Progress 07/15/19 to 07/14/23
Outputs
Target Audience:Target audience for this work include scientists and veterinary professionals working in the area of animal health, infectious disease, and vaccine design and developmentTarget audience for this work include scientists and veterinary professionals working in the area of animal health, infectious disease, and vaccine design and development. Changes/Problems:The COVIDsituation markedly affected progress of this work. What opportunities for training and professional development has the project provided?Project provided research experinces for three graduate students and one post-doctoral fellow. How have the results been disseminated to communities of interest?Presentations at professional meetinga and byresearsh publication. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Bovine papular stomatitis virus (BPSV) as a vaccine vector for cattle:Virus vectored vaccines are not available commercially for cattle even though compelling potential applications exist. Recombinant BPSV were constructed expressing either Bovine herpesvirus 1(BoHV-1)protective antigens glycoprotein gD (BPSVgD), or gD and gB (BPSVgD/gB). Immunization of BPSV serologically-positive calves with BPSVgDor BPSVgD/gBinduced BoHV-1 neutralization antibodies and provided protection for three of four animals following a high dose BoHV-1 challenge at day 70 pi.Results indicate BPSV suitability as a candidate virus vector for cattle vaccines. Notably and of likely consequence for potential future applications, the BPSV vector was effective in inducing immune responses in BPSV serologically-positive animals. This result is consistent with the known high reinfection potential of BPSV and its high prevalence and transmissibility in bovine populations; thus, prior BPSV infection does not pose a significant obstacle for a BPSV-based vector.BPSV vectors may offer unprecedented utility; an ability to establish persistent/latent infections, high host reinfection potential and highly efficient transmission characteristics may translate into "self-boosting vaccines" performing at both the individual and population level. A "self-boosting vaccine" would enhance the magnitude and duration of host immune responses to target antigens; this would be of particular advantage where longer duration immunity is desirable, such as in range production systems and dairy animals. Perhaps more importantly, these vectors may have potential as "perpetual vaccines" where a single vaccine dose will control target diseases within an animal population in perpetuity via "transmission immunization", thus delivering the benefits of vaccination at almost no cost. This strategy may reduce or remove the need for repeated "booster" vaccinations. Bovine herpesvirus 6 (BHV-6) vectoras a vaccine vector for cattle:A BHV-6 vector expressing BHV-1 gB/gD. Methods to work with highly cell-associated BHV-6 in cell culture were developed. Homologous recombination methods to engineer the BHV-6 genome using transfection/infection were developed and used successfully to construct and express trans-genes (ie.GFP). Unfortunately, efforts to construct a stable BHV-6 vector expressing BHV-1 gB/gD were unsuccessful.Recombinants were obtained initially, but were unstable on passage. Efforts to construct stable recombinant BHV-6 BAC clones expressing BHV-1 gB/gD also were unsuccessful. Incompatibility of inserting BHV-1 sequences into BHV-6 genome may be responsible. Further work will be required to resolve this issue
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Khatiwada, S., Chaulagain, S., Delhon, G. and Rock, D.L. 2019. Novel Viral Vaccine Vectors for Cattle. Conference for Research Workers in Animal Disease (CRWAD) P181; p.250.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2023
Citation:
Gustavo Delhon, Sushil Khatiwada, David Doub, Seth Harris, Sabal Chaulagain, Mostafa El-Gaffary, Daniel L. Rock. 2024. Bovine papular stomatitis virus as a vaccine vector for cattle. Conference for Research Workers in Animal Disease (CRWAD). To be presented January 2024.
- Type:
Journal Articles
Status:
Awaiting Publication
Year Published:
2023
Citation:
Gustavo Delhon; Sushil Khatiwada; David Doub; Seth Harris; Sabal Chaulagain; Mostafa El-Gaffary; Daniel L. Rock. Bovine Papular Stomatitis Virus as a Vaccine Vector for Cattle. Journal of General Virology (JGV-D-23-00190R) Accepted for publication October 19. 2023.
DOI 10.1099/jgv.0.001914
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Progress 07/15/21 to 07/14/22
Outputs
Target Audience:Target audience for this work include scientists and veterinary professionals working in the area of animal health, infectious disease, and vaccine design and development. Changes/Problems:As the COVID situation has markedly improved, we expect accelerated progress focused on careful evaluation of BPSV and BHV-6 vectors using vaccination/challenge experiments in calves. What opportunities for training and professional development has the project provided?Research conducted provided research training for one post-doctoral fellow and one PhD student. How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?Additional vaccination/challenge experiments with BPSVtgdD/tgdBare planned to further evaluate protective efficacy and evaluate persistence and transmissibility of the BPSVtgdD/tgdBvaccine vector in cattle. Efforts are continuing to construct stable recombinant BHV-6 BAC clones expressing BHV-1 gB/gD. Single gene constructs will be evaluated in an effort to improve BHV-6 vector stability.
Impacts
What was accomplished under these goals?
Objective: 1) Construction of GHV and PPV vectors expressing known protective antigens of BHV-1, glycoprotein B and D - Work continues on constructing stable Bovine herpesvirus 6 (BHV-6) BAC clones expressing BHV-1 gB/gD at high levels. Objective 2) Evaluation of protection afforded by vaccination with BPSVgB/gD1 using challenge infection in cattle. Promising results were obtained when the BPSVgB/gD1 vector was evaluated in calves. Animals were immunized with recombinant BPSV, C5 strain, expressing BHV-1 glycoproteins D and B lacking transmembrane domains (BPSVtgdD/tgdB) via intranasal route by nebulization (7.5 x 106TCID50) and IM (107TCID50) and boosted on days 21 and 49 post-priming. Immunized cattle seroconverted by day 28 post priming with serum neutralizing (SN) titers of 1:8 to 1:16, and titers of 1:16 to 1:128 were observed by day 56 post-vaccination.Antibodies directed against both gB and gD were detected in sera of vaccinated animals by western blot. No clinical signs associated with vector infection nor integumentary lesions associated with inoculation sites were observed, indicating that BPSVtgdD/tgdBis completely attenuated under vaccination conditions used. Preliminary data indicate that BPSVtgdD/tgdBvaccination resulted in protection (lack of clinical symptoms) of approximately 75% of animals following intranasal challenge withvirulent BHV-1 (Cooper strain) at 60 days post-vaccination.
Publications
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Progress 07/15/20 to 07/14/21
Outputs
Target Audience: Target audiences for this work include scientists and veterinary professionals working in the areas of animal health, infectious disease, and vaccine design and development. Changes/Problems:The COVID 19 situation markedly affected the pace of our laboratory/animal research during the past year.As the COVID situation has markedly improved,we anticipate accelerated progress in the coming year focused on carefully evaluating BPSV and BHV-6 vectors using vaccination/challenge experiments in calves. What opportunities for training and professional development has the project provided? Research conducted provided research training for one post-doctoral fellow and one PhD student. How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals? The availability of BPSVgB/gD1 will permit evaluation of its vaccine vector potential (attenuation, persistence, immunogenicity and transmissibility) in cattle. Additionally, efforts will be directed at constructing the (BHV-6) vector expressing BHV-1 gB/gD.
Impacts
What was accomplished under these goals?
Objective: 1) Construction of GHV and PPV vectors expressing known protective antigens of BHV-1, glycoproteins B and D - Efforts are ongoing to construct a Bovine herpesvirus 6 (BHV-6) vector expressing BHV-1 gB/gD. Issues related to recombinant virus instability and weak expression of clonedBHV-1, glycoproteins B and D geneswere encountered. Additional sites for foreign geneinsertionare being investigated to increaserecombinant virusstability. Also, a variety of BHV-6 early and late gene promoters are being screened to optimize high level BHV-1 gene expression in the context of the BHV-6 infected cell.Efforts to complete a BHV-6 BAC clone are ongoing.Objective 2)Evaluation of protection afforded by vaccination with BPSVgB/gD1and BHV-6gB/gD1using BHV-1 challenge infection in cattle.A bovine papular stomatitis virus (BPSV) vector expressing BHV-1 glycoproteins gB and gD of bovine herpesvirus 1 (BHV-1(BPSVgB/gD1) is currently being evaluated using vaccination/challenge experiments incalves to access protective immunity to BHV-1.
Publications
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Progress 07/15/19 to 07/14/20
Outputs
Target Audience:Target audiences for this work include scientists and veterinary professionals working in the areas of animal health, infectious disease, and vaccine design and development. Changes/Problems:The COVID 19 situation markedly affected the pace of our laboratory research as well as the rearing of the gnotobiotic animals necessary for vaccine evaluation. As the COVID situation is likely to improve by Spring/Summer 2021 we anticipate accelerated progress in the coming year. What opportunities for training and professional development has the project provided?Research conducted provided research training for one post-doctoral fellow and one PhD student?. How have the results been disseminated to communities of interest?Work was reported at the 2019 Conference of Research Workers in Anial Disease ( P181, p.250 of abstrack book).? What do you plan to do during the next reporting period to accomplish the goals?The availability of BPSVgB/gD1 will permit evaluation of its vaccine vector potential (attenuation, persistence, immunogenicity and transmissibility) in cattle. Additionally, efforts will be directed at constructing the (BHV-6) vector expressing BHV-1 gB/gD.
Impacts
What was accomplished under these goals?
Objective: 1) Construction of GHV and PPV vectors expressing known protective antigens of BHV-1, glycoproteins B and D - A bovine papular stomatitis virus (BPSV) vector expressing BHV-1 gD1(BPSV/gD1) or glycoproteins gB and gD of bovine herpesvirus 1 (BHV-1(BPSVgB/gD1) were constructed and evaluated for stable expression of BHV-1 gB/gD using immunofluorescence, immunoprecipitation and western blot. Both vectors exhibited high level expression in cell cultures. Efforts were initiated to construct a Bovine herpesvirus 6 (BHV-6) vector expressing BHV-1 gB/gD. Traditionl and a BAC-based strategy are currently being employed.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Khatiwada, S., Chaulagain, S., Delhon, G. and Rock, D.L. 2019. Novel Viral Vaccine Vectors for Cattle. Conference for Research Workers in Animal Disease (CRWAD) P181; p.250.
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