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Veterinary immunology and immunopathology1999; 70(1-2); 43-54; doi: 10.1016/s0165-2427(99)00037-9

Determination of equid herpesvirus 1-specific, CD8+, cytotoxic T lymphocyte precursor frequencies in ponies.

Abstract: The frequency of antigen-specific, genetically restricted cytotoxic T lymphocyte precursors (CTLp) was measured in peripheral blood mononuclear cells (PBMC) of ponies before and after infection with equid herpesvirus 1 (EHV1). Split-well limiting dilution analysis (LDA) was developed to measure CTLp frequency using EHV1-infected 51Cr-labelled lymphoblasts as targets. Extensive characterisation showed that recombinant human interleukin-2, autologous antigen presenting cells and equine serum containing virus neutralising antibody were necessary for maturation of CTLp into effector CTL in vitro. CTLs were not induced when the equine serum (containing VN antibody) was replaced with either foetal calf serum or foetal equine serum (without VN antibody), or seronegative equine serum. CTLp frequency decreased significantly when CD8+ lymphocytes were depleted from the induction cultures. There was good inter- and intra-assay reproducibility using both fresh and recovered cryopreserved PBMC. Both EHV1 and EHV4 could be used to induce effector CTL which lysed EHV1-infected target cells. CTLp frequencies were measured in 2 groups of ponies: Group 1 consisted of two ponies (approx. 9 years old), which had multiple previous experimental infections with EHV1; Group 2 comprised five young (1-2 years) and two older (7 years) ponies which had presumed natural exposure to EHV1/EHV4 but no previous experimental infections. The results showed that CTLp frequencies were higher in the ponies of Group 1 compared with the others. Moreover, ponies with the higher CTLp frequencies were better protected against re-challenge infection with EHV1, showing reduced or absent clinical and virological signs. Consequently, measurement of EHV1-specific CTLp frequency is a potential in vitro correlate of immunity which may be useful for screening new vaccines in horses before embarking upon challenge protection studies to confirm efficacy.
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Publication Date: 1999-10-03 PubMed ID: 10507286DOI: 10.1016/s0165-2427(99)00037-9Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't
  • Research Support
  • U.S. Gov't
  • P.H.S.

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 research observed the number of antigen-specific, cytotoxic T lymphocyte precursors (CTLp) in ponies before and after infection with equine herpesvirus 1 (EHV1). It found that animals with a higher CTLp frequency had lower or nonexistent clinical and virological signs when re-infected with EHV1, suggesting that the measurement of EHV1-specific CTLp frequency may be a useful way to screen new vaccines in horses.

Methods

  • The research team developed a split-well limiting dilution analysis (LDA) to measure the frequency of CTLp. This method involved using EHV1-infected 51Cr-labelled lymphoblasts as targets.
  • Detailed characterisation indicated that recombinant human interleukin-2, as well as autologous antigen presenting cells and equine serum with virus neutralising antibody, were needed for the maturation of CTLp into effector CTL in vitro.
  • The team confirmed that CTLs were not induced when the equine serum was replaced with infected or disease-negative equine serum or foetal calf serum.
  • If CD8+ lymphocytes were removed from the induction cultures, the CTLp frequency significantly decreased.

Results

  • PBMCs, fresh and those recovered after cryopreservation, showed good inter- and intra-assay reproducibility.
  • Both EHV1 and EHV4 could induce effector CTL, which then lysed EHV1-infected target cells.
  • Two groups of ponies were tested. Group 1 had two ponies about 9 years old, which had been infected with EHV1 multiple times. Group 2 consisted of five young (1-2 years) and two older (7 years) ponies which had likely natural exposure to EHV1/EHV4 but no previous experimental infections.
  • The results showed that CTLp frequencies were higher in Group 1 ponies.
  • Ponies with higher CTLp frequencies showed reduced or absent clinical and virological signs when re-infected with EHV1.

Conclusions

  • These findings suggest that measuring EHV1-specific CTLp frequency is a potential in vitro correlate of immunity.
  • This method could be useful for screening new vaccines in horses before starting protection studies to confirm their efficiency.

Cite This Article

APA
O'Neill T, Kydd JH, Allen GP, Wattrang E, Mumford JA, Hannant D. (1999). Determination of equid herpesvirus 1-specific, CD8+, cytotoxic T lymphocyte precursor frequencies in ponies. Vet Immunol Immunopathol, 70(1-2), 43-54. https://doi.org/10.1016/s0165-2427(99)00037-9

Publication

Veterinary immunology and immunopathology
ISSN: 0165-2427
NlmUniqueID: 8002006
Country: Netherlands
Language: English
Volume: 70
Issue: 1-2
Pages: 43-54

Researcher Affiliations

O'Neill, T
  • Centre for Preventive Medicine, Animal Health Trust, Kentford, Suffolk, UK.
Kydd, J H
    Allen, G P
      Wattrang, E
        Mumford, J A
          Hannant, D

            MeSH Terms

            • Animals
            • CD8 Antigens
            • Female
            • Herpesviridae Infections / immunology
            • Herpesviridae Infections / prevention & control
            • Herpesviridae Infections / veterinary
            • Herpesvirus 1, Equid / immunology
            • Horse Diseases / immunology
            • Horse Diseases / prevention & control
            • Horses / immunology
            • Horses / virology
            • Humans
            • Male
            • T-Lymphocytes, Cytotoxic / immunology
            • T-Lymphocytes, Cytotoxic / virology
            • Viral Vaccines

            Citations

            This article has been cited 16 times.
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