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

Authors: Layton, Daniel S.; Xiao, Xiaowen; Bentley, John D.; Lu, Louis; Stewart, Cameron R.; Bean, Andrew G. D.; Adams, Timothy E.

Source: JOURNAL OF IMMUNOLOGICAL METHODS, 444 29-35, MAY 2017

Brief summary of the paper: 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.