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Mitogen stimulation favours replication of equine herpesvirus-1 in equine blood mononuclear cells by inducing cell proliferation and formation of close intercellular contacts.
Abstract: In the present study, equine herpesvirus-1 (EHV-1)-infected cells were identified in ionomycin/phorbol dibutyrate (IONO/PDB)-stimulated peripheral blood mononuclear cells (PBMC) and the mechanism by which stimulation increases the percentage of infected cells was examined. In the population of viral antigen-positive PBMC, 38.4+/-4.5% were CD5(+) T-lymphocytes (18.1+/-3.2% CD4(+) 13.6+/-1.8% CD8(+)), 18.1+/-5.4% were B-lymphocytes, 8.5+/-3.9% were monocytes and 35% remained unidentified. The role of the cell cycle in the increased susceptibility to EHV-1 upon stimulation was examined by stimulating PBMC for 0, 12, 24 or 36 h prior to inoculation. A high correlation was found between the increase of cells in the S- (r=0.974) and G(2)/M-phase (r=0.927) at the moment of inoculation and the increase of infected cells at 12 h post-inoculation (p.i.). This suggests that a specific stage of the S-phase or S- and G(2)/M-phase facilitates virus replication. At 24 h p.i. lower correlations were found, suggesting that other effects are involved. From 12 h after addition of IONO/PDB, formation of clusters of PBMC became manifest. We examined whether close intercellular contacts in these clusters facilitated cell-to-cell transmission of EHV-1. Between 8 and 17 h p.i., the percentage of clusters containing adjacent infected cells increased from 1.6 to 13.4% and the maximal number of adjacent infected cells increased from two to four. Confocal microscopy visualized close intercellular contacts between adjacent infected cells. It can be concluded that mitogen stimulation favours EHV-1 infection of PBMC (i) by initiating specific cell cycle events and (ii) by inducing formation of clusters, thereby facilitating transmission of virus between cells.
Publication Date: 2001-07-18 PubMed ID: 11458002DOI: 10.1099/0022-1317-82-8-1951Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
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The research article investigates how mitogen stimulation can increase the replication of equine herpesvirus-1 (EHV-1) in equine blood mononuclear cells by invoking cell proliferation and promoting close intercellular contacts.
Study Background and Methodology
The researchers focused on the identification of EHV-1-infected cells in peripheral blood mononuclear cells (PBMCs) exposed to ionomycin/phorbol dibutyrate (IONO/PDB). This process served to stimulate PBMCs and investigate the mechanism behind the observed increase of infected cells post-stimulation.
- Various types of cells infected by EHV-1 were identified, including T-lymphocytes, B-lymphocytes, monocytes, and a number of unidentified cells.
Role of Cell Cycle on Susceptibility to EHV-1
The researchers further examined the role of the cell cycle in increasing susceptibility to EHV-1 upon stimulation. The PBMCs were subjected to different periods of stimulation before inoculation.
- The findings revealed a high correlation between an increase in cells entering the S- and G(2)/M-phase during inoculation and the surge of infected cells at 12 hours post-inoculation, suggesting these stages of the cell cycle could facilitate virus replication.
- At 24 hours post-inoculation, the correlations were lower, indicating that other factors may be contributing to the increase in infected cells.
Formation of Clusters of PBMCs
An observation of note was the formation of clusters of PBMCs 12 hours after addition of the mitogens IONO/PDB. The research team investigated whether these close intercellular contacts facilitated the transmission of EHV-1.
- Between 8 and 17 hours post-inoculation, there was a significant increase observed in the percentage of clusters containing adjacent infected cells and the maximal number of adjacent infected cells.
- Further visualization via confocal microscopy confirmed close intercellular contacts between adjacent infected cells.
Conclusion
The research concluded that mitogen stimulation aids in EHV-1 infection of PBMCs by:
- Initiating specific cell cycle events that appears to enhance the virus’s ability to replicate.
- Promoting the formation of cell clusters, thus facilitating the transmission of the virus between cells.
Cite This Article
APA
van der Meulen KM, Nauwynck HJ, Pensaert MB.
(2001).
Mitogen stimulation favours replication of equine herpesvirus-1 in equine blood mononuclear cells by inducing cell proliferation and formation of close intercellular contacts.
J Gen Virol, 82(Pt 8), 1951-1957.
https://doi.org/10.1099/0022-1317-82-8-1951 Publication
Researcher Affiliations
- Laboratory of Virology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium1.
- Laboratory of Virology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium1.
- Laboratory of Virology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium1.
MeSH Terms
- Animals
- B-Lymphocytes / immunology
- B-Lymphocytes / virology
- CD4-Positive T-Lymphocytes / immunology
- CD4-Positive T-Lymphocytes / virology
- CD5 Antigens / analysis
- CD8-Positive T-Lymphocytes / immunology
- CD8-Positive T-Lymphocytes / virology
- Cell Aggregation
- Cell Cycle
- Herpesviridae Infections / virology
- Herpesvirus 1, Equid / physiology
- Horses
- Ionomycin / pharmacology
- Leukocytes, Mononuclear / drug effects
- Leukocytes, Mononuclear / immunology
- Leukocytes, Mononuclear / virology
- Lymphocyte Count
- Mitogens / pharmacology
- Monocytes / immunology
- Monocytes / virology
- Phorbol Esters / pharmacology
- T-Lymphocytes / immunology
- T-Lymphocytes / virology
- Virus Replication
Citations
This article has been cited 3 times.- Kamel M, Pavulraj S, Fauler B, Mielke T, Azab W. Equid Herpesvirus-1 Exploits the Extracellular Matrix of Mononuclear Cells to Ensure Transport to Target Cells. iScience 2020 Oct 23;23(10):101615.
- Spiesschaert B, Goldenbogen B, Taferner S, Schade M, Mahmoud M, Klipp E, Osterrieder N, Azab W. Role of gB and pUS3 in Equine Herpesvirus 1 Transfer between Peripheral Blood Mononuclear Cells and Endothelial Cells: a Dynamic In Vitro Model. J Virol 2015 Dec;89(23):11899-908.
- Kumar M, Frid O, Sol A, Rouvinski A, Karniely S. Lumpy skin disease virus suppresses the antiviral response of bovine peripheral blood mononuclear cells that support viral dissemination. Vet Res 2025 Apr 26;56(1):93.
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