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Identification of GPCMV infected cells in vitro and in vivo with a monoclonal antibody
C T Jones1, S K Keay, P T Swoveland
1Department of Pathology, University of Maryland, School of Medicine, Baltimore 21201.
Abstract:
A monoclonal antibody was made which identifies a 160-180 kDa structural protein in guinea pig cytomegalovirus (GPCMV) infected cells by Western blot using non-reducing conditions. This protein was shown to be a virion structural protein by purification of GPCMV on a density viscosity gradient and Western blot analysis. Phosphoanacetic acid (PAA) experiments suggest that the protein is a late GPCMV protein. In vitro the monoclonal antibody labels a cytoplasmic protein in infected guinea pig embryo fibroblasts by 12 h postinfection. The monoclonal antibody also identifies GPCMV infected cells in vivo in paraffin embedded formalin fixed tissue.
Insights
A new monoclonal antibody identifies a key 160-180 kDa structural protein in guinea pig cytomegalovirus (GPCMV) infected cells. This antibody also detects GPCMV in tissues, aiding in viral diagnostics.
Area of Science:
- Virology
- Immunology
- Cell Biology
Background:
- Guinea pig cytomegalovirus (GPCMV) is a significant pathogen.
- Understanding GPCMV structural proteins is crucial for developing diagnostic and therapeutic strategies.
Purpose of the Study:
- To develop and characterize a monoclonal antibody targeting a specific GPCMV structural protein.
- To investigate the properties and localization of this identified protein.
Main Methods:
- Western blot analysis under non-reducing conditions.
- Purification of GPCMV virions using density-viscosity gradients.
- Immunofluorescence staining of infected cells.
- Analysis of infected guinea pig tissues.
Main Results:
- A monoclonal antibody was generated that recognizes a 160-180 kDa protein in GPCMV-infected cells.
- This protein was confirmed as a virion structural component.
- Phosphonoacetic acid (PAA) experiments indicated the protein is expressed late in the viral replication cycle.
- The antibody labeled a cytoplasmic protein in infected fibroblasts and detected GPCMV in fixed tissues.
Conclusions:
- A novel monoclonal antibody identifies a late, virion-associated structural protein of GPCMV.
- This antibody is a valuable tool for detecting GPCMV infection in vitro and in vivo.
- Further characterization of this protein may reveal its role in viral pathogenesis.