title: Structural and functional characterisation of ferret interleukin-2.
abstract: While the ferret is a valuable animal model for a number of human viral infections, such as influenza, Hendra and Nipah, evaluating the cellular immune response following infection has been hampered by the lack of a number of species-specific immunological reagents. Interleukin 2 (IL-2) is one such key cytokine. Ferret recombinant IL-2 incorporating a C-terminal histidine tag was expressed and purified and the three-dimensional structure solved and refined at 1.89 Å by X-ray crystallography, which represents the highest resolution and first non-human IL-2 structure. While ferret IL-2 displays the classic cytokine fold of the four-helix bundle structure, conformational flexibility was observed at the second helix and its neighbouring region in the bundle, which may result in the disruption of the spatial arrangement of residues involved in receptor binding interactions, implicating subtle differences between ferret and human IL-2 when initiating biological functions. Ferret recombinant IL-2 stimulated the proliferation of ferret lymph node cells and induced the expression of mRNA for IFN-γ and Granzyme A.
authors.affiliation: CSIRO Manufacturing, Parkville, VIC 3052, Australia.
authors.forename: Bin
authors.initials: B
authors.lastname: Ren
authors.affiliation: CSIRO Manufacturing, Parkville, VIC 3052, Australia.
authors.forename: William J
authors.initials: WJ
authors.lastname: McKinstry
authors.affiliation: CSIRO Manufacturing, Parkville, VIC 3052, Australia.
authors.forename: Tam
authors.initials: T
authors.lastname: Pham
authors.affiliation: CSIRO Manufacturing, Parkville, VIC 3052, Australia.
authors.forename: Janet
authors.initials: J
authors.lastname: Newman
authors.affiliation: CSIRO Health and Biosecurity, Geelong, VIC 3219, Australia.
authors.forename: Daniel S
authors.initials: DS
authors.lastname: Layton
authors.affiliation: CSIRO Health and Biosecurity, Geelong, VIC 3219, Australia.
authors.forename: Andrew G
authors.initials: AG
authors.lastname: Bean
authors.affiliation: Department of Microbiology and Immunology, The University of Melbourne at the Doherty Institute, Melbourne, VIC 3000, Australia.
authors.forename: Zhenjun
authors.initials: Z
authors.lastname: Chen
authors.affiliation: WHO Collaborating Centre for Reference and Research on Influenza (VIDRL), Peter Doherty Institute for Infection & Immunity, Melbourne, Australia.
authors.forename: Karen L
authors.initials: KL
authors.lastname: Laurie
authors.affiliation: WHO Collaborating Centre for Reference and Research on Influenza (VIDRL), Peter Doherty Institute for Infection & Immunity, Melbourne, Australia.
authors.forename: Kathryn
authors.initials: K
authors.lastname: Borg
authors.affiliation: WHO Collaborating Centre for Reference and Research on Influenza (VIDRL), Peter Doherty Institute for Infection & Immunity, Melbourne, Australia.
authors.forename: Ian G
authors.initials: IG
authors.lastname: Barr
authors.affiliation: CSIRO Manufacturing, Parkville, VIC 3052, Australia. Electronic address: Tim.Adams@csiro.au.
authors.forename: Timothy E
authors.initials: TE
authors.lastname: Adams
mesh_terms: D000818:Animals; D049109:Cell Proliferation; D002478:Cells, Cultured; D018360:Crystallography, X-Ray; D005289:Ferrets / Q000276:immunology*; D053804:Granzymes / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D007376:Interleukin-2 / Q000235:genetics / Q000378:metabolism*; D008198:Lymph Nodes / Q000276:immunology* / Q000473:pathology; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000235:genetics; D013045:Species Specificity; D040681:Structural Homology, Protein; D014777:Virus Diseases / Q000276:immunology*
chemical_list: D007376:Interleukin-2; D011994:Recombinant Proteins; D007371:Interferon-gamma; D053804:Granzymes
keywords: Ferret; Lymphocyte proliferation; Recombinant interleukin-2; X-ray structure; mRNA induction