Development of a monoclonal antibody against swine leukocyte antigen (SLA)-DR α chain and evaluation of SLA-DR expression in bone marrow-derived dendritic cells after PRRSV infection

Vet Immunol Immunopathol. 2019 May:211:19-24. doi: 10.1016/j.vetimm.2019.04.001. Epub 2019 Apr 11.

Abstract

Porcine reproductive and respiratory syndrome (PRRS) is one of the most common diseases in the global swine industry. PRRSV infection is highly restricted to cells of the monocyte-macrophage lineage. However, the lack of antibodies to swine monocyte-macrophage lineage markers significantly hampers PRRSV research. In this study, we have developed a monoclonal antibody against the swine leukocyte antigen (SLA)-DRα chain and confirmed its reactivity with endogenous expressed SLA-DR in a variety of cell lines and primary swine antigen-presenting cells (PAMs, PBMC and BM-DCs). Moreover, the level of SLA-DR expression after PRRSV infection were evaluated by our homemade Mab and a commercial anti-SLA-DR antibody. Based on our result, the protein level of SLA-DRα expression is increased after PRRSV infection in DC, while the mRNA of both SLA-DRα and SLA-DRβ were significantly inhibited by PRRSV replication. In conclusion, we successfully developed a MAb reactive with endogenous SLA-DR in western blotting, and this MAb could be a useful tool for further research and analysis. Moreover, the inconsistency of SLA-DR expression between protein and mRNA levels may suggest a novel role of DC played during the immune response after PRRSV infection.

Keywords: Bone marrow-derived dendritic cells; Monoclonal antibody; PRRSV; SLA-DR; Swine leukocyte antigen.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology*
  • Blotting, Western
  • Bone Marrow / immunology
  • Bone Marrow / metabolism
  • Cell Line
  • Dendritic Cells / immunology
  • Dendritic Cells / metabolism*
  • Enzyme-Linked Immunosorbent Assay
  • Flow Cytometry
  • Fluorescent Antibody Technique
  • HEK293 Cells
  • HLA-DR alpha-Chains / immunology*
  • HLA-DR alpha-Chains / metabolism
  • Humans
  • Immunoprecipitation
  • Mice
  • Mice, Inbred BALB C / immunology
  • Porcine Reproductive and Respiratory Syndrome / immunology*
  • Porcine respiratory and reproductive syndrome virus / immunology*
  • Recombinant Proteins
  • Swine / immunology

Substances

  • Antibodies, Monoclonal
  • HLA-DR alpha-Chains
  • Recombinant Proteins