Cell-mediated immunity evaluation in foals infected with virulent equine herpesvirus-1 by multi-parameter flow cytometry.
Abstract: The cell-mediated immune (CMI) response of foals to virulent equine herpesvirus-1 (EHV-1) infection was evaluated by multi-parameter flow cytometry (FCM). Ten 7-8-month-old EHV-1 seronegative foals were infected intranasally with virulent EHV-1 and 10 foals served as uninfected controls. Blood samples were collected 6 and 7 weeks after infection to test for specific CMI responses to live heterologous EHV-1 recall antigen. The activation markers included major histocompatibility complex class II (MHC II), intracellular interferon gamma (IFN-gamma) and interleukin 4 (IL-4). The results from both tests were averaged before statistical analysis. Following EHV-1 stimulation, the MHC II expression index (EI) increased significantly in CD2+CD4+CD8- and CD2+CD4-CD8+ subsets of the infected group. At 4 days after incubation, the non-antigen stimulated CD2+CD4-CD8- subset of the infected group expressed a high percentage (61.1%) of MHC II. When stimulated with EHV-1, the MHC II expression declined significantly but remained at a relatively high percentage (34.4%). The IFN-gamma EI was significantly higher in infected foals in all major T cell subsets (CD2+) while only the CD2+CD4+CD8- subset showed a significant increase in intracellular IL-4 EI. The FCM results showed strong specific CMI responses to EHV-1 by all three tested parameters compared to the control group (p<0.01). The high MHC II expression in the CD2+CD4-CD8- subset suggests that this T cell subset may represent a gammadelta TCR repertoire and thereby plays an important role as antigen presenting cells in the horse, as reported in other species. Being able to simultaneously quantify the frequency of specific lymphocyte subsets and the expression of cytokines that characterize activation of lymphocytes and protective CMI by multi-parameter FCM enables evaluation of subset-specific CMI responses to EHV-1 infection. This system can be applied to measure CMI responses to other equine vaccines and pathogens.
Copyright 2010 Elsevier B.V. All rights reserved.
Publication Date: 2010-01-07 PubMed ID: 20116862DOI: 10.1016/j.vetimm.2009.12.010Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research paper undertakes an investigation into how the immune system of young horses (foals) responds to a virulent strain of equine herpesvirus-1 (EHV-1) infection, using a technique called multi-parameter flow cytometry.
Research Method
- The study comprised of a sample of 20 foals, aged between 7 and 8 months, divided into two equal groups: one subjected to intranasal infection of EHV-1, and the other serving as the control group.
- Blood samples were collected from these foals six and seven weeks after the start of the experiment. The researchers used these samples to test the foals’ cell-mediated immune (CMI) responses to a live variant of the EHV-1 antigen.
- Key indicators, or ‘activation markers’, used in the experiment included Major Histocompatibility Complex class II (MHC II), Interferon Gamma (IFN-gamma), and Interleukin 4 (IL-4) – substances or proteins that are representative of the body’s immune response.
Results and Findings
- The researchers found that, post-EHV-1 stimulation, the index of MHC II expression – a measure of the immune system’s response – increased notably in the CD2+CD4+CD8- and CD2+CD4-CD8+ T-cell subsets of the infected group. In comparison, a high percentage of MHC II was expressed by the non-antigen stimulated CD2+CD4-CD8- subset, four days after incubation.
- The expression of IFN-gamma, another activation marker, was considerably higher among the infected foals. However, only the CD2+CD4+CD8- subset demonstrated a significant surge in intracellular IL-4 expression.
- The high MHC II expression found in the CD2+CD4-CD8- subset suggested that this T-cell subset potentially represents a gammadelta T-cell receptor repertoire, indicating that this group of cells may play a significant role in the presentation of antigens in horses.
Implications of the Study
- The multi-parameter flow cytometry results reveal noticeable variations in cell-mediated immunity responses to EHV-1 infection between the infected group and the control group.
- This study establishes a potential method for evaluating subset-specific CMI responses to EHV-1 infection in foals. Furthermore, the method can be expanded for use in capturing CMI responses to other equine vaccines and pathogens.
Cite This Article
APA
Platt R, Sponseller BA, Chiang YW, Roth JA.
(2010).
Cell-mediated immunity evaluation in foals infected with virulent equine herpesvirus-1 by multi-parameter flow cytometry.
Vet Immunol Immunopathol, 135(3-4), 275-281.
https://doi.org/10.1016/j.vetimm.2009.12.010 Publication
Researcher Affiliations
- Department of Veterinary Microbiology and Preventive Medicine, Iowa State University, Ames, IA 50011, United States.
MeSH Terms
- Animals
- Base Sequence
- DNA Primers / genetics
- DNA, Viral / blood
- DNA, Viral / genetics
- Flow Cytometry
- Herpesviridae Infections / immunology
- Herpesviridae Infections / veterinary
- Herpesviridae Infections / virology
- Herpesvirus 1, Equid / genetics
- Herpesvirus 1, Equid / immunology
- Herpesvirus 1, Equid / pathogenicity
- Horse Diseases / immunology
- Horse Diseases / virology
- Horses
- Immunity, Cellular
- In Vitro Techniques
- Leukocytes, Mononuclear / immunology
- Lymphocyte Activation
- Reverse Transcriptase Polymerase Chain Reaction
- Vaccines, Attenuated / immunology
- Viral Vaccines / immunology
- Viremia / veterinary
- Viremia / virology
- Virulence
Citations
This article has been cited 3 times.- Hussen J, Schuberth HJ. Recent Advances in Camel Immunology.. Front Immunol 2020;11:614150.
- Gaashan MM, Al-Mubarak AIA, Hussen J. Leukocyte populations and their cell adhesion molecules expression in newborn dromedary camel calves.. Vet World 2020 Sep;13(9):1863-1869.
- Soboll Hussey G, Hussey SB, Wagner B, Horohov DW, Van de Walle GR, Osterrieder N, Goehring LS, Rao S, Lunn DP. Evaluation of immune responses following infection of ponies with an EHV-1 ORF1/2 deletion mutant.. Vet Res 2011 Feb 7;42(1):23.
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