Porcine γδ T cells express cytotoxic cell-associated markers and display killing activity but are not selectively cytotoxic against PRRSV- or swIAV-infected macrophages

Front Immunol. 2024 Jul 31:15:1434011. doi: 10.3389/fimmu.2024.1434011. eCollection 2024.

Abstract

Background: Gamma-delta (γδ) T cells are a major immune cell subset in pigs. Approximately 50% of circulating T cells are γδ T cells in young pigs and up to 30% in adult sows. Despite this abundance, the functions of porcine γδ T cells are mostly unidentified. In humans and mice, activated γδ T cells exhibit broad innate cytotoxic activity against a wide variety of stressed, infected, and cancerous cells through death receptor/ligand-dependent and perforin/granzyme-dependent pathways. However, so far, it is unknown whether porcine γδ T cells have the ability to perform cytotoxic functions.

Methods: In this study, we conducted a comprehensive phenotypic characterization of porcine γδ T cells isolated from blood, lung, and nasal mucosa. To further analyze the cytolytic potential of γδ T cells, in vitro cytotoxicity assays were performed using purified γδ T cells as effector cells and virus-exposed or mock-treated primary porcine alveolar macrophages as target cells.

Results: Our results show that only CD2+ γδ T cells express cytotoxic markers (CD16, NKp46, perforin) with higher perforin and NKp46 expression in γδ T cells isolated from lung and nasal mucosa. Moreover, we found that γδ T cells can exhibit cytotoxic functions in a cell-cell contact and degranulation-dependent manner. However, porcine γδ T cells did not seem to specifically target Porcine Reproductive and Respiratory Syndrome Virus or swine Influenza A Virus-infected macrophages, which may be due to viral escape mechanisms.

Conclusion: Porcine γδ T cells express cytotoxic markers and can exhibit cytotoxic activity in vitro. The specific mechanisms by which porcine γδ T cells recognize target cells are not fully understood but may involve the detection of cellular stress signals.

Keywords: Influenza A Virus; PAMs; PRRSV; Porcine γδ T cells; cytotoxicity; phenotype.

MeSH terms

  • Animals
  • Biomarkers
  • Cells, Cultured
  • Cytotoxicity, Immunologic*
  • Intraepithelial Lymphocytes / immunology
  • Macrophages, Alveolar / immunology
  • Macrophages, Alveolar / virology
  • Orthomyxoviridae Infections / immunology
  • Perforin / immunology
  • Perforin / metabolism
  • Porcine Reproductive and Respiratory Syndrome / immunology
  • Porcine respiratory and reproductive syndrome virus* / immunology
  • Porcine respiratory and reproductive syndrome virus* / physiology
  • Receptors, Antigen, T-Cell, gamma-delta / immunology
  • Receptors, Antigen, T-Cell, gamma-delta / metabolism
  • Swine
  • T-Lymphocytes, Cytotoxic / immunology

Substances

  • Receptors, Antigen, T-Cell, gamma-delta
  • Biomarkers
  • Perforin

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This research was funded by the Saskatchewan Agriculture Development Fund. LB is the recipient of the 2021-2023 Doctoral Scholarship from the German Academic Scholarship Foundation (Studienstiftung des deutschen Volkes) and 2023/2024 Student Support Fund from the Department of Veterinary Microbiology. VIDO also receives operational funding from the Government of Saskatchewan through Innovation Saskatchewan and the Ministry of Agriculture and from the Canada Foundation for Innovation through the Major Science Initiatives. Published as VIDO manuscript series no. 1059.