Development of an anti-ferret CD4 monoclonal antibody for the characterisation of ferret T lymphocytes

J Immunol Methods. 2017 May:444:29-35. doi: 10.1016/j.jim.2017.02.009. Epub 2017 Feb 16.

Abstract

The ferret is an established animal model for a number of human respiratory viral infections, such as influenza virus and more recently, Ebola virus. However, a paucity of immunological reagents has hampered the study of cellular immune responses. Here we describe the development and characterisation of a novel monoclonal antibody (mAb) against the ferret CD4 antigen and the characterisation of ferret CD4 T lymphocytes. Recombinant production and purification of the ferret CD4 ectodomain soluble protein allowed hybridoma generation and the generation of a mAb (FeCD4) showing strong binding to ferret CD4 protein and lymphoid cells by flow cytometry. FeCD4 bound to its cognate antigen post-fixation with paraformaldehyde (PFA) which is routinely used to inactivate highly pathogenic viruses. We have also used FeCD4 in conjunction with other immune cell markers to characterise ferret T cells in both primary and secondary lymphoid organs. In summary, we have developed an important reagent for the study of cellular immunological responses in the ferret model of infectious disease.

Keywords: Antibody; CD4; Ferret; T cells.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology*
  • Antibodies, Monoclonal / metabolism
  • Antibody Specificity
  • CD4 Antigens / genetics
  • CD4 Antigens / immunology*
  • CD4 Antigens / metabolism
  • CD4-Positive T-Lymphocytes / drug effects
  • CD4-Positive T-Lymphocytes / immunology*
  • CD4-Positive T-Lymphocytes / metabolism
  • Cell Line
  • Cell Separation / methods
  • Concanavalin A / pharmacology
  • Enzyme-Linked Immunospot Assay
  • Ferrets / genetics
  • Ferrets / immunology*
  • Ferrets / metabolism
  • Flow Cytometry
  • Hybridomas
  • Immunity, Cellular*
  • Interferon-gamma / immunology
  • Interferon-gamma / metabolism
  • Lymphocyte Activation
  • Lymphoid Tissue / cytology
  • Lymphoid Tissue / immunology*
  • Lymphoid Tissue / metabolism
  • Phenotype
  • Protein Binding
  • Species Specificity
  • Transfection

Substances

  • Antibodies, Monoclonal
  • CD4 Antigens
  • Concanavalin A
  • Interferon-gamma