Progress 03/01/11 to 02/28/15
Outputs
Target Audience: Target audiences included veterinarians, scientists, biologics companies and agriculture or veterinary faculty. This was accomplishedby thevirtual PD workshop for animal health NIFA grantees, and the NC1192 meeting for bovine respiratory disease research. Individual consults also occured. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? Joe Agnes, PhD and Changyou Lin, DVM, PhD are two post-doctoral scientists trained in the Corbeil Lab under this grant. One pre Vet student, Ann Sung, also received training in the Corbeil lab. Heather McEligot and Nicole Behrens received training as graduate students in the Gershwin Lab. Matt Shao, a project scientist in the Gershwin lab, was involved in professional development which resulted in the submission of a first author manuscript. How have the results been disseminated to communities of interest? The results have been reported at professional meetings and communicated to biologics companies and practicing veterinary pathologists. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Objective 1. The hypothesis that BRSV increases sensitivity of BAT2 cells to H. somni cytotoxicity was tested in tissue culture. Cells treated with BRSV and H. somni shed virulence factors in concentrated culture supernatant (CCS) retracted more than cells treated with either agent alone. Paracellular migration of H. somni was also enhanced in dual treated cells. Objective 2. Microarray analysis of BAT2 cells treated with BRSV and CCS or either treatment alone, showed that several cytokines as well as MMPs were expressed at higher levels after dual treatment than with either agent alone. Protein ELISA assays confirmed that cytokine and MMP protein levels were both enhanced. Functional studies followed. When H. somni CCS alone was used, antiviral protein expression was up-regulated. Objective 3. Studies of immunomodulation in calves have been done and immunologic assays essentially completed. Data is currently being analysed.
Publications
- Type:
Journal Articles
Status:
Submitted
Year Published:
2015
Citation:
D. OToole, R. Hunter, T. Allen, B. Zekarias, J. Lehmann, K S Kim, D. Grab, L. B. Corbeil
Pathogenesis of Myocardial and Brain Endothelial Cell Infection by Histophilus somni
|
Progress 02/28/13 to 02/27/14
Outputs
Target Audience: Target audiences included veterinarians, scientists, biologics companies and agriculture or veterinary faculty. This was accomplished that the PD workshop in Hyattsville MD for animal health NIFA grantees, the CRWAD meetining in Chicago, Dec. 2013 and the NC1192 meeting for bovine respiratory disease research. Individual consults also occured. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? Joe Agnes, PhD and Changyou Lin, DVM, PhD are two post-doctoral scientists training in the Corbeil Lab. One pre Vet student, Ann Sung, also received training in the Corbeil lab. Heather McEligot and Nicole Behrens received training as graduate students in the Greshwin Lab. Matt Shao, a project scientist in the Gershwin lab, was involved in professional development which resulted in athe recent submission of a first author manuscript. How have the results been disseminated to communities of interest? The results have been reported at professional meetings and communicated to biologics companies and practicing veterinary pathologists. What do you plan to do during the next reporting period to accomplish the goals? Functional studies of additonal up-regulated gene products discovered in the microarray analysis are planned. Also, immunomodulation studies in vivo are planned.
Impacts
What was accomplished under these goals?
Objective 1. The hypothesis that BRSV increases sensitivity of BAT2 cells to H. somni cytotoxicity was tested in tissue culture. Cells treated with BRSV and H. somni shed virulence factors in concentrated culture supernatant (CCS) retracted more than cells treated with either agent alone. Paracellular migration of H. somni was also enhanced in dual treated cells. Objective 2. Microarray analysis of BAT2 cells treated with BRSV and CCS or either treatment alone, showed that several cytokines as well as MMPs were expressed at higher levels after dual treatment than with either agent alone. Protein ELISA assays confirmed that cytokine and MMP protein levels were both enhanced. Functional studies followed. Objective 3. Studies of immunomodulation in calves remain to be done.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Agnes JT, Zekarias B, Shao M, Anderson ML, Gershwin LJ, Corbeil LB. Bovine Respiratory Syncytial Virus and Histophilus somni Interaction at the Alveolar Barrier. Infect Immun. 81:2592-2597, 2013. PMID:23649093
|
Progress 03/01/12 to 02/27/13
Outputs
OUTPUTS: The goal was to understand mechanisms of Bacterial/viral synergy in the BRDC. We examined interaction between bovine respiratory syncytial virus and Histophilus somni at the alveolar barrier by treating bovine alveolar epithelial cells with either pathogen alone or by dual treatment. PARTICIPANTS: L B Corbeil DVM, PhD, UCSD, is the PD. L J Gershwin,DVM, PhD, UC Davis, is Co-PD. Matt Shao, PhD, UC Davis, is a Project Scientist in the Gershwin Lab. J Agnes, PhD, UCSD, is a Postdoctoral Scientist in the Corbeil Lab. TARGET AUDIENCES: Target audiences included the veterinary and agricultural scientific community, as well as biologics companies. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
To investigate BRSV/H. somni synergy in bridging the alveolar barrier, we infected bovine alveolar type 2 (BAT2) cells with BRSV for 60 hours prior to treatment with H. somni concentrated culture supernatants (CCS) for 4 hrs. Treatments included BRSV alone, H. somni CCS (enriched for IbpA) alone or dual treatment. RNA was harvested, amplified and labeled. Affymetrix Bovine Genome Arrays were used to profile gene expression patterns. Data were analyzed using Web-based software. BAT2 cell culture supernatants were collected from each treatment for measurement of selected protein production. Data were generated from three replicates. A larger number of genes were regulated by exposure to H. somni CCS than to BRSV. Gene expression analysis after dual treatment showed that 1091 genes were up- and 1279 genes down-regulated. Interestingly, some genes involved in inflammation, wound repair and tissue remodeling were synergistically up-regulated by dual challenge with BRSV and H. somni. Protein production by key up-regulated genes was determined by ELISA. Functional assays followed, to determine relevance to BRD.
Publications
- Lo KL, Kimball RA, Lehmann J, Gershwin LJ, Worby C, Corbeil LB. Antibody responses of calves to Histophilus somni recombinant IbpA subunits. Comp Immunol Microbiol Infectious Dis. 35:453-459, 2012.
|
Progress 03/01/11 to 02/28/12
Outputs
OUTPUTS: Studies of viral bacterial synergy in bovine respiratory disease were done by examining effects of BRSV, Histophilus somni or dual infection on bovine alveolar epithelial type 2 (BAT2) cells. Cytotoxicty was evaluated morphologically, crossing of the alveolar barrier was evaluated in transwell assays and gene expression was evaluated by microarray analysis. PARTICIPANTS: LB Corbeil, DVM, PhD, at UCSD and LJ Gershwin, DVM, PhD, at UC Davis participated in a multicampus collaboration in a partnership between UCSD and UC Davis. Matt Shao, PhD, is a Project Scientist in Gershwin's Lab. Bereket Zekarias, DVM, PhD, started as a Post Doctoral Fellow in Corbeil's lab and progressed to a Project Scientist, demonstrating professional development. Joe Agnes, PhD, is a post Doctoral Fellow in Corbeil's Lab as a trainee. TARGET AUDIENCES: The target audiences include veterinary scientists, veterinarians and industries involved in control of animal diseases. Efforts to reach these audiences and to influence changes in knowedge as well as actions such as development of vaccines or treatments consisted of reporting the results of studies at the Conference for Research Workers in Animal Diseases. Participants from target audiences particpate in that meeting. PROJECT MODIFICATIONS: No major changes
Impacts
To investigate BRSV/H. somni synergy in bridging the alveolar barrier, we infected bovine alveolar type 2 (BAT2) cells with BRSV for 60 hours prior to infection with H. somni. We found a significant increase in the number of retracted and rounded cells after the dual infection as compared to treatment with BRSV or H. somni alone. Transwell assays also demonstrated a significantly increased transmigration of H. somni across BAT2 monolayers infected with BRSV as compared to non-BRSV-infected BAT2 monolayers. Furthermore, we studied the genomic expression levels of BAT2 cells infected with or without BRSV for 60 hrs and then co-incubated with or without H. somni concentrated culture supernatants (CCS) for 4 hrs. RNA was harvested, amplified and labeled. Affymetrix Bovine Genome Arrays were used to profile gene expression patterns. Data were analyzed using Web-based software. BAT2 cell culture supernatants were collected from each treatment for measurement of selected protein production. Data were generated from three replicates. After BRSV infection, 188 of 24,016 transcripts exhibited significant changes in gene expression, mainly genes involved in innate and pro-inflammatory responses. Gene expression changes after exposure to H. somni CCS included 2276 genes (1176 genes up-and 1100 down-regulated). A larger number of genes were regulated by exposure to H. somni than to BRSV. In addition to the pathways induced by BRSV/H. somni, a large number of genes are involved in cell division and growth, apoptosis, and tissue repair and remodeling. Gene expression analysis after dual treatment showed that 1091 genes were up- and 1279 genes down-regulated. Interestingly, some genes involved in inflammation, wound repair and tissue remodeling were synergistically up-regulated by dual challenge with BRSV and H. somni.
Publications
- Agnes J, Zekarias B, Gershwin LJ, Corbeil LB. BRSV and H. somni synergy in bridging the alveolar barrier. CRWAD Abstract 155, 2011
- M. Shao, L.B. Corbeil, LJ. Gershwin.Genome-wide analysis of gene expression profile change in BRSV and H. somni infection of bovine alveolar epithelial cells. CRWAD Abstract 154, 2011
|
|