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Home / Equine Research Bank / Vet Immunol Immunopathol / The determination of in vivo envelope-specific cell-mediated...
Veterinary immunology and immunopathology2012; 148(3-4); 302-310; doi: 10.1016/j.vetimm.2012.06.018

The determination of in vivo envelope-specific cell-mediated immune responses in equine infectious anemia virus-infected ponies.

Abstract: Distinct from human lentivirus infection, equine infectious anemia virus (EIAV)-infected horses will eventually enter an inapparent carrier state in which virus replication is apparently controlled by adaptive immune responses. Although recrudescence of disease can occur after immune suppression, the actual immune correlate associated with protection has yet to be determined. Therefore, EIAV provides a model for investigating immune-mediated protective mechanisms against lentivirus infection. Here, we have developed a method to monitor EIAV-envelope specific cellular immunity in vivo. An EIA carrier horse with no clinical signs infected 7 years ago and 4 related experimental ponies infected 6 months previously were used in this study. Forty-four 20-mer peptides, representing the entire surface unit protein (gp90) of EIAV, were combined into 14 peptide pools and intradermally injected into the neck of EIAV-infected horses. An identical volume of saline alone was injected into a fifteenth site as a negative control. After 48 h, those sites with palpable infiltrations were measured prior to the collection of 2mm and 4mm punch biopsies. Total RNA was extracted from each 2mm biopsy for determination of CD3 and interferon-γ (IFN-γ) mRNA expression by real-time PCR. The 4mm skin biopsies were formalin-fixed and paraffin-embedded for immunohistochemistry (IHC) staining for CD3, CD20, CD25 and MAC387 (macrophage marker). Peripheral blood mononuclear cells (PBMC) were obtained prior to the injection and tested for in vitro reactivity against the same peptides. Histological examination showed that some of the envelope peptides elicited a lymphocytic cellular infiltration at the injection site, as evidenced by positive staining for CD3. Gp90 peptide-specific increases in CD3 and IFN-γ gene expression were also detected in the injection sites. Furthermore, differences were found between in vivo and in vitro responses to gp90 specific peptides. These results demonstrate a novel method for detecting in vivo cell-mediated immune responses to EIAV-specific peptides that is readily applicable to other host/pathogen systems.
Copyright © 2012 Elsevier B.V. All rights reserved.
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Publication Date: 2012-06-23 PubMed ID: 22795699PubMed Central: PMC4337896DOI: 10.1016/j.vetimm.2012.06.018Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This study involves creating a new method to monitor the immune responses exhibited by horses infected with the equine infectious anemia virus (EIAV), a lentivirus. The study particularly focuses on understanding these immune responses at a cellular level in a living organism (in vivo), facilitating further understanding of protective mechanisms against lentivirus infections.

Research Context

  • Equine infectious anemia virus (EIAV) is a lentivirus similar to those causing chronic illness in humans. The peculiarity of EIAV lies in its carriage by infected horses, who, unlike humans, eventually reach a state where the virus replication is controlled by their adaptive immune responses.
  • Understanding these immune responses could provide insights into how a similar control mechanism could be achieved in human lentivirus infections. Hence, the EIAV case serves as an excellent model for lentivirus study.

Methodology

  • The research involved an EIA carrier horse and four experimentally infected ponies with no observable clinical signs of illness.
  • All the subjects were intradermally exposed to 14 peptide pools containing 44 peptides, representing the complete surface unit protein (gp90) of EIAV. A fifteenth control injection containing only saline was also administered.
  • After 48 hours, injection sites with discernible infiltration were measured, upon which 2mm and 4mm skin biopsies were taken. The expression of CD3 and interferon-γ (IFN-γ) mRNA was subsequently determined using a real-time PCR.
  • The larger skin biopsies were also processed for immunohistochemical staining to investigate several immune cellular markers such as CD3, CD20, CD25, and MAC387.

Study Results

  • Findings reveal that some peptides are successful in eliciting a lymphocytic cellular infiltration, as evidenced through a positive CD3 stain. This demonstrates a specific increase in the CD3 and IFN-γ gene expression, indicative of an immune response to the EIAV-specific peptides.
  • Interestingly, the study suggests that the immune responses detected in vivo differ from those discerned through in vitro techniques.

Conclusion

  • The team successfully developed a novel method to detect the nature of immune responses to specific lentivirus peptides in a living host, thereby enhancing our understanding of lentivirus, and more specifically, EIAV infections.
  • This advance paves the way for further in-depth studies and potential future applications in other host/pathogen investigations.

Cite This Article

APA
Liu C, Cook FR, Cook SJ, Craigo JK, Even DL, Issel CJ, Montelaro RC, Horohov DW. (2012). The determination of in vivo envelope-specific cell-mediated immune responses in equine infectious anemia virus-infected ponies. Vet Immunol Immunopathol, 148(3-4), 302-310. https://doi.org/10.1016/j.vetimm.2012.06.018

Publication

Veterinary immunology and immunopathology
ISSN: 1873-2534
NlmUniqueID: 8002006
Country: Netherlands
Language: English
Volume: 148
Issue: 3-4
Pages: 302-310

Researcher Affiliations

Liu, Chong
  • Maxwell H. Gluck Equine Research Center, Department of Veterinary Science, University of Kentucky, Lexington, KY 40546, USA.
Cook, Frank R
    Cook, Sheila J
      Craigo, Jodi K
        Even, Deborah L
          Issel, Charles J
            Montelaro, Ronald C
              Horohov, David W

                MeSH Terms

                • Animals
                • Carrier State / immunology
                • Carrier State / virology
                • Eosine Yellowish-(YS)
                • Equine Infectious Anemia / immunology
                • Gene Expression Regulation, Viral / immunology
                • Hematoxylin
                • Horses / immunology
                • Horses / virology
                • Immunity, Cellular / immunology
                • Infectious Anemia Virus, Equine / immunology
                • Real-Time Polymerase Chain Reaction / veterinary
                • Viral Envelope Proteins / immunology
                • Viral Envelope Proteins / metabolism

                Grant Funding

                • R01 AI025850 / NIAID NIH HHS
                • UL1 TR000005 / NCATS NIH HHS
                • R01 AI25850-23 / NIAID NIH HHS

                Conflict of Interest Statement

                . The authors declare that no conflict of interest exists.

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                Citations

                This article has been cited 3 times.
                1. Carossino M, Loynachan AT, Canisso IF, Cook RF, Campos JR, Nam B, Go YY, Squires EL, Troedsson MHT, Swerczek T, Del Piero F, Bailey E, Timoney PJ, Balasuriya UBR. Equine Arteritis Virus Has Specific Tropism for Stromal Cells and CD8(+) T and CD21(+) B Lymphocytes but Not for Glandular Epithelium at the Primary Site of Persistent Infection in the Stallion Reproductive Tract.. J Virol 2017 Jul 1;91(13).
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                2. Craigo JK, Ezzelarab C, Cook SJ, Liu C, Horohov D, Issel CJ, Montelaro RC. Protective efficacy of centralized and polyvalent envelope immunogens in an attenuated equine lentivirus vaccine.. PLoS Pathog 2015 Jan;11(1):e1004610.
                  doi: 10.1371/journal.ppat.1004610pubmed: 25569288google scholar: lookup
                3. Liu C, Cook SJ, Craigo JK, Cook FR, Issel CJ, Montelaro RC, Horohov DW. Epitope shifting of gp90-specific cellular immune responses in EIAV-infected ponies.. Vet Immunol Immunopathol 2014 Oct 15;161(3-4):161-9.
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