Immune Response of Swine and Cattle to FMDV
Dr. Golde leads the immunology laboratory investigating the early aspects of immune responses during the acute phase of FMDV infection in swine and cattle. Among their recent discoveries is the fact that FMDV infection causes depletion of lymphocytes and therefore it disrupts the immune response against the virus. Not only that, but the virus disrupts the ability of immune cells to synthesize important molecules involved in initiating immunity, namely interferon gamma. This indicates the virus has evolved a mechanism to specifically inhibit this interferon. To more clearly understand this aspect of immune evasion by the FMD virus, they also demonstrated for the first time that porcine interferon gamma acts against FMDV, blocking virus infection in vitro. Therefore, the evolution of mechanisms to inhibit interferon production allows this virus to evade early immune responses. They have developed a method of studying skin dendritic cells of swine where 10 to 100 million enriched DC can be harvested in an overnight culture. These cells are very capable of controlling virus replication with a rapid and strong type I interferon response.
New projects include an analysis of dendritic cells from mucosal tissue to determine if these cells have the same strong innate response to virus as skin-derived DC. In addition, in collaboration with other institutions, these investigators are developing more sophisticated technologies to analyze the T cell response to FMDV including the production and testing of swine MHC tetramers. Finally, this group has developed many reagents and assays necessary to study the immune responses of cattle and swine using new technologies including adenovirus vectored expression systems. This work is fundamental to designing intervention strategies in antiviral therapy against FMD.
Lab staff includes one support scientist, one postdoctoral fellow, one visiting scientist, and a second visiting scientist co-mentored with Marvin Grubman.
Selected References:
Rodriguez, L.L., J. Barrera, E. Kramer, J. Lubroth, F. Brown and W. T. Golde. A Synthetic Peptide Containing the Consensus Sequence of the G-H Loop Region of FMDV Type O VP1 Combined with a Promiscuous T-Helper Epitope Induces Peptide-Specific Antibodies but Fails to Protect Cattle Against Viral Challenge. Vaccine. 21 (2003): 3751-3756
Bautista, E., G.S. Ferman, and W.T. Golde. Induction of Lymphopenia and Inhibition of T Cell Function During Acute Infection of Swine with Foot and Mouth Disease Virus (FMDV). Veterinary Immunology and Immunopathology. 92 (2003): 61-73
Bautista, E., D. Gregg, and W.T. Golde. Characterization and Functional Analysis of Skin-Derived Dendritic Cells from Swine without a Requirement for In Vitro Propagation. Veterinary Immunology and Immunopathology. 87 (2002): 137-145
Golde, W.T., GS Ferman, EM. Bautista, JE Butler, MCS Brum, and MJ Grubman. Immune Modulation during Acute Infection of Swine with Foot-and-Mouth Disease Virus (FMDV): Inhibition of Th1 Cytokine Production and Lack of an IgG2 Serum Antibody Response to Virus. Immunology. (Submitted)
Bautista, EM, GS Ferman, GR Gregg, and WT Golde. The Innate Immune Response to Foot-and-Mouth Disease Virus (FMDV): Constitutive Expression of IFNa and Rapid Induction of IFNß by Dendritic Cells Make them Refractory to Infection. J. Virology. (Submitted)
Outbreak Interventions (fast-acting vaccines and antivirals)
Dr. Grubman has expertise on FMDV replication and vaccine and antiviral research and has worked at PIADC for the last 28 years. Among his major contributions are the description of FMDVs mechanisms of inhibition of host response particularly interferon response. He discovered that FMDV’s lacking the 5’ leader proteinase (Lpro) grew to lower titers in vitro and were attenuated in animals due to the lack of inhibition of translation of cellular factors, including type I interferons which in turn inhibited virus growth. In recent years he showed that an adenovirus vector containing the structural proteins of FMDV and the 3C protease, could produce virus-like empty capsids that were not infectious but that could induce an immune response similar to that induced by inactivated vaccines, without inducing antibodies to non-structural viral proteins hence making vaccinated animals easily distinguishable from infected ones. Dr. Grubman has demonstrated that empty capsid vaccines are efficacious in swine against viral challenge. More recently, Dr. Grubman has shown that type I interferons delivered in adenovirus vectors can completely protect swine challenged as early as one day postadministration.
Current studies involve efforts to improve foreign gene expression in adenovirus vectors, examine virus control of type I interferon response at the molecular level, use of RNA interference to control virus replication and knock-out selected host gene targets, and examination of the early stages of virus pathogenesis.
Present staff includes one research support scientist, one postdoctoral fellow and three visiting scientists, one of whom also works with Dr. William Golde.
Selected References:
Grubman, M. J. and Baxt, B. Foot-and-Mouth Disease. Clinical Micro. Rev. 17 (2004):465-493.
Grubman, M. J. New Approaches to Rapidly Control Foot-and-Mouth Disease Outbreaks. Expert Rev. Anti-infect. Ther. 1 (2003):89-96.
Moraes, M.P., Chinsangaram, J., Brum, M.C.S., and Grubman, M.J. Immediate Protection of Swine from Foot-and-Mouth Disease: A Combination of Adenoviruses Expressing Interferon Alpha and a Foot-And-Mouth Disease Virus Subunit Vaccine. Vaccine. 22 (2003):268-279.
Wu, Q., Brum, M.C.S., Caron, L., Koster, M., and Grubman, M.J. Adenovirus-Mediated Type I Interferon Expression Delays and Reduces Disease Signs in Cattle Challenged with Foot-and-Mouth Disease Virus. J. Int. Cyt. Res., 23 (2003):371-380.
Wu, Q., Moraes, M.P., and Grubman, M.J. Recombinant Adenovirus Co-Expressing Capsid Proteins of Two Serotypes of Foot-and-Mouth Disease Virus (FMDV): In Vitro Characterization and Induction of Neutralizing Antibodies against FMDV in Swine. Virus Res. 93 (2003):211-219.
Mason, P.W., Chinsangaram, J., Moraes, M.P., Mayr, G.A., and Grubman, M.J. Engineering Better Vaccines for Foot-and-Mouth Disease. Vaccines for OIE List A and Emerging Animal Diseases. Dev. Biol. Basel, Karger (Brown, F, Roth, J, eds). 114 (2003):79-88.
Grubman, M.J., Moraes, M.P., Chinsangaram, J., Mayr, G.A., and Mason, P.W. New Approaches to Control Foot-and-Mouth Disease. Foot and Mouth Disease: Control Strategies. (B. Dodet and M. Vicari eds). Elsevier. (2003). p. 337-343.
Chinsangaram, J., Moraes, M.P., Koster, M. and Grubman, M.J. A Novel Viral Disease Control Strategy: Adenovirus Expressing Interferon Alpha Rapidly Protects Swine from Foot-and-Mouth Disease. J. Virol. 77 (2003):1621-1625.
Mason, P.W., Grubman, M.J., and Baxt, B. Molecular Basis of Pathogenesis of FMDV. In Foot-and-Mouth Disease (D. Rowlands ed.). Virus Res. 91 (2003):9-32.
Grubman, M.J. and Mason, P.W. Prospects, Including Time Frames for Improved Foot-and-Mouth Disease Vaccines. In Foot and Mouth Disease: Facing the New Dilemmas. Rev. sci. tech. Off. Int. Epiz. 21 (2002):589-600.
Moraes, M.P., Mayr, G.A., Mason, P.W., and Grubman, M.J. Early Protection against Homologous Challenge after a Single Dose of Replication-Defective Human Adenovirus Type 5 Expressing Capsid Proteins of Foot-and-Mouth Disease Virus (FMDV) Strain A24. Vaccine. 20 (2002):1631-1639.
Moraes, M.P., Mayr, G.A., and Grubman, M.J. pAd5-Blue: An Easy to Use, Efficient, Direct Ligation System for Engineering Recombinant Adenovirus Constructs. BioTechniques. 31 (2001):1050-1056.
Chinsangaram, J., Koster, M., and Grubman, M.J. Inhibition of L-deleted Foot-and-Mouth Disease Virus Replication by Alpha/Beta Interferon Involves Double-Stranded RNA-Dependent Protein Kinase. J. Virol. 12 (2001):5498-5503.
Mayr, G.S., O’Donnell, V., Chinsangaram, J., Mason, P.W., and Grubman, M.J. Immune Responses and Protection against Foot-and-Mouth Disease Virus (FMDV) Challenge in Swine Vaccinated with Adenovirus-FMDV Constructs. Vaccine 19 (2001):2152-2162.
Molecular Virology of FMDV
Dr. Rieder leads the molecular biology laboratory investigating FMDV proliferation and pathogenicity. Specifically, her laboratory is involved in the analysis of the basis of the pathogenic characteristics of FMDV. Using the knowledge of molecular virology via manipulation of infectious cDNA clones for FMDV and expertise in virus replication, she is identifying both viral factors (proteins and genetic elements), and host factors, that might influence how the virus causes disease. In addition, she has recently developed a cell-free system in which FMDV can be synthesized de novo to examine mechanism of virus replication and the effect of anti-viral compounds. This involves using non-replicating viral genomes, called replicons, which can be used in laboratories outside of biocontainment, meaning they would be appropriate for use in pharmaceutical companies developing anti-viral therapeutics.
The lab currently has one research support scientist.
Selected References:
Baxt, B., and Rieder, E. Molecular Aspects of Foot-and-Mouth Disease Virus Virulence and Host Range: Role of Host Cell Receptors and Viral Factors. In Sobrino, F., and Domingo, E. (eds.). Foot-and-Mouth Disease: Current Perspectives. Horizon Scientific Press, in press, (2004).
Yin J., Paul, A.V., Wimmer, E., and Rieder E. Functional Dissection of a Poliovirus Cis-Acting Element [PV-Cre(2C)]: Analysis of a Single- and Dual-Cre Viral Genomes and Protein that Bind Specifically to PV-Cre RNA. J. Virol. 77 (2003):5152-5166.
Rieder, E., Xiang W., Paul, A.V., and Wimmer, E. Analysis of the Cloverleaf Element in a Human Rhinovirus Type 14/Poliovirus Chimera: Correlation of Sub-Domain D Structure, Ternary Protein Complex Formation and Virus Replication. J. Gen.Virol. 84 (2003):2203-2216.
Rieder, E., A. Berinstein, B. Baxt, A. Kang, and P.W. Mason. Propagation of an Attenuated Virus by Design: Engineering a Novel Receptor for Non-Infectious Foot-and-Mouth Disease Virus. Proc. Natl. Acad. Sci. USA 93 (1996): 10428-10433.
Rieder, E., B. Baxt, and P.W. Mason. Animal-Derived Antigenic Variants of FMDV Type A12 Have Low Affinity for Cells in Culture. J. Virol. 68 (1994): 5296-5299.
Vesicular stomatitis virus genomics, molecular epidemiology and pathogenesis
This project has two components both designed to understand and minimize the occurrence of vesicular stomatitis in the U.S. The first component addresses animal pathogenesis aspects, specifically the role host factors (e.g. local – non-specific immunity) and of specific viral genes in determining the outcome of viral infections. This research could result in the development of therapeutic agents capable of preventing clinical disease during outbreaks not only of VS but also of FMD. The second component utilizes viral genomics for diagnosis and molecular epidemiology, to rapidly detect, identify and track the origin of VSV strains causing outbreaks. Phylogenetic analyses are utilized to understand patterns of viral evolution in nature and the environmental factors influencing the occurrence of the disease.
Selected References:
Martinez, I. Barrera, J., Rodriguez, L.L., Wertz, G.W. Recombinant Vesicular Stomatitis (Indiana) Virus Expressing New Jersey and Indiana Glycoproteins Induces Neutralizing Antibodies to each Serotype in Swine, a Natural Host. Vaccine (2004); In Press.
Martinez, I., Rodriguez, L.L., Jimenez, C., Pauszek, S.J., Wertz, G.W. Vesicular Stomatitis Virus Glycoprotein is a Determinant of Pathogenesis in Swine, a Natural Host. J.Virol. 77 (2003): 8039-8047.
Llewellyn, Z.N., Salman, M.D., Pauszek, S., Rodriguez, L.L. Growth and molecular evolution of vesicular stomatitis serotype New Jersey in cells derived from its natural insect-host: evidence for natural adaptation. Virus Res. (2002); 89: 65-73.
Rodriguez, L.L., Pauszek, S.J., Bunch, T.A., Schumann, K.R. Full-Length Genome Analysis of Natural Isolates of Vesicular Stomatitis Virus (Indiana 1 Serotype) from North, Central and South America. J.Gen.Virol. 83 (2002): 2475-2483.
Rodriguez, L. L. Emergence and Re-Emergence of Vesicular Stomatitis in the United States. Virus Res. 85 (2002): 211-219.
Flanagan, E.B., Zamparo, J. M., Ball, L.A., Rodriguez, L.L., Wertz, G.W. Rearrangement of the Genes of Vesicular Stomatitis Virus Eliminates Clinical Disease in the Natural Host: New Strategy for Vaccine Development. J.Virol. 75 (2001): 6107-6114.
Rodriguez, L.L., Bunch, T.A., Fraire M., Llewellyn, Z.N. Re-Emergence of Vesicular Stomatitis in the Western United States is Associated with Distinct Viral Genetic Lineages. Virology. 271 (2000): 171-181.