title: In vitro functional characterization of feline IgGs.
abstract: Very little is known about the functional properties of feline IgGs. Here we report the in vitro characterization of cloned feline IgGs. Rapid amplification of cDNA ends (RACE) and full-length PCR of cat splenic cDNA were used to identify feline sequences encoding IgG heavy chain constant regions (IGHC). Two of the sequences are possibly allelic and have been previously reported in the literature as the only feline IgG, IgG1. Although we confirmed these alleles to be highly abundant (∼98%), analysis of numerous amplification products revealed an additional sequence (∼2%). We cloned and characterized chimeric monoclonal antibodies with each of these heavy chains. Using RACE we revealed the sequences for feline Fc gamma receptor I (FcγRI) and feline Fc neonatal receptor (FcRn). We constructed these recombinant receptors as well as fFcγRIII and determined their binding affinities to the chimeras. All of the chimeras bound to Protein A but not to Protein G, and bound tightly to fFcRn (KD=2-5 nM). Both IgG1 alleles have a high affinity for fFcγRI (KD=10-20 nM), they bind to the low-affinity fFcγRIII receptor (2-4 μM), and also bind to human complement C1q. Thus, feline IgG1a and 1b are expected to induce strong effector function in vivo. The additional IgG detected does not bind to recombinant fFcγRI or fFcγRIII and has negligible binding to hC1q. Consequently, although this putative subclass is projected to have a similar serum half-life as the IgG1 alleles based on comparable in vitro affinity to FcRn, it may not elicit the effector responses mediated by fFcγRI or fFcγRIII. Further testing with native receptors and functional cell-based assays would confirm effector function capabilities of feline IgG subclasses; however this is the first report characterizing affinities of feline IgGs to their Fc receptors and helps pave the way for construction of feline-specific IgGs for therapeutic use.
authors.affiliation: Zoetis, Inc., Global Therapeutic Research, Veterinary Medicine Research and Development, 333 Portage St., Kalamazoo, MI 49007, USA. Electronic address: catherine.j.strietzel@zoetis.com.
authors.forename: Catherine J
authors.initials: CJ
authors.lastname: Strietzel
authors.affiliation: Zoetis, Inc., Global Therapeutic Research, Veterinary Medicine Research and Development, 333 Portage St., Kalamazoo, MI 49007, USA.
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Bergeron
authors.affiliation: Zoetis, Inc., Global Therapeutic Research, Veterinary Medicine Research and Development, 333 Portage St., Kalamazoo, MI 49007, USA.
authors.forename: Theodore
authors.initials: T
authors.lastname: Oliphant
authors.affiliation: Zoetis, Inc., Global Therapeutic Research, Veterinary Medicine Research and Development, 333 Portage St., Kalamazoo, MI 49007, USA.
authors.forename: Veronica T
authors.initials: VT
authors.lastname: Mutchler
authors.affiliation: Zoetis, Inc., Global Therapeutic Research, Veterinary Medicine Research and Development, 333 Portage St., Kalamazoo, MI 49007, USA.
authors.forename: Leszek J
authors.initials: LJ
authors.lastname: Choromanski
authors.affiliation: Zoetis, Inc., Global Therapeutic Research, Veterinary Medicine Research and Development, 333 Portage St., Kalamazoo, MI 49007, USA.
authors.forename: Graeme
authors.initials: G
authors.lastname: Bainbridge
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D002415:Cats / Q000235:genetics* / Q000276:immunology*; D003001:Cloning, Molecular; D015922:Complement C1q / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D006801:Humans; D007127:Immunoglobulin Constant Regions / Q000235:genetics / Q000378:metabolism; D007074:Immunoglobulin G / Q000145:classification / Q000235:genetics* / Q000378:metabolism*; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D011485:Protein Binding; D011961:Receptors, Fc / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D001613:beta 2-Microglobulin / Q000235:genetics / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D007127:Immunoglobulin Constant Regions; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D011961:Receptors, Fc; D001613:beta 2-Microglobulin; D015922:Complement C1q
keywords: Cat; Effector function; Fc receptor; Feline; IgG; Subclass