Progress 09/01/13 to 08/31/17
Outputs
Target Audience:During the life of this project we interacted with three categories of audiences: 1) Scientific academic audiences( scientists and graduate students ): we have consistently contributed presentations and received invitations to give scientific expositions at animal health annual forums, such as the Conference of Research Workers in Animal Diseases (CRWAD), the American Society of Virology annual meeting, the NIFA Animal Health Project Directors' annual meeting, the International Pig Veterinary Society bi-annual meeting, The NC229 CSRS annual meeting ( PRRSV and other emerging diseases of swine). In addition at least 6 peer review publications derived from this project have been produced, the number of citations resulting from these publications keeps increasing on a constant basis providing \us with good estimator of the positive impact of the results of the project. 2) Swine industry and swine practitioner organizations: such as National Pork Board(NPB), which has annually received and positivel evaluated submissionsderivedfrom preliminary results produced from this project. The NPB board of governors and other administrative authorities of NPB have contacted PD and co-PDs to serve as external experts in topics related to this project. 3) Biologics companies and intellectual property agencies related to biologics inventions: given the fact that an important patent licensed by Nutech Ventures (IP agency of the University of Nebraska)has beenpartly derived from this project, Nutech Ventures organized marketing and promotion of our invention throughout an important group of biological companies. The R and D units of these companies invited us to give presentations and/or exhibits on the novelties of our invention. A principal outcome of this IP management strategy by Nutech is signaled by the current signing of an exclusive license with one of those companies for severalyears to come. : Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Three doctoral programshave beensuccesfully completed under this project. On december 2017 these three graduate students will officially receive their PhD degrees in the general area of Virology - specific programs: IBMS integrative biomedical sciences (2) and School of Biological Sciences (1)- granted all of them by the University of Nebraska-Lincoln. These three new PhDs have signed as first author on 4 of the 6 peer reviewed articles produced by this project. One of the newly graduated PhDs has been already hired by a major international veterinary biologics company based in Michigan. A secodn graduating PhD is also exploring employment with a veterinary biologics company. The thirdnew PhD is already enrolled to continue in a viral pathogenesis ( animal virology) postdoctoral research program at the Nebraska Center for Virology. During the life of thisproject, and while working directly on the research herein reported, onepostdoctoralcandidateadvanced in his scientific career, becoming first a non-tenured research faculty , and later becoming a tenure track faculty member in one department (Animal Scince) at the University of Nebraska-Lincoln. How have the results been disseminated to communities of interest?1) Publications in peer reviewed journals: (6) 2) Presentations at annual meetings ( 2014 to 2017) of the North American PRRSV Symposium , Conference of Research Workers in Animal Diseases, American Society for Virology. 3) University of Nebraska internal graduate students seminars, posters competitions, and general public presentations and videos produced by Nutech Ventures (Intellectual Property Agency of the University of Nebraska-Lincoln) What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Derived from the experiments conducted under this project, three major concepts can be listed: 1) The synthesis of inter-genotype chimera( PRRSV 1 X PRRSV 2) that permits to study the hurmoral and and cellular immune reponse specific for the structural trimer of minor glycoproteins of PRRSV. This chimera permits presenting ,to the pig's immune system, the immunogenically important PRRSV trimer (GP2-3-4) in a natural form such that it may conserve important conformational epitopes within the context of a replicating vector. 2) The use of the above-mentioned chimera for a tolerization strategy that would permit developing potential broadly neutralizing monoclonal antibodies against PRRSV ( such concept is the basis of a proposal recently submitted to NIFA animal health program as the logical continuation of the project herein terminated/reported). 3) Confirmation that GP2-gp3 and Gp4 may be the central structural components PRRSV essentail for evoking broadly neutralizing antibodies. In summary the results of this project should have significant bearing on the development of new generation, subunit (non-replicating) vaccines against PRRSV
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2017
Citation:
Cross reactivity of immune responses to porcine reproductive and respiratory syndrome virusinfection.
Correas I, Osorio FA, Steffen D, Pattnaik AK, Vu HL.
Vaccine. 2017 Feb 1;35(5):782-788. doi: 10.1016/j.vaccine.2016.12.040. Epub 2017 Jan 3.
PMID: 28062126
- Type:
Journal Articles
Status:
Published
Year Published:
2017
Citation:
Relative contribution of porcine reproductive and respiratory syndrome virus open reading frames 2-4 to the induction of protective immunity.
Kimpston-Burkgren K, Correas I, Osorio FA, Steffen D, Pattnaik AK, Fang Y, Vu HLX.
Vaccine. 2017 Aug 3;35(34):4408-4413. doi: 10.1016/j.vaccine.2017.06.061. Epub 2017 Jul 6.
PMID: 28689650
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Progress 09/01/15 to 08/31/16
Outputs
Target Audience:During the third year of this project we have communicated with the following target audiences: 1) Academic : we continued active laboratory hands-on training for three graduate students and provided project plans for one senior research associate and one international resaerch scientist 2) Industry: we have provided support toi one company exploring a patent developed by two of us that is being explored by an international vaccine manufacturing company 3) Scientific professional: we have made two presentations at a major national meeting in the area of animal agriculture(animal health) and gave key note presentations based directly on the results of this project, at 2 international forums: International Pig Veterinary Society ( Dublin, Ireland) , and FAVA ( Federation of Asian Veterinary Medical Associations) Ho Chi Minh City Vietnam. Changes/Problems:None for this year What opportunities for training and professional development has the project provided?The objectives of this proposal are providing thesis research themes for 3 doctoral candidates and one independent project for a senior research associate How have the results been disseminated to communities of interest?2 presentations by graduate students at the Conference of Research Workers in Animal Disease and the 2016 North American PRRS International Symposium: K. Kimpston-Burkgren1,3*, H. Vu2,3, I. Correas2,3, D. Steffen2, A. Pattnaik2,3, Y. Fang4, F. Osorio2,3. Contribution of PRRSV minor glycoproteins to a protective immune response in swine School of Biological Sciences, University of Nebraska-Lincoln, Lincoln, NE. 2School of Veterinary Medicine and Biomedical Sciences, University of Nebraska-Lincoln, Lincoln, NE. 3Nebraska Center for Virology, Lincoln, NE. 4Department of Diagnostic Medicine/Pathology, Kansas State University, Manhattan, KS. I Correas, Osorio FA, Pattanik A, Vu hiep Cross-reactivity of immune responses against Porcine Reproductive and Respiratory Syndrome Virus School of Veterinary Medicine and Biomedical Sciences, University of Nebraska-Lincoln, Lincoln, NE. 3Nebraska Center for Virology, Lincoln, NE. What do you plan to do during the next reporting period to accomplish the goals?We anticipate that this 4th year extended research will lead to succesful close the 3 PhD student projects. We anticipate that this research will pemit conclusion and publication of at least 3 new refereed articles during the period 2017-2018.
Impacts
What was accomplished under these goals?
In all likelihood, the major impact of the research results obtained during this third year of the proyect derives from our implementation of a new strategy that would permit using a heterologous PRRSV genotype strain as its own replicating vector to advance knowledge on the fine molecular and epitope structure of PRRSV that have beenpursued along the 3 objectives of this project. A particularly attractive feature of our model isthat of using one genotype of PRRSV ( type 1 European genotype) as chimeric vehicle or vector of specificproteins or antigens of the other PRRS genotype (PRRS type 2 northAmerican genotype).The reports of our resultsusing such chimeras to providean accuratequantitation of the level of protection conferred by each of these individual gentotype 2 proteins (GP2, GP3 and GP4) have been competitively selected for oral presentations (and awarded travel funds to each of the 2 presenting students) to begiven at the upcoming North-American Symposium on PRRS and Other Swine Emerging Viruses. I addition, this technique of using chimeras between genotype I and genotype 2 of PRRSVrepresents a particularly interestingvaccinology modelfor those countries or regions that are currently pursuing the possible use of vaccines or immunogens that are based on inter-genotype cross protection, like is the case ofcertain Far-Eastern countries.Inareas like Korea or Northern China, swine farms are significantly affected by PRRSV strains belongingto both genotypes. Therefore that explains our current results havingbeen invited for main keynote presentation at the 18th Annual meeting of FAVA ( Federation of Assian Veterinary Med Associations, a regional organization involving 28th Assian countries) .Our results, which generated a significant number of interest and questions at such meeting, served toshow to this audience that the intergenotype cross protection appraoch is very limited in efficacy. That was probably the main take-home message obtained from these results that was very useful for such regional audience. A summary of the research steps pursued during 2015-2016 (third year of the project) for the objectives 1 and 2 follows: Objective 1: to explore alternate vectors different than the ones we originally planned when submitting this proposal, and to explore a more effective prime-boosting immunization with individual glycoproteins(GPs) of PRRSV by giving priority to the simultaneous expression of all 3 minor GPS in the same cells, rather than separatedly expressing the 3 individual proteins. To ensure the most effective replication and expression of the gP type 2 PRRSV in live replicating vector we have developed chimeric PRRSV viruses that consist of PRRSV type 1 vectors (Euro PRRS) expressing GPs of type 2. Objective 2: To map the structural basis of the important cross-protective epitopes( B and T) in different PRRSV antigens we are now expressing all the possible genes of PRRSV ( structural and non-structural total (n= 16) in recombinant bacterial expression systems or in multipeptide synthgetic preparations to be used to cross stimulate T cells from PRRSV-immunized pigs. This technique permitetd us to estimate the relationship of PRRSV genetic diversity among multiple type strains using vrius neutralization and gamma IFn producing cell assays.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Identification of viral genes associated with the interferon-inducing phenotype of a synthetic porcine reproductive and respiratory syndrome virus strain.
Sun H, Pattnaik AK, Osorio FA, Vu HL.
Virology. 2016 Oct 10;499:313-321. doi: 10.1016/j.virol.2016.09.018. [Epub ahead of print]
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Complete Genome Sequence of Highly Virulent Porcine Reproductive and Respiratory Syndrome Virus Variants That Recently Emerged in the United States.
Workman AM, Smith TP, Osorio FA, Vu HL.
Genome Announc. 2016 Aug 4;4(4). pii: e00772-16. doi: 10.1128/genomeA.00772-16.
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Strategies to broaden the cross-protective efficacy of vaccines against porcine reproductive and respiratory syndrome virus.
Vu HL, Pattnaik AK, Osorio FA.
Vet Microbiol. 2016 Sep 21. pii: S0378-1135(16)30365-0. doi: 10.1016/j.vetmic.2016.09.014. [Epub ahead of print]
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Progress 09/01/14 to 08/31/15
Outputs
Target Audience:During the second year of this project we have communicated with the following target audiences: 1) Academic : we continued active laboratory hands-on training for three graduate students and provided project plans for one senior research associate 2) Industry: we have produced preliminary data that permitted to obtain one successful award for a collaborative proposal with South Dakota State University. Such award was granted by a commodity group (National Pork Board) 3) Scientific professional: we have made two presentations at a major national meeting in the area of animal agriculture (animal health) and gave presentations at 3 international forums: at Seoul National University, and Konkuk University (both in Korea) and in Kyoto (Japan). Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Training activities: The objectives of this proposal are providing thesis research themes for 3 doctoral candidates and one independent project for a senior research associate How have the results been disseminated to communities of interest? We have been disseminating results by giving invited oral presentation to biologics companies (veterinary biologics) and by giving oral presentation to the scientific /veterinary researchers community (CRWAD, Chicago IL, Foreign Universities (korea Japan) What do you plan to do during the next reporting period to accomplish the goals?We plan to continue ongoing work in objectives 1 2 and 3 simultaneously
Impacts
What was accomplished under these goals?
The main impact of this project is based in that our research goals include a systematic study and characterization of the mechanisms involved in cross protection by PRRS viruses and by PRRS vaccines. The cross protection against PRRSV, which is the ability to simultaneously induce immunity against multiple diverse strains of PRRSV that currently act in the field ( ie. goal: "universal vaccine") is a principal priority identified by the National Pork Board (NPB). NPB is the major commodity group representing the pork industry. In the second year of this project, perhaps the major accomplishment that deserves to be cited along this project is undoubtedly consequence of having continued the changes and optimizations proposed last year for objective 1 , which were directed at optimizing different strategies of PRRSV glycoprotein immunization to broaden protective immunity against PRRSV. During 2015, our current research along this line has provided additional new data that has let our lab to competitively attract additional competitive to the university of Nebraska in 2015. Such new funds were awarded by swine industry, sp,. the National Pork Board, and are directly targeted towards a ideal concrete practical outcome for industry: a new vaccine, the award is entitled : Project ID 30581: Development of a live-attenuated PRRSV strain capable of eliciting a broad spectrum of heterologous protection, Hiep Vu (PI) and F A Osorio (co-PI) , NPB October 1 2015/September 30, 2016, amount :$ 100,000 A summary of the research steps pursued during 2014-2015 (second year of the project) for the objectives 1 and 2 follows: Objective 1: Last year we have we have found that is was necessary to explore alternate vectors different than the ones we originally planned when submitting this proposa, and to explore a more effective prime-boosting immunization with individual glycoproteins(GPs) of PRRSV by giving priority to the simultaneous expression of all 3 minor GPS in the same cells, rather than separatedly expressing the 3 individual proteins. To ensure the most effective replication and expression of the gP type 2 PRRSV in live replicating vector we have developed chimeric PRRSV viruses that consist of PRRSV type 1 vectors (Euro PRRS) expressing GPs of type 2. The evaluation of these live constructs is currently ongoing. Objective 2: To map the structural basis of the important cross-protective epitopes( B and T) in different PRRSV antigens we are now expressing all the possible genes of PRRSV ( structural and non-structural total (n= 16) in recombinant bacterial expression systems with the intent that the expressed products to be used to stimulate T cells from PRRSV-immunized animals.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2015
Citation:
Loving CL, Osorio FA, Murtaugh MP, Zuckermann FA.(2015) Innate and adaptive immunity against Porcine Reproductive and Respiratory Syndrome Virus.Vet Immunol Immunopathol : S0165-2427(15)00146-4. doi: 10.1016/j.vetimm.2015.07.003. [Epub]
- Type:
Journal Articles
Status:
Under Review
Year Published:
2015
Citation:
Vu,HLX Ma,F, Laegreid,WW, Pattnaik,AK, Steffen,D,Doster,A, and Osorio FA (2015) A synthetic porcine reproductive and respiratory syndrome virus strain confers unprecedented levels of heterologous protection JVI01657-15
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Progress 09/01/13 to 08/31/14
Outputs
Target Audience: During the first year of the project we have reached the following target audiences: 1) Academic: we have started active laboratory training for three graduate students and project plans for one senior research associate. 2) Industry: we have provided preliminary data that permitted to obtain two succesful awards from proposals submitted in 2014 to a commodity group (National Pork Board) 3) Industry: we have presented our initial results to a major biologics company 4) Scientific/Professional: we have made a presentation at a major national meeting in the area of animal agriculture ( animal health) Changes/Problems: In objective 1, we found that is was necessary to explore alternate vectors different than the ones we originally planned when submitting this proposal. The rationale for making such change is to achieve a better, more effective prime-boosting immunization with individual glycoproteins(GPs) of PRRSV. Therefore, we now give priority to the simultaneous expression of all 3 minor GPS in the same cells, rather than separatedly expressing the 3 individual proteins. This change does not alter the the ultimate goals of this proposal. What opportunities for training and professional development has the project provided? Training activities: The objectives of this proposal are providing thesis research themes for 3 doctoral candidates and one independent project for a senior research associate How have the results been disseminated to communities of interest? We started disseminating results by giving invited oral presentation to biologics companies ( Merial, veterinary biologics) and by giving oral presentation to the scientific /veterinary researchers community (CRWAD, Chicago IL) What do you plan to do during the next reporting period to accomplish the goals? We plan to continue ongoing work in objectives 1 2 and 3 simultaneously
Impacts
What was accomplished under these goals?
The impact of this project is represented by the fact of this being one of the few competitive research USDA-funded projects that systematically study and characterize the mechanisms involved in cross protection by PRRS viruses and by PRRS vaccines. The cross protection against PRRSV, which is the ability to simultaneously induce immunity against multiple diverse strains of PRRSV that currently act in the field ( ie. goal: "universal vaccine") is a principal priority identified by the national Pork Board (NPB). NPB is the major commodity group representing the pork industry. In the first year of this project we have started its 3 objectives simultaneous and independently, with a realistic degree of progress in each of them (see below). However, the major achievement accomplished during this first year is undoubtedly consequence of the changes and optimization that our lab , as part of objective 1 ( see below) has made to our classical paradigm of animal trials to assess cross protective immunity in pigs. The changes and modifications made at the start of objective 1, has permitted our lab to attract additional competitive awards to the university of Nebraska in 2014. Those funds were awarded by swine industry, sp,. the national pork Board, and these are: 1) Project ID 28693: Determine the Mechanisms of Cross-Protection against Infection with a Divergent Porcine Reproductive and Respiratory Virus Strain RFP Title: NPB Spring Call 2014 PI: Hiep Vu, & Osorio Co-Pi, $ 100,000 2) Project ID 28696: Evaluation of Conserved B- and T-Cell Epitopes as Inducers of Protective Immunity against Porcine Reproductive and Respiratory Syndrome Virus RFP Title: NPB Spring Call 2014 Sub-contract to South Dakota State University UNL Faculty: Hiep Vu & Osorio FA $26,945 A summary of the reaearch steps followed in each of the 3 objectives for the first year of the project follow: Objective 1: Using heterologous prime-booster immunizations based on different vectors we started testing the relative ability of each of these proteins to induce protective immunity. We found that is was necessary to explore alternate vectors different than the ones we originally planned when submitting this proposal.The rationale for making such change is to achieve a better, more effective prime-boosting immunization with individual glycoproteins(GPs) of PRRSV.Therefore, we now give priority to the simultaneous expression of all 3 minor GPS in the same cells, rather than separatedly expressing the 3 individual proteins. This change does not alter the the ultimate goals of this proposal. Objective 2: We focused on the interaction of PRRSV GP4 protein with the CD163 receptor through a mapping strategy that uses deletion and amino acid substitutions. We have so far good expression of the plasmids expressing GP4 and CD 163, which provides us with a bona fide positive control for future experiments in year 2 of the project. Objective 3: Started basic B cell cloning from swine, focusing on the expression of heavy and lIght chains of swine immunoglobulins.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2013
Citation:
Identification of a potentially cross protective porcine reproductive and respiratory syndrome virus strain B. Kwon*, H.L.X. Vu, W.W. Laegreid, T.K. Anderson, T.L. Goldberg, J. Christopher-Hennings, E.A. Nelson, F. Cerutti, K.M. Lager, A. Doster, A.K. Pattnaik, F.A. Osorio. Abstract No. 148, page 84, Proceedings 94th Annula Meetg CRWAD, December 9, 2013 Chicago Illinois
- Type:
Other
Status:
Other
Year Published:
2014
Citation:
PRRSV immunology & vaccines Osorio F Oral invited presentation PRRSV R & D Global Forum, Merial Inc., Atlanta, GA Sept 11 2014 (ppt circulated)
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