Source: UNIVERSITY OF GEORGIA submitted to NRP
BROADLY PROTECTIVE MODIFIED LIVE ATTENUATED INFLUENZA VACCINES FOR POULTRY
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
AFRI COMPETITIVE GRANT
Reporting Frequency
Annual
Accession No.
1022658
Grant No.
2020-67015-31539
Cumulative Award Amt.
$500,000.00
Proposal No.
2019-05890
Multistate No.
(N/A)
Project Start Date
Jul 1, 2020
Project End Date
Jun 30, 2024
Grant Year
2020
Program Code
[A1221]- Animal Health and Production and Animal Products: Animal Health and Disease
Recipient Organization
UNIVERSITY OF GEORGIA
200 D.W. BROOKS DR
ATHENS,GA 30602-5016
Performing Department
(N/A)
Non Technical Summary
Avian influenza virus (AIV) is a devastating diseasewith demonstrated ability to cause the loss of hundreds of millions of birds.The major goal of this project is totest the hypothesis that modified live virus (MLV) vaccinesinduce greater and more broadly reactive immune responses and provide superior protection than standard inactivated vaccines against avian influenza.The poultry industry, comprising of billions of birds, is of huge economic and social importance throughout the world. In some countries, vaccination has been introduced to prevent AIV outbreaks; however,current vaccines offerlimited protection and cannot prevent the spread of the virus. Nowadays, the world confronts anunprecedented spread ofAIVs in poultry inAsia, the Middle East and Africa, with frequent spill over andtranscontinental spread of these viruses through migratory birds to other parts of the world, including the U.S. Thus, it is absolutely essentialto better understandthe mechanisms involved in vaccine immunogenicity and todevelopalternative vaccination strategies that can more effectively contribute to prevent and control of AIV.We are a strong collaborative team with expertise in influenza virus reverse genetics, avian immunology and avian models of influenza disease to develop MLV vaccines againstAIVs for use in poultry. We will compare the characteristics of MLV vaccinesin vitroandin vivoand examine safety,transmission potential of the vaccines to susceptible birds, and genetic variability of challenge virusout of vaccinated versus non-vaccinatedbirds.
Animal Health Component
65%
Research Effort Categories
Basic
35%
Applied
65%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
3113999109050%
3113999104050%
Knowledge Area
311 - Animal Diseases;

Subject Of Investigation
3999 - Animal research, general;

Field Of Science
1040 - Molecular biology; 1090 - Immunology;
Goals / Objectives
The major goal of this application is to generate a novel class of modified live virus vaccines against influenza that elicit broadly protective immune responses.Avian influenza virus (AIV) is a devastating diseasewith demonstrated ability to cause the loss of hundreds of millions of birds.Control strategies for poultry against AIV is dependent on biosecurity to keep the virus out and vaccine application when the virus is present.Vaccine protection is principally mediated by an immune response to the subtype-specific hemagglutinin protein (HA) with inactivated virus vaccines the overwhelming choice in the field. However, this type of vaccine has a narrow range of activity that is not long lasting. In addition, not all HA subtypes of AIV are as immunogenic as others. It is widely known that live virus vaccines induce the greatest immune responses, but concerns of reversion, recombination and unintended transmission have slowed their development in poultry. Reverse genetics has allowed greater control of the viral genome which has helped minimize the perceived potential risks associated with live influenza virus vaccines; however, the mechanisms necessary to improve AIV vaccines with broader immunogenicity and increased suppression of virus transmission require further studies. The major goal of this application is to close such knowledge gap to ultimately improve the effectiveness of control programs for AIV and decrease its economic and public health burden. Since the H5 and H9 subtypes are two most widely spread AIVs in poultry around the world, we propose the following specific aims:Specific Aim 1:To generate and compare alternative modified live avian influenza virus vaccine platforms as vaccines against H5, and H9 subtypes of avian influenza viruses.Specific Aim 2:Test safety and protection of poultry against HPAI and LPAI challenge following MLV vaccination.Specific Aim 3:To characterize the impact of live virus vaccination on genomic mutation by deep sequencing analysis of virus shed from vaccinated birds.The application maximizes our current understanding of vaccines and immune responses against influenza in poultry and reverse genetics to produce MLV-AI vaccines with unique and novel set of features. These features are built-inin the context of high yield strains to potentiate protective mucosal responses while serving as molecular diagnostic markers that also reduce transmission and reassortment of the vaccine strains. If successful, our approach can be implemented in vaccine strategies against not only other AIV subtypes but also other viral respiratory pathogens of poultry and livestock.
Project Methods
We have taken into consideration the concerns of stakeholders and regulatory agencies and established a set ofparameters for an ideal MLV-AI vaccine to: 1) preserve the in vitro growth characteristics of the MLV vaccine similar to wild type levels, 2) maximize mucosal immune responses, 3) incorporate unique molecular diagnostic markers, 4) increase the breath of the immune response, and 5) prevent transmission and reassortment. We propose to design these features and compare two different MLV-AI vaccine strategies (MLV-att and RAM). In addition,we propose to modify candidate H5 and H9 HA segments to carry molecular diagnostic markers as well as modifications to enhance mucosal responses.Recombinant virus characterization will be performed by analyzing in vitro growth kinetics at different temperatures in tissue culture cells, virus yields in SPF eggs, stability over at least 5 serial passages in both tissue culture cells and eggs and finally sequencing by next generation sequencing technologies and a bioinformatics pipeline developed in-house. The completion of aim 1will produceMLV-att and RAM vaccine candidates with M2 ORF deletions and with genetic markers and modifications that reduce fitness for reassortment and enhance mucosal responses. In aim 2, we will provideunequivocal information regarding the immunogenicity and replication and transmission potentialof the MLV vaccines as well astheir efficacyin poultry. Since commercial poultry farms and live bird markets include both chickens and ducks, both will be used for the in vivo characterization of the MLV-AI vaccine viruses. Comparison of the pathogenicity (morbidity and mortality), virus replication, tissue tropism and immune response to the MLV vaccines will be examined. And finally in aim 3, we will establish whether birds vaccinated with MLV-AIs leads to either faster, slower or neutral rates of mutations on the challenge virus compared to similar viruses obtained from birds vaccinated with inactivated vaccines.These studies will not only reveal the extent of mutations in the challenge viruses but also it will generate highly significant information regarding the evolutionary path taken by the challenge viruses exposed to different immunological environments (MLV-AI vs inactivated vs non-vaccinated). We expect to have reduced shedding of the challenge virus in birds vaccinated with MLV-AIs compared to those vaccinated with commercially available inactivated vaccines.Numbers of birds used for experiments are determined by previous studiesand are based on sample sets sufficient to demonstrate statistical significance. According to OIE and USDA regulations, pathotyping of avian viruses, derivatives or reassortants requires the use of 8 birds, 4-6 weeks of age, per test. We will add additional 2-3 birds to each test for sampling of tissues for histopathology, immunohistochemistry, virus isolation and other tests that require termination of birds and must be excluded from the official pathotyping tests. The number of birds to be examined per time point were determined based on statistical power analysis in order to provide a Power of at least 0.8 with α = 0.05. The data collected in our previous studies was used in the power analysis calculation. Data will be analyzed by ANOVA. Treatment means will be separated by Tukeys multiple comparison test. All data will be evaluated for approximately normal distribution and similar group variance. Transformations, such as log10, will be used to correct for skewness. We will use longitudinal analyses to explore the relationships between groups and outcomes over all time points.The application maximizes our current understanding of vaccines and immune responses against influenza in poultry and reverse genetics to produce MLV-AI vaccines with unique and novel set of features.If successful, our approach can be implemented in vaccine strategies against not only other AIV subtypes but also other viral respiratory pathogens of poultry and livestock.

Progress 07/01/20 to 05/31/24

Outputs
Target Audience:Poultry industry, veterinary vaccine companies, academic research centers, one health experts. Changes/Problems:The grant started during the COVID-19 pandemic that affected the overall goals of the application, particularly in terms of further developing vaccines against the H5 subtype influenza viruses. Despite this major roadblock, we were productive and achieved most of the goals of the application regarding the development of modified live virus vaccines agains the H9 subtype. What opportunities for training and professional development has the project provided?The project has provided plenty of training opportunities in molecular biology and reverse genetics to post-doctoral fellows (C. Joaquin Caceres, Juliana Brondani, graduate students (Ginger Geiger, Lindsay Gay, Flavio Faccin, Matias Cardenas, Teresa Mejias, Silvia Carnaccini, Dikshya Regmi) and two UGA Post-Bacculaureate Research Program (PREP) Scholars (Luis Rodriguez and Blake Vilchez). How have the results been disseminated to communities of interest?My personnel and I have had the opportunity to present relevant data in a myriad of national and international venues. Listed below those the personnel from my lab or collaborators have presented. Neveau, M., Cowan, B., Caceres, C. J., de Souza Rajao, D., Perez, D., Gauger, P., . . . Anderson, T. (2023). The effect of nucleoprotein reassortment on influenza A virus transmission in swine. American Association of Swine Veterinarians. Cardenas, M., Cowan, B., Seibert, B., Gay, L., Cargnin, F., Caceres, C. J., . . . de Souza Rajao, D. (2023). Multiple transmissions of human influenza A virus in pigs leads to improved replication. Conference of Research Workers in Animal Diseases (CRWAD2023). Cargnin, F., Caceres, C. J., Gay, L., Van Bentem, N., Seibert, B., Rodriguez, L., . . . Perez, D. (2023). Development of a modified live attenuated influenza virus vaccine against H9N2 for poultry. Conference of Research Workers in Animal Diseases (CRWAD) 2023. Caceres, C. J., Carnaccini, S., Gay, L. C., Ferreri, L., Skepner, E., Burke, D., . . . Perez, D. (2023). Antigenic map of the hemagglutinin of influenza A virus of the H9 subtype. Conference of Research Workers in Animal Diseases (CRWAD) 2023. Caceres, C., Cargnin Faccin, F., Gay, L., Jain, A., Seibert, B., Rodriguez, L. A., . . . Perez, D. (2022). Genome rearrangement for live attenuated influenza virus vaccines. XLIV Annual meeting of the Chilean Society of Microbiology, La Serena, Chile. Gay, L., Caceres, C., Seibert, B., Cargnin Faccin, F., Graziosi, G., Cowan, B., . . . Perez, D. R. (2022). Impact of sex and virus lineage on Influenza B virus pathogenesis in the DBA/2J mice model.. Steeve Giguere Science of Veterinary Medicine Symposium, University of Georgia, Athens, GA.. Cargnin Faccin, F., Caceres, C., Gay, L., van Bentem, N., Seibert, B., Rodriguez, L., . . . Perez, D. (2022). Development of a modified live attenuated influenza virus vaccine against H9N2.. Steeve Giguere Science of Veterinary Medicine Symposium. Cargnin Faccin, F., Cáceres, C. J., Gay, L., van Bentem, N., Seibert, B., Rodriguez, L., . . . Perez, D. (2022). Development of a modified live attenuated influenza virus vaccine against H9N2.. XVI Avian Immunology Research Group Meeting. Newark, DE.. Gay, L., Caceres, C., Seibert, B., Cargnin Faccin, F., Graziosi, G., Cowan, B. L., . . . Perez, D. R. (2022). Impact of sex and virus lineage on Influenza B virus pathogenesis in the DBA/2J mice model.. Center of Excellence for Influenza Research and Response (CEIRR). Memphis, TN.. Caceres, C., Carnaccini, S., Gay, L., Ferreri, L., Skepner, E., Burke, D., . . . Perez, D. R. (2022). Antigenic map of the hemagglutinin of influenza A virus of the H9 subtype.. Annual meeting Center of Excellence for Influenza Research and Response (CEIRR). Memphis, TN.. Seibert, B., Caceres, C., Carnaccini, S., Cardenas-Garcia, S., Gay, L., Ortiz, L., . . . Perez, D. (2022). The effect of SARS-CoV-2 virus infection on the respiratory and intestinal microbiome in aged Golden Syrian hamsters.. American Society for Virology (ASV). Madison, WI.. Caceres, C., Cargnin Faccin, F., Rodriguez, L., Cardenas-Garcia, S., Gay, L., Ortiz, L., . . . Perez, D. (2022). Genome rearrangement for live attenuated influenza virus vaccines.. 41th Annual meeting of the American Society for Virology (ASV). Madison, WI.. Seibert, B., Caceres, C., Carnaccini, S., Cardenas-Garcia, S., Gay, L., Ortiz, L., . . . Perez, D. (2022). Pathobiology and dysbiosis of the respiratory and intestinal microbiota in aged Golden Syrian hamsters infected with SARS-CoV-2. Centers of Excellence for Influenza Research and Response (CEIRR). Memphis, TN. What do you plan to do during the next reporting period to accomplish the goals?This is the final report. The work continues under the support a new NIFA application.

Impacts
What was accomplished under these goals? H9N2 antigenic make up (Carnaccini et al, JVI 2023): Current research on the antigenic properties of H9N2 avian influenza viruses (AIVs) is limited in scope, focusing primarily on specific regions and using chicken sera. This study aimed to expand this understanding by including a minor poultry species, Japanese quail, and analyzing a global set of H9N2 viruses. Key findings include: Identification of 4 distinct antigenic clusters of H9N2 viruses. Determination of 8-9 amino acid positions in the H9 hemagglutinin (HA) protein that significantly influence antigenicity. Observation that antigenic maps generated using chicken and quail sera may differ, suggesting species-specific recognition of H9N2 viruses. Development of a standardized method for producing anti-H9 sera in both chickens and quail, enabling more accurate assessment of antigenic drift. These findings provide a more comprehensive understanding of the antigenic properties of H9N2 LPAIVs and lay the foundation for developing more effective and broadly protective vaccines. The identification of key amino acid positions involved in antigenic drift can inform the design of vaccines that target these regions, potentially offering broader cross-protection against diverse H9N2 strains. Mass vaccination against H9N2 avian influenza A virus with a non-transmissible, reassortment-impaired modified live attenuated influenza virus vaccine(Faccin et al, revision under consideration in npjVaccines) Research Paper highlights:The prevalent H9N2 avian influenza virus (AIV) poses a significant threat to the poultry industry and public health.Current inactivated vaccines lack efficacy in preventing virus shedding and transmission. Modified live virus (MLV) vaccines, on the other hand, offer a promising alternative due to their ability to mimic natural infection and induce broader immune responses. This study developed two MLVs, MLV-H9N2 and MLV-H9N2-IL, utilizing a genome rearrangement approach and incorporating molecular markers to prevent reassortment. The MLVs were genetically stable, attenuated in vivo, and did not reassort with wild-type viruses. Notably, MLV-H9N2-IL, containing an immunomodulator, demonstrated superior protection compared to the inactivated vaccine and the MLV without IL-18. Both MLVs effectively stimulated humoral immune responses and reduced viral shedding in chickens. MLV-H9N2-IL,when administered via drinking water, induced sterilizing immunity in some birds, demonstrating its potential for mass vaccination and controlling the transmission cycle. Although the current MLVs do not support a Differentiating Infected from Vaccinated Animals (DIVA) strategy, the focus on non-notifiable H9N2 virus and the lack of widespread DIVA implementation in many countries render this less critical.Future research will explore further modifications to the MLVs that could enable DIVA implementation and enhance their applicability in diverse settings. Overall, this study highlights the potential of MLVs as effective vaccines against avian influenza. The inclusion of molecular markers and immunomodulators like IL-18 demonstrates the flexibility and potential of this platform for further optimization and development of broadly protective vaccines. Pandemic preparedness through vaccine development for avian influenza viruses(Faccin and Perez,HUMAN VACCINES & IMMUNOTHERAPEUTICS, 2024). Review highlights: Avian influenza viruses, particularly the H9N2, H5N1, and H7N9 subtypes, present a significant challenge to both animal health and human health. The economic impact on the poultry industry is substantial, and the risk of zoonotic transmission and subsequent pandemics necessitates robust vaccination strategies. Traditional vaccine platforms, including inactivated whole-virus adjuvanted vaccines and live attenuated vaccines, have been extensively studied in animal models and humans. Inactivated vaccines offer safety and cost-effectiveness but primarily induce humoral immunity. Live attenuated vaccines, while potentially more protective due to their broader immune response (humoral, mucosal, and cell-mediated), raise concerns about safety and manufacturing complexities. The development of novel vaccine platforms, such as virus-like particles (VLPs) and mRNA-lipid nanoparticle (LNP) vaccines, offers promising alternatives. VLPs mimic the native virus structure without containing genetic material,ensuring safety while eliciting potent immune responses. mRNA-LNP vaccines, as demonstrated in the COVID-19 pandemic, have the potential for rapid development and production, making them attractive for pandemic preparedness. A diversified vaccine portfolio is essential to effectively combat the evolving threat of avian influenza viruses. Continued research and development of various vaccine platforms will enhance our ability to respond to potential outbreaks and safeguard both animal and human populations.

Publications

  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Carnaccini, S., C�ceres, C. J., Gay, L. C., Ferreri, L. M., Skepner, E., Burke, D. F., . . . Perez, D. R. (2023). Antigenic mapping of the hemagglutinin of the H9 subtype influenza A viruses using sera from Japanese quail (Coturnix c. japonica). Journal of Virology, 97(10). doi:10.1128/jvi.00743-23
  • Type: Journal Articles Status: Under Review Year Published: 2024 Citation: Mass vaccination against H9N2 avian influenza A virus with a non-transmissible, reassortment-impaired modified live attenuated influenza virus vaccine. Flavio Cargnin Faccin, C. Joaquin C�ceres, L. Claire Gay, Brittany Seibert, Nick van Bentem, Luis A. Rodriguez, Ana Luiza Soares Fraiha, Matias Cardenas, Ginger Geiger, Lucia Ortiz, Silvia Carnaccini, Darrell R. Kapczynski, Daniela S. Rajao, and Daniel R. Perez. revision submitted to npjVaccines
  • Type: Other Status: Published Year Published: 2024 Citation: Perez, D. R. (2024). A devastating blow: personal reflections on Argentinas scientific decline. Journal of Virology, 98(5). doi:10.1128/jvi.00549-24
  • Type: Journal Articles Status: Published Year Published: 2024 Citation: Thomas, M. N., Zanella, G. C., Cowan, B., Caceres, C. J., Rajao, D. S., Perez, D. R., . . . Anderson, T. K. (2024). Nucleoprotein reassortment enhanced transmissibility of H3 1990.4.a clade influenza A virus in swine. Journal of Virology, 98(3). doi:10.1128/jvi.01703-23
  • Type: Journal Articles Status: Published Year Published: 2024 Citation: Curran, S. J., Griffin, E. F., Ferreri, L. M., Kyriakis, C. S., Howerth, E. W., Perez, D. R., & Tompkins, S. M. (2024). Swine influenza A virus isolates containing the pandemic H1N1 origin matrix gene elicit greater disease in the murine model. Microbiology Spectrum, 12(3). doi:10.1128/spectrum.03386-23
  • Type: Journal Articles Status: Published Year Published: 2024 Citation: Cardenas-Garcia, S., C�ceres, C. J., Jain, A., Geiger, G., Mo, J. -S., Gay, L. C., . . . Perez, D. R. Impact of sex on humoral immunity with live influenza B virus vaccines in mice. npj Vaccines, 9(1). doi:10.1038/s41541-024-00827-x
  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Cardenas, M., Seibert, B., Cowan, B., Fraiha, A. L. S., Carnaccini, S., Gay, L. C., . . . Rajao, D. S. (n.d.). Amino acid 138 in the HA of a H3N2 subtype influenza A virus increases affinity for the lower respiratory tract and alveolar macrophages in pigs. PLOS Pathogens, 20(2), e1012026. doi:10.1371/journal.ppat.1012026
  • Type: Journal Articles Status: Under Review Year Published: 2024 Citation: Joaqu�n C�ceres, C., Claire Gay, L., Jain, A., Mej�as, T. D., Cardenas, M., Seibert, B., . . . Perez, D. R. (2024). FLUAV RAM-IGIP: A modified live influenza virus vaccine that enhances humoral and mucosal responses against influenza. doi:10.1101/2024.01.23.576908


Progress 07/01/22 to 06/30/23

Outputs
Target Audience:Poultry interest groups, One health groups Changes/Problems:Circumtances beyond everyone's control have prevented our collaborators at USDA from moving forward with the vaccination/challenge studies that were planned for the H5 subtype influenza viruses. What opportunities for training and professional development has the project provided?Two graduate students and two postdocs have been working on this project How have the results been disseminated to communities of interest?We have presented results during the annual meeting of the American Society for Virology (2023), the American Association of Avian Pathologists (2023), and the International Society for Vaccines (2023). What do you plan to do during the next reporting period to accomplish the goals?We are moving right along with the project. A manuscript is being prepared and a grant application to NIFA is being prepared.

Impacts
What was accomplished under these goals? MLV with immunomodulators were produced against H9 and H5 subtype influenza viruses. Only the H9 vaccine candidates were teste. We have had remarkable results in terms of safety and protection while exploring mass vaccine application through drinking water.

Publications


    Progress 07/01/21 to 06/30/22

    Outputs
    Target Audience:Avian influenza virus (AIV) is a devastating disease with demonstrated ability to cause the loss of hundreds of millions of birds. The major goal of this project is to test the hypothesis that modified live virus (MLV) vaccines induce greater and more broadly reactive immune responses and provide superior protection than standard inactivated vaccines against avian influenza. The poultry industry, comprising of billions of birds, is of huge economic and social importance throughout the world. The current panzootic caused by H5N1 HPAIV further highlights the urgent need to develop alternative vaccination strategies to prevent and control avian influenza in birds. Over the past year, we have made significant progress in our project, particularly in the development of vaccines against low pathogenic Eurasian-origin H9N2 avian influenza viruses, which continue to cause significant outbreaks in poultry. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The project has allowed training of two graduate students and three post-docs that have learned, classical virology, reverse genetics, animal models of influenza and immunology How have the results been disseminated to communities of interest?At least one manuscript for peer-review publication will be produced during 2023. What do you plan to do during the next reporting period to accomplish the goals?We will continue as planned, particularly expanding on similar approaches to vaccinate birds against HPAIV H5N1 strains.

    Impacts
    What was accomplished under these goals? Why MLVs? Because they can provide multidimensional mucosal, humoral and cell-mediated responses - Mimic live virus infection - Mass vaccination. RAM-approach (MLV-H9N2 and MLV-H9N2-IL) - Rearranged M segment PB1 + M2 RAM42: PB1 + M42 Early stop codons in original M2. Flu-UNIQUE (MLV-H9N2 and MLV-H9N2-IL) Molecular marker in segment 4 (HA). Diagnostic tool and reduced fitness. Flu-kine (MLV-H9N2-IL) Immunomodulator introduced in segment 6 (NA) Stimulate mucosal responses. MLVs underwent five serial passages in eggs. We confirmed that the rearranged genomes are stable after five serial passages in eggs (E5) No mutations were found in the PB1, HA, NA, and M segments of rearranged viruses. Next-Generation-Sequencing (NGS) is ongoing. MLVs show similar growth profiles in comparison to prototypic wild-type H9N2 strain. Coinfections in vitro show HA/NA segment fitness cost associated with modifications. In vivo studies: Hemagglutination Inhibition (HI) and Virus Neutralization assays (VNLuc) were performed to detect neutralizing antibodies. NP ELISA was performed to monitor vaccination/infection. HI and VN neutralizing antibodies are readily detected after prime using MLVs. HI titers consistently higher with WIV-adj vaccine. The introduction of the immunomodulator leads to higher titers after prime and boost. NP antibodies consistent with MLV replication. HI titers show limited cross-reactivity among clades. WIV-adj consistently higher HI despite limited cross-reactivity. MLV with immunomodulator significantly reduces virus load and enhances protection after challenge. The inactivated-adjuvanted vaccine provides limited protection for virus shedding. The introduction of the immunomodulator "helps" with cross-reactivity. MLVs significantly reduce viral load in the respiratory tract and cloaca. Lungs and PANCREAS: MLVs significantly reduce viral load compared to WIV-adj. vaccine. MLV-H9N2-IL is stable during drinking water vaccine administration. Serological response (HI, NP ELISA) can be detected 12 days after drinking water vaccination - Implies mucosal immunity? Limited replication of challenge virus in the respiratory and intestinal tracts of chickens after prime/boost with MLV-H9N2-IL in drinking H2O. ?

    Publications

    • Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: Sub types Article Title Antigenic map of the hemagglutinin of influenza A virus of the H9 subtype Authors Caceres, Carlos Joaquin; Carnaccini, Silvia; Gay, L. Claire; Ferreri, Lucas; Skepner, Eugene; Burke, David; Brown, Ian; Cardenas-Garcia, Stivalis; Geiger, Ginger; Obadan, Adebimpe, et al. Status Published Publication/Status date 30 Jan 2023 Journal Conference of Research Workers in Animal Diseases (CRWAD) 2023
    • Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: Sub types Rapid Communication, Article Title Genome rearrangement for live attenuated influenza virus vaccines Authors Caceres, Carlos; Cargnin Faccin, Flavio; Gay, L.; Jain, A.; Seibert, Brittany; Rodriguez, L. A.; Cowan, B.; Cardenas, M.; Van Bentem, N.; Geiger, G., et al. Status Published Publication/Status date 28 Nov 2022 Journal XLIV Annual meeting of the Chilean Society of Microbiology, La Serena, Chile
    • Type: Conference Papers and Presentations Status: Accepted Year Published: 2023 Citation: Sub types Article Title Development of a modified live attenuated influenza virus vaccine against H9N2 for poultry Authors Cargnin, Flavio; Caceres, Carlos Joaquin; Gay, Lindsey; Van Bentem, Nick; Seibert, Brittany; Rodriguez, Luis; Geiger, Ginger; Cowan, Brianna; Cardenas, Matias; de Souza Rajao, Daniela, et al. Status Accepted Publication/Status date 30 Jan 2023 Journal Conference of Research Workers in Animal Diseases (CRWAD) 2023


    Progress 07/01/20 to 06/30/21

    Outputs
    Target Audience:The target audience for this report are poultry producers as well as other commercial activities organized around poulty production. The major goal of this project is to produce safe and efficiacious live attenuated influenza virus vaccines against multiple avian influenza strains. In this particular application, the focus is on the prevention and controlof avian influenza viruses known as H5 and H9 subtypes which affect poultry production and food security inlarge areas of the world. The US is at great risk of "important" such devastating pathogens through many different routes including wild birds as exemplified by the 2015 outbreak of highly pathogenic avian influenza in the Midwest and the ongoing reports in 2022 of introductions of H5N1 in wild birds and outbreaks in poultry in multiple states.With minor modifications, we have produced the vaccines as proposed in the original application and are about to start safety and efficacy studies in chickens. Changes/Problems:We are on track and feel confident about the successful completion of the project. What opportunities for training and professional development has the project provided?The project has provided plenty of training opportunities in molecular biology and reverse genetics to graduate students (Lindsay Gay, Flavio Faccin, and Matias Cardenas) and two UGA Post-Bacculaureate Research Program(PREP) Scholars (Luis Rodriguez and Blake Vilchez). The project is largely run by Flavio Faccin. How have the results been disseminated to communities of interest?A manuscript is in preparation with data produced so far while we hope to produce data showing safety and efficacy of the vaccine constructs. What do you plan to do during the next reporting period to accomplish the goals?Complete cloning and rescue of recombinant vaccine virus candidates against the novel H5N1 virus and test stability. Coordinate with our Co-PI Dr. Darrell Kapzynski the safety and protection studies in chickens as per the original proposal (aim 2).

    Impacts
    What was accomplished under these goals? Despite the limitations related to the COVID-19 pandemic, aim 1 has been completed. We are focusing our efforts on H5 and H9 strains that continue to circulate in Egypt and present with a great opporunity to testing the vaccines for protection against wild type strains. With minor modifications all vaccine viruses as proposed in the application have been produced. Due to the multiple detection of H5N1 HPAIV in the US and accompanying outbreaks reported in poultry, we are producing a set of vaccine viruses against this novel strain. In the previous report, we reported some issues with the H9 HA construct. I am happy to report that such issue has been solved. All viruses grow to high titers and are stable. We are set to start aims 2 and 3 as proposed in the original application.

    Publications