Progress 03/15/19 to 03/14/24
Outputs
Target Audience:
Nothing Reported
Changes/Problems:Because of the Covi-19 pandemic planned research was interrupted requiring extension of the grant period to obtain additional time to complete ongoing studies. What opportunities for training and professional development has the project provided?The project has provided opportunities for 3 post doctoral associates to gain training for careers in veterinary immunology. How have the results been disseminated to communities of interest?The results are being dissemenated through publication of data and participation in national meetings. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Studies are still ongoing. As indicatedin the title of this project, emphasis was placed on developing and using additional mAb reagent with emphasis on use in cattle. Additional mAb reagents have been deveoped for use in the study of the immune response to pathogens using M. a. paratuberculosis (the causative agent of Johne's disease in cattle and Crohn's disease in humans). These include regulating the the immune response: CD27, CD95, CD152,CD154, CD178, and CD179. It also inludes mAbs tokey cytokines: IFN-gamma, TNF-alpha, IL-6, IL-8, IL-10, IL-17, and IL-22. All the mAbs are being made available through the university monoclonal antibody center.
Publications
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Progress 03/15/22 to 03/14/23
Outputs
Target Audience:Investigators interested and/or conducting research on the immune response to pathogens,parasites and development of vaccines. Changes/Problems:As mentioned in our second year report, our overall productivity was severly impacted by the covid 19 pandemic. Because of the pandemic there was a university wide requirement to reduce or stop all research activity. In addition for the university order, we encountered an additional unexpected problem. We lost over a year do to these events. We requested and received an extension of the grant period to continue our studies. Because of the impact of covid 19, we still need additional time to continue the studies proposed in this grant. Funds have been conserved in the grant for this purpose. As noted in the plans for the coming year, the associates from Egypt are back so it will be possible to complete the proposed studies with extension of grant period for one more year. What opportunities for training and professional development has the project provided?The research program has provided opportunity for research associates to increase background knowledge on the immunology of infectious diseases with a focus on mycobacterial pathogens. The program has also provided opportunity to help improve writing skills through writing up results from ongoing studies. How have the results been disseminated to communities of interest?Results from ongoing studies have been published in scientific journals. What do you plan to do during the next reporting period to accomplish the goals?The plans for the coming year are several fold: Continue validating specificity of mAbs developed before and during the grant period. This includes CD152, CD154, IL-12R, and IL-23R. Continue efforts to develop mAbs against CD95, CD178, CD173, CD174, GARP. Continue efforts to use mass spectrometry to determenethe specificity of a set of mAbs developed against molecules expressed on activated bovine lymphocytes: Temporary names ACT2A, ACT9A, neutrophils. Validate mAbs developed against bovine leukocytes for use with yak (closely related cattle), bison, deer, and elk.
Impacts
What was accomplished under these goals?
Studies were resumed on return of research my research associates from Egypt the beginning of 2022. The closing of the embassy in Egypt for issuing visas until the end of 2021, because of covid virus, delayed their return for a year. 1) The focus has been on the further documentation of the functional activity of some of the mAbs, developed during the grant period, in characterizing the immune response to mycobacterial pathogens (manuscript under review). The monoclonal antibodies were also used to document the immune response to a candidate virus vectored peptide for paratuberculosis (manuscript in preparation). 2) Further studies were conducted to document the functional activity of monoclonal antibodies against CD152 and CD154. For monoclonal antibody production, monclonal antibodies were produced against IL-12R and IL-23R the receptors for IL-12 and IL-23. Studies are planned for documenting specificity and function. 3) The universal platformdeveloped to study the primary and recallimmune responses Mycobacterial and other pathogens was described and published to make the technology available to the research community. 4) The monoclonal antibodies developed developed before and during the grant period were used to obtain further information showing the inability of mycobacterial pathogens to survive in absence of a single gene (rel) is attributable to clearance of bacteria from all tissues by development of CD8 cytotoxic T cells. (manuscript under review)
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2022
Citation:
Elnaggar MM, Abdellrazeq GS, Sacco RE, Harsla TR, Mucci ML, Fry LM, Hulubei V, Davis WC. 2022. Comparative analysis of the specificity of monoclonal antibodies developed against the bottlenose dolphin, Tursiops truncatus, TNF-alpha, IL1-beta, IL-6, IL-8, IL-10 with monoclonal antibodies made against ovine IFN-gamma bovine IL-17A and IL-1beta revealed they recognize epitopes conserved on dolphin and bovine orthologues. Vet Immunol Immunopathol 250:110456.Vet Immunol Immunopathol 250:110456.
- Type:
Journal Articles
Status:
Published
Year Published:
2022
Citation:
Davis WC, Mahmoud AH, Abdellrazeq GS, Elnaggar MM, Dahl JL, Hulubei V, Fry LM. 2022. Ex vivo Platforms to Study the Primary and Recall Immune Responses to Intracellular
Front Vet Sci 9:878347. Volume 9 - 2022 | https://doi.org/10.3389/fvets.2022.878347
Mycobacterial Pathogens and Peptide-Based Vaccines.
- Type:
Journal Articles
Status:
Under Review
Year Published:
2023
Citation:
Asmaa H. Mahmoud, Gaber S. Abdellrazeq, Lindsay M. Fry, David A. Schneider, Kun Taek Park, Sarah Attreedf, Waithaka Mwangig, Leeanna Burtong, Neha Sangewarg, Cleverson deSouza, Victoria Hulubei, and William C. Davis. Identification of rel, a gene regulating the stringent response, as Achilles heel for mycobacterial pathogens. MDPI Vaccines (under review).
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Progress 03/15/21 to 03/14/22
Outputs
Target Audience:Stake holders livestock production. Research investigators involved in investigating immune response to pathogens and development of vaccines. Changes/Problems:As mentioned in our second year report, our overall productivity was severly impacted by the covid 19 pandemic. Because of the pandemic there was a university wide requirement to reduce or stop all research activity. In addition for the university order, we encountered an additional unexpected problem. The international J1 research scholar, G. Abdelrazeq, that we had trained to conduct the research outlined in the current project had to return to Egypt to update his visa status. The US embassy was closed anddirected to stop renewing and/or issuing visas for travel to the US. G. Abdelrazeq was left stranded in Egypt for the past year and a half. His opportunity to return to the US is now projected to be the end of Jan. 2022. A request is being made for an extension of the current grant to provide an opportunity to continue the objectives of grant. A request through the WSU Office of Research Support and Operations for a one year extension of grant 20186701528744 ORSO# 129153-006 has been approved. November 24, 2021 Awards Management Branch Office of Extramural Programs U.S. Department of Agriculture, NIFA 1400 Independence Ave SW Stop 2271 Washington DC 20250?2271 RE: USDA Agreement Number: 20186701528744 Good day, ORSO # 129153?006 Washington State University has approved a no?cost extension for twelve (12) months for the above? referenced grant. The termination date has been extended from 3/14/2022 to 3/14/2023 The extension is necessary to complete the project. This does not change the scope of the grant. The post docs were stranded in Egypt trying to get their visas renewed. Asmaa succeeded in getting an emergency appointment at the embassy and got her visa renewed. Gaber's renewal was delayed and now he plans to return in another 50 days so that he is eligible for a J1 visa again. Asmaa was able to return a couple of weeks ago to help get the research in motion again. This notification is being submitted in compliance with Article 14 of the Research Terms and Conditions, dated October 2016. Sincerely, Dan Nordquist Dan Nordquist Associate Vice President Authorized Signatory What opportunities for training and professional development has the project provided?Our opportunities to continue our training program for visiting scholars and graduate students were interrupted by the covid 19 pandemic and requirement by the university to reduce or stop research activity. How have the results been disseminated to communities of interest?Abdellrazeq, G.S., Fry, L.M., Elnaggar, M.M., Bannantine, J.P., Schneider, D.A., Chamberlin, W.M., Mahmoud, A.H.A., Park, K.T., Hulubei, V., Davis, W.C., 2020a. Simultaneous cognate epitope recognition by bovine CD4 and CD8 T cells is essential for primary expansion of antigen-specific cytotoxic T-cells following ex vivo stimulation with a candidate Mycobacterium avium subsp. paratuberculosis peptide vaccine. Vaccine 38, 2016-2025. Abdellrazeq, G.S., Mahmoud, A.H., Park, K.T., Fry, L.M., Elnaggar, M.M., Schneider, D.A., Hulubei, V., Davis, W.C., 2020b. relA is Achilles' heel for mycobacterial pathogens as demonstrated with deletion mutants in Mycobacterium avium subsp. paratuberculosis and mycobacterium bovis bacillus Calmette-Guerin (BCG). Tuberculosis (Edinb) 120, 101904. Davis, W.C., Abdellrazeq, G.S., Mahmoud, A.H., Park, K.T., Elnaggar, M.M., Donofrio, G., Hulubei, V., Fry, L.M., 2021. Advances in Understanding of the Immune Response to Mycobacterial Pathogens and Vaccines through Use of Cattle and Mycobacterium avium subsp. paratuberculosis as a Prototypic Mycobacterial Pathogen. Vaccines (Basel) 9. What do you plan to do during the next reporting period to accomplish the goals?Continue and complete characterization of mAbs developed against CD27, CD152, CD154, TNFa, IL-6, IL-17, IL-10, IL-17, IL-22, granulysin. Complete cloning and validation of mAbs against IL-12R and IL23R. Develop mAbs against CD95, CD178, CD274,and CD179.
Impacts
What was accomplished under these goals?
Futher studies were conducted to validate mAbs developed during the first and second year of grant. CD27, CD152, CD154, TNFa, IL-6, IL-17, IL-10, IL-17, IL-22, granulysin. Efforts were initiated to develop monoclonal antibodies to IL-12R and IL-23R. Studies are in progress to identify and clone hybridoma cell lines producing these monoclonal antibodies.Because of the covid 19 pandemic progress has beenslowed in part becase of a university wide requirement to reduce or stop ongoing research.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Davis, W.C., Abdellrazeq, G.S., Mahmoud, A.H., Park, K.T., Elnaggar, M.M., Donofrio, G., Hulubei, V., Fry, L.M., 2021. Advances in Understanding of the Immune Response to Mycobacterial Pathogens and Vaccines through Use of Cattle and Mycobacterium avium subsp. paratuberculosis as a Prototypic Mycobacterial
Pathogen. Vaccines (Basel) 9
William C. Davis, Gaber S. Abdellrazeq, Asmaa H. Mahmoud, Kun-Taek Park, Mahmoud M. Elnaggar, Victoria Hulubei, Lindsay M. Fry. Demise of a myth: The macrophage is not a safe-haven for mycobacterial pathogens. CRWAD 2021
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Progress 03/15/20 to 03/14/21
Outputs
Target Audience:
Nothing Reported
Changes/Problems:Progress in development of mAbs was interrupted by the covid-19 pandemic limiting personnel activities in the laboratories and the difficulty of my Egyptian post doc in getting his visa renewed to return to the US to continue participating in development and characterization of mAbs. Full continuation of laboratory activities on mAb development will be difficult until the pandemic is brought under control. What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest?Manuscripts are in progress describing the development and validation of the mAbs developed in the first and current year. What do you plan to do during the next reporting period to accomplish the goals?We expect to increase productivity on the development of mAbs to the molecules listed in the project proposal for development. Genes encoding some of the molecules have been expressed and are ready to be used to express the molecules for mAb production.
Impacts
What was accomplished under these goals?
Progress in developing monoclonal antibodies for use in the study of the immune response to pathogens and vaccine development were interrupted by the covid-19 pandemic and the closing of the US embassy in Egypt. My Egyptian post. doc has been stranded in Egypt trying to get his visa renewed. We were able to develop one mAb (CD154) before he returned to Egypt to renew his visa. In preparation for his return, mice were immunized to develop mAbs to IL-12R, IL-21R, and IL-23R. The spleens from the immunized mice were collected and cryopreserved for later use in the development of mAbs to IL-12R, IL-21R, and IL-23R. We expect to resume mAb development 2-1-2021. Manuscripts describing the production of mAbs to IL-6, IL-8, IL-10, IL-22, TNF-a are in progress.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2020
Citation:
Simultaneous cognate epitope recognition by bovine CD4 and CD8 T cells is essential for primary expansion of antigen-specific cytotoxic T-cells following ex vivo stimulation with a candidate Mycobacterium avium subsp. paratuberculosis peptide vaccine CRWAD 2020
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Progress 03/15/19 to 03/14/20
Outputs
Target Audience:The objectiveproposed to initiate the current project on development of monoclonal antibodies (mAb) to bovine leukocyte differentiation molecules (LDM) was to express LDM and cytokines encoded by CD83, CD95, CD152, CD154, CD178, CD192, CD197, CD273, CD274, CD279, CD223, CD282, CD284, CD366, IL-22, IL-12R, IL-23R. CD95, CD152, CD154, CD178, CD279, CD282, IL-22, IL12R, and IL-23R were expressed for mAb production. mAbs were successfully developed against CD152 and IL-22. In addition, mAbs were successfully developed against molecules not on the list proposed to initiate the project: CD27, TNF-a, IL-6, IL-8 andIL-10. Documentation of specificity of the mAbs is in progress. Manuscripts will be prepared for publication as soon as documentation studies are complete. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Training in mAb development, flow cytometry, and analysis of the immune response was provided to 3 post doctoral associates. How have the results been disseminated to communities of interest?As reported above, methodology and use of mAbs developed for research in cattle have been publised. What do you plan to do during the next reporting period to accomplish the goals?As mentioned above, genes encodingmolecutes of interest for developing mAbs have been expressed. These molecules will be used to immunize mice for mAb development. Further training of post doctoral associates will be continued focused on gaining expertese in analysis of the immune response to pathogens.
Impacts
What was accomplished under these goals?
Under objective 1, we proposed to initiate the current project on development of monoclonal antibodies (mAb) to bovine leukocyte differentiation molecules (LDM) starting with expression ofLDM and cytokines encoded by CD83, CD95, CD152, CD154, CD178, CD192, CD197, CD273, CD274, CD279, CD223, CD282, CD284, CD366, IL-22, IL-12R, IL-23R. CD95, CD152, CD154, CD178, CD279, CD282, IL-22, IL12R, and IL-23R were expressed for mAb production. mAbs were successfully developed against CD152 and IL-22. In addition, mAbs were successfully developed against molecules not on the list proposed to initiate the project: CD27, TNF-a, IL-6, IL-8 andIL-10. Documentation of specificity of the mAbs is in progress. Manuscripts will be prepared for publication as soon as documentation studies are completed. Under objectives 2 and 3: We have developed assays to study the antigen processing by antigen presenting cells and analyisis of the functional activity of CD4 helper and CD8 effector T cells that develop in response to antigen presentation by antigen presenting cells. the methodologies have been described in our recent publications. Abdellrazeq GS, Fry LM, Elnaggar MM, Bannantine JP, Schneider DA, Chamberlin WM, Mahmoud AHA, Park KT, Hulubei V, Davis WC. Simultaneous cognate epitope recognition by bovine CD4 and CD8 T cells is essential for primary expansion of antigen-specific cytotoxic T-cells following ex vivo stimulation with a candidate Mycobacterium avium subsp. paratuberculosis peptide vaccine. Vaccine 2020 doi: 10.1016/j.vaccine.2019.12.052 Franceschi V, Mahmoud AH, Abdellrazeq GS, Tebaldi G, Macchi F, Russo L, Fry LM, Elnaggar MM, Bannantine JP, Park KT, Hulubei V, Cavirani S, Davis WC, Donofrio G. Capacity to Elicit Cytotoxic CD8 T Cell Activity Against Mycobacterium avium subsp. paratuberculosis Is Retained in a Vaccine Candidate 35 kDa Peptide Modified for Expression in Mammalian Cells. Front Immunol 2019;10:2859. doi: 10.3389/fimmu.2019.02859 Abdellrazeq GS, Elnaggar MM, Bannantine JP, Schneider DA, Souza CD, Hwang J, Mahmoud AHA, Hulubei V, Fry LM, Park KT, Davis WC. A peptide-based vaccine for Mycobacterium avium subspecies paratuberculosis. Vaccine 2019;37:2783-2790. doi: 10.1016/j.vaccine.2019.04.040
Publications
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