New methods and reagents to improve the ferret model for human influenza infections. Martel C. a , Kirkeby S. a , Aasted B. a a Copenhagen University, Faculty of Life sciences, Institute of Veterinary Pathobiology, Denmark b Panum Institute, Institute of Odontology, Denmark.
New methods and reagents to improve the ferret model for human influenza infections
Martel C. a, Kirkeby S. a, Aasted B. a
a Copenhagen University, Faculty of Life sciences, Institute of Veterinary Pathobiology, Denmark
b Panum Institute, Institute of Odontology, Denmark
Abstract:The ferret has been extensively used to study human influenza infections. However, its value as a model has suffered from the limited set of reagents and methods available for this animal. We have recently tested a large number of monoclonal antibodies cross-reacting with ferret CD markers (CD8, CD9, CD14, CD18, CD25, CD29, CD32, CD44, CD61, CD71, CD79b, CD88, CD104, CD172a and CD3) and cytokines (interferon-gamma, TNF-alpha, interleukine-4 and interleukine-8) for flow cytometry , as well as polyclonal antibodies cross-reacting with ferret immunoglobulins (IgA, IgG and IgM) for ELISA. Further improvements of the model will aim at establishing a reliable RT-PCR for ferret cytokines, as well as investigating the location of influenza receptors and viral particles in the respiratory tract via immunohistochemistry.
Figure 1: Monoclonal antibodies directed against human CD markers cross-reacting with ferret
Figure 2 : Bronchial wall from a normal ferret, stained for α gal 2-6 (A) and with periodic acid Schiff (B). C and D show a small bronchus from an infected ferret with similar stainings.
Figure 3:ELISA of the 3 purified ferret Ig preparations (IgG, IgA and IgM) using mink immunoglobulin class specific reagents. Coating of the plates was made with a commercial anti-canine IgA, anti-human IgM or absorbed anti-mink IgG.
Conclusion:This new set of reagents will prove invaluable in the study of influenza infections in the ferret. Monoclonal antibodies against cytokines and CD marker, along with class-specific Ig ELISAs will allow to take a look at finer immunological parameters, and histology/immunohistochemistry methods will help better understand the pathology of influenza virus in ferrets, and assess the effect of new vaccines in pre-clinical studies