title: Development of a novel, guinea pig-specific IFN-γ ELISPOT assay and characterization of guinea pig cytomegalovirus GP83-specific cellular immune responses following immunization with a modified vaccinia virus Ankara (MVA)-vectored GP83 vaccine.
abstract: The guinea pig (Cavia porcellus) provides a useful animal model for studying the pathogenesis of many infectious diseases, and for preclinical evaluation of vaccines. However, guinea pig models are limited by the lack of immunological reagents required for characterization and quantification of antigen-specific T cell responses. To address this deficiency, an enzyme-linked immunospot (ELISPOT) assay for guinea pig interferon (IFN)-γ was developed to measure antigen/epitope-specific T cell responses to guinea pig cytomegalovirus (GPCMV) vaccines. Using splenocytes harvested from animals vaccinated with a modified vaccinia virus Ankara (MVA) vector encoding the GPCMV GP83 (homolog of human CMV pp65 [gpUL83]) protein, we were able to enumerate and map antigen-specific responses, both in vaccinated as well as GPCMV-infected animals, using a panel of GP83-specific peptides. Several potential immunodominant GP83-specific peptides were identified, including one epitope, LGIVHFFDN, that was noted in all guinea pigs that had a detectable CD8+ response to GP83. Development of a guinea pig IFN-γ ELISPOT should be useful in characterization of additional T cell-specific responses to GPCMV, as well as other pathogens. This information in turn can help focus future experimental evaluation of immunization strategies, both for GPCMV as well as for other vaccine-preventable illnesses studied in the guinea pig model.
authors.affiliation: University of Minnesota Medical School, Department of Pediatrics, Center for Infectious Diseases and Microbiology Translational Research, 2001 6th Street SE, Minneapolis, MN 55455, United States. Electronic address: gill0221@umn.edu.
authors.forename: Peter A
authors.initials: PA
authors.lastname: Gillis
authors.affiliation: University of Minnesota Medical School, Department of Pediatrics, Center for Infectious Diseases and Microbiology Translational Research, 2001 6th Street SE, Minneapolis, MN 55455, United States. Electronic address: hernande@umn.edu.
authors.forename: Nelmary
authors.initials: N
authors.lastname: Hernandez-Alvarado
authors.affiliation: University of Minnesota Medical School, Department of Pediatrics, Center for Infectious Diseases and Microbiology Translational Research, 2001 6th Street SE, Minneapolis, MN 55455, United States. Electronic address: gnan0007@gmail.com.
authors.forename: Josephine S
authors.initials: JS
authors.lastname: Gnanandarajah
authors.affiliation: Department of Virology, Beckman Research Institute of City of Hope, 1500 E. Duarte Road, Duarte, CA 91010, United States. Electronic address: fwussow@coh.org.
authors.forename: Felix
authors.initials: F
authors.lastname: Wussow
authors.affiliation: Department of Virology, Beckman Research Institute of City of Hope, 1500 E. Duarte Road, Duarte, CA 91010, United States. Electronic address: DDiamond@coh.org.
authors.forename: Don J
authors.initials: DJ
authors.lastname: Diamond
authors.affiliation: University of Minnesota Medical School, Department of Pediatrics, Center for Infectious Diseases and Microbiology Translational Research, 2001 6th Street SE, Minneapolis, MN 55455, United States. Electronic address: schleiss@umn.edu.
authors.forename: Mark R
authors.initials: MR
authors.lastname: Schleiss
mesh_terms: D000818:Animals; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D003586:Cytomegalovirus Infections / Q000517:prevention & control*; D004195:Disease Models, Animal; D058501:Enzyme-Linked Immunospot Assay; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D006168:Guinea Pigs; D007111:Immunity, Cellular; D007371:Interferon-gamma / Q000276:immunology; D019211:Roseolovirus / Q000276:immunology*; D013154:Spleen / Q000166:cytology / Q000276:immunology; D014616:Vaccinia virus; D014763:Viral Matrix Proteins / Q000276:immunology*; D014765:Viral Vaccines / Q000276:immunology*
chemical_list: D018984:Epitopes, T-Lymphocyte; D014763:Viral Matrix Proteins; D014765:Viral Vaccines; D007371:Interferon-gamma
keywords: CMV pp65; Congenital CMV infection; Cytomegalovirus vaccine; ELISPOT; Glycoprotein B; Guinea pig; Guinea pig cytokine; Guinea pig immunology; IFN-γ; Interferon gamma; Placenta; TORCH infection; pp65