Lager, Amy L Vincent, Bruce H Janke, Kyoungjin Yoon, Porntippa Lekcharoensuk, Alicia Solorzano, Richard J Webby, Christina M Loiacono, Adolfo GarciasastreAbstract:Swine Influenza viruses (SIV) naturally infect pigs and can be transmitted to humans. In the pig, genetic reassortment to create novel Influenza subtypes by mixing avian, human, and Swine Influenza viruses is possible. An SIV vaccine inducing cross-protective immunity between different subtypes and strains circulating in pigs is highly desirable. Previously, we have shown that an H3N2 SIV (A/Swine/Texas/4199-2/98 [TX98]) containing a deleted NS1 gene expressing a truncated NS1 protein of 126 amino acids, NS1Œ126, was attenuated in Swine. In this study, 4-week-old pigs were vaccinated with the TX98 NS1Œ126 modified live virus (MLV). Ten days after boosting, pigs were challenged with wild-type homologous H3N2 or heterosubtypic H1N1 SIV and sacrificed 5 days later. The MLV was highly attenuated and completely protected against challenge with the homologous virus. Vaccinated pigs challenged with the heterosubtypic H1N1 virus demonstrated macroscopic lung lesions similar to those of the unvaccinated H1N1 control pigs. Remarkably, vaccinated pigs challenged with the H1N1 SIV had significantly lower microscopic lung lesions and less virus shedding from the respiratory tract than did unvaccinated, H1N1-challenged pigs. All vaccinated pigs developed significant levels of hemagglutination inhibition and enzyme-linked immunosorbent assay titers in serum and mucosal immunoglobulin A antibodies against H3N2 SIV antigens. Vaccinated pigs were seronegative for NS1, indicating the potential use of the TX98 NS1Œ126 MLV as a vaccine to differentiate infected from vaccinated animals. Swine Influenza virus (SIV) is an important Swine pathogen involved in the porcine respiratory disease complex in most Swine-producing countries. Serological and virological studies demonstrated 23 to 28% prevalence of SIV in Swine populations in the midwestern and north central United States (4, 35, 56). Mortality of SIV in pigs is low, while morbidity may approach 100%. SIV-affected pigs show clinical signs of anorexia and weight loss, fever, respiratory distress, coughing, and nasal discharge (18). SIVs currently circulating in North American Swine are subtypes H1N1, H3N2, and H1N2 (36). From 1930 to 1998, classical H1N1 viruses were the subtype predominantly isolated from U.S. Swine. In 1998, a new SIV subtype, H3N2, emerged from reassortment of Swine, human, and avian Influenza viruses (19, 56, 58). The H3N2 SIV acquired PB1, hemagglutinin (HA), and NA genes from recent human viruses, PB2 and PA from avian viruses, and NP, M, and NS genes from the classical H1N1 Swine virus (19, 56, 58). A year
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Identification of a Newly Conserved SLA-II Epitope in a Structural Protein of Swine Influenza Virus.
Frontiers in immunology, 2020Co-Authors: Massimiliano Baratelli, Johanneke D. Hemmink, Bryan Charleston, Sophie B. Morgan, Elizabeth Reid, Brigid Veronica Carr, Eric A. Lefevre, Sergio Montaner-tarbes, Lorenzo Fraile, Elma TchilianAbstract:Despite the role of pigs as a source of new Influenza A Virus viruses (IAV) potentially capable of initiating human pandemics, immune responses to Swine Influenza virus (SwIV) in pigs are not fully understood. Several SwIV epitopes presented by Swine MHC (SLA) class I have been identified using different approaches either in outbred pigs or in Babraham large white inbred pigs, which are 85% identical by genome wide SNP analysis. On the other hand, some class II SLA epitopes were recently described in outbred pigs. In this work, Babraham large white inbred pigs were selected to identify SLA II epitopes from SwIV H1N1. PBMCs were screened for recognition of overlapping peptides covering the NP and M1 proteins from heterologous IAV H1N1 in IFNγ ELISPOT. A novel SLA class II restricted epitope was identified in NP from Swine H1N1. This conserved novel epitope could be the base for further vaccine approaches against H1N1 in pigs.
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vaccine mediated protection of pigs against infection with pandemic h1n1 2009 Swine Influenza a virus requires a close antigenic match between the vaccine antigen and challenge virus
Vaccine, 2019Co-Authors: Helen Everett, Elma Tchilian, Mario Aramouni, Andrew Ramsay, Vivien J Coward, Laetitia Canini, Michael Kelly, Sophie Morgan, Mark E J Woolhouse, Sarah C GilbertAbstract:Swine Influenza A virus (SwIV) infection has considerable economic and animal welfare consequences and, because of the zoonotic potential, can also have public health implications. The 2009 pandemic H1N1 'Swine-origin' infection is now endemic in both pigs and humans. In Europe, avian-like H1avN1, human-like H1huN2, human-like Swine H3N2 and, since 2009, pandemic H1N1 (pH1N1) lineage viruses and reassortants, constitute the dominant subtypes. In this study, we used a Swine pH1N1 challenge virus to investigate the efficacy of whole inactivated virus vaccines homologous or heterologous to the challenge virus as well as a commercial vaccine. We found that vaccine-mediated protection was most effective when vaccine antigen and challenge virus were homologous and correlated with the specific production of neutralising antibodies and a cellular response to the challenge virus. We conclude that a conventional whole inactivated SwIV vaccine must be antigenically matched to the challenge strain to be an effective control measure.
