title: Purification and characterisation of immunoglobulins from the Australian black flying fox (Pteropus alecto) using anti-fab affinity chromatography reveals the low abundance of IgA.
abstract: There is now an overwhelming body of evidence that implicates bats in the dissemination of a long list of emerging and re-emerging viral agents, often causing illnesses or death in both animals and humans. Despite this, there is a paucity of information regarding the immunological mechanisms by which bats coexist with highly pathogenic viruses. Immunoglobulins are major components of the adaptive immune system. Early studies found bats may have quantitatively lower antibody responses to model antigens compared to conventional laboratory animals. To further understand the antibody response of bats, the present study purified and characterised the major immunoglobulin classes from healthy black flying foxes, Pteropus alecto. We employed a novel strategy, where IgG was initially purified and used to generate anti-Fab specific antibodies. Immobilised anti-Fab specific antibodies were then used to capture other immunoglobulins from IgG depleted serum. While high quantities of IgM were successfully isolated from serum, IgA was not. Only trace quantities of IgA were detected in the serum by mass spectrometry. Immobilised ligands specific to IgA (Jacalin, Peptide M and staphylococcal superantigen-like protein) also failed to capture P. alecto IgA from serum. IgM was the second most abundant serum antibody after IgG. A survey of mucosal secretions found IgG was the dominant antibody class rather than IgA. Our study demonstrates healthy P. alecto bats have markedly less serum IgA than expected. Higher quantities of IgG in mucosal secretions may be compensation for this low abundance or lack of IgA. Knowledge and reagents developed within this study can be used in the future to examine class-specific antibody response within this important viral host.
authors: Australian Animal Health Laboratory, CSIRO Animal, Food and Health Sciences, Geelong, Victoria, Australia.
authors: James W
authors: JW
authors: Wynne
authors: Antonio
authors: A
authors: Di Rubbo
authors: Brian J
authors: BJ
authors: Shiell
authors: Gary
authors: G
authors: Beddome
authors: Christopher
authors: C
authors: Cowled
authors: Grantley R
authors: GR
authors: Peck
authors: Jing
authors: J
authors: Huang
authors: Samantha L
authors: SL
authors: Grimley
authors: Michelle L
authors: ML
authors: Baker
authors: Wojtek P
authors: WP
authors: Michalski
mesh_terms: D000818:Animals; D002685:Chiroptera / Q000276:immunology*; D002846:Chromatography, Affinity / Q000379:methods*; D007070:Immunoglobulin A / Q000032:analysis* / Q000302:isolation & purification; D007136:Immunoglobulins / Q000032:analysis* / Q000302:isolation & purification
chemical_list: D007070:Immunoglobulin A; D007136:Immunoglobulins