title: Development of an anti-ferret CD4 monoclonal antibody for the characterisation of ferret T lymphocytes.
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.
authors.affiliation: CSIRO Health and Biosecurity, Australian Animal Health Laboratory, Geelong, Victoria, Australia. Electronic address: daniel.layton@csiro.au.
authors.forename: Daniel S
authors.initials: DS
authors.lastname: Layton
authors.affiliation: CSIRO Manufacturing, Parkville, Victoria, Australia.
authors.forename: Xiaowen
authors.initials: X
authors.lastname: Xiao
authors.affiliation: CSIRO Manufacturing, Parkville, Victoria, Australia.
authors.forename: John D
authors.initials: JD
authors.lastname: Bentley
authors.affiliation: CSIRO Manufacturing, Parkville, Victoria, Australia.
authors.forename: Louis
authors.initials: L
authors.lastname: Lu
authors.affiliation: CSIRO Health and Biosecurity, Australian Animal Health Laboratory, Geelong, Victoria, Australia.
authors.forename: Cameron R
authors.initials: CR
authors.lastname: Stewart
authors.affiliation: CSIRO Health and Biosecurity, Australian Animal Health Laboratory, Geelong, Victoria, Australia.
authors.forename: Andrew G D
authors.initials: AG
authors.lastname: Bean
authors.affiliation: CSIRO Manufacturing, Parkville, Victoria, Australia.
authors.forename: Timothy E
authors.initials: TE
authors.lastname: Adams
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism; D000918:Antibody Specificity; D015704:CD4 Antigens / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D015496:CD4-Positive T-Lymphocytes / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D002460:Cell Line; D002469:Cell Separation / Q000379:methods; D003208:Concanavalin A / Q000494:pharmacology; D058501:Enzyme-Linked Immunospot Assay; D005289:Ferrets / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D005434:Flow Cytometry; D006825:Hybridomas; D007111:Immunity, Cellular; D007371:Interferon-gamma / Q000276:immunology / Q000378:metabolism; D008213:Lymphocyte Activation; D008221:Lymphoid Tissue / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D010641:Phenotype; D011485:Protein Binding; D013045:Species Specificity; D014162:Transfection
chemical_list: D000911:Antibodies, Monoclonal; D015704:CD4 Antigens; D003208:Concanavalin A; D007371:Interferon-gamma
keywords: Antibody; CD4; Ferret; T cells