Intracellular localization and epitope mapping of feline herpesvirus type 1 glycoproteins
J A Limcumpao1, T Horimoto, E Takahashi
1Department of Veterinary Microbiology, Faculty of Agriculture, University of Tokyo, Japan.
Insights
Monoclonal antibodies revealed distinct intracellular transport patterns for feline herpesvirus type 1 glycoproteins. These studies characterize feline herpesvirus type 1 (FHV-1) glycoproteins, aiding in understanding viral protein expression and localization.
Area of Science:
- Virology
- Immunology
- Cell Biology
Background:
- Feline herpesvirus type 1 (FHV-1) is a significant pathogen in cats.
- Understanding the structure and function of FHV-1 glycoproteins is crucial for developing antiviral strategies.
- Monoclonal antibodies (MoAbs) are valuable tools for characterizing viral proteins.
Purpose of the Study:
- To characterize feline herpesvirus type 1 (FHV-1) glycoproteins using monoclonal antibodies.
- To investigate the intracellular localization and transport of FHV-1 glycoproteins.
- To map the antigenic domains and epitopes of FHV-1 glycoproteins.
Main Methods:
- Sequential indirect immunofluorescence assay (IFA) on fixed and unfixed infected cells.
- Enzyme-linked immunosorbent assay (ELISA) additivity tests for epitope mapping.
Main Results:
- Distinct intracellular localization and transport patterns were observed for FHV-1 glycoproteins gp143/108, gp113, and gp60.
- All three glycoproteins were found to be expressed on the cell surface membrane.
- gp113 possesses two antigenic domains (neutralizing and non-neutralizing), while gp143/108 has one antigenic site with a conserved region.
Conclusions:
- The study elucidates the differential intracellular trafficking of key FHV-1 glycoproteins.
- Epitope mapping provides insights into the antigenic structure of FHV-1 glycoproteins, particularly gp113 and gp143/108.
- These findings contribute to a better understanding of FHV-1 pathogenesis and potential targets for immunotherapy.
Abstract:
Monoclonal antibodies (MoAbs) were used to characterize feline herpesvirus type 1 (FHV-1) glycoproteins (gp). Intracellular localization and transport of these proteins as revealed by a sequential indirect immunofluorescence assay (IFA) on fixed infected cells showed slight differences between FHV-1 gp143/108 and gp113. Antibodies against gp143/108 first showed membrane fluorescence at 4 hrs post-infection (PI) followed by a pronounced perinuclear and cytoplasmic staining from 8 hrs PI onwards. Those reacting with gp113 showed the same pattern but fluorescence did not appear until 8 hrs PI. In contrast, MoAbs against gp60 first showed para- and perinuclear staining at 12 hrs PI which became intranuclear at 16 hrs PI, followed by intracytoplasmic staining at 20 hrs PI. Sequential IFA of unfixed infected cells revealed that the three glycoproteins were expressed on the cell surface membrane as well. Topographical mapping of the functional epitopes of gp113 by ELISA additivity test indicated the presence of 2 antigenic domains--a neutralizing domain consisting of 3 overlapping epitopes and a non-neutralizing domain. On the other hand, gp143/108 contained only one antigenic site consisting of 5 similar or overlapping epitopes, one of which seemed to be a conserved region recognized by all MoAbs reacting to this protein.
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