IgM natural autoantibodies against bromelain-treated mouse red blood cells recognise carbonic anhydrase.
Carbonic anhydrase (CA) from mouse erythrocyte membranes is recognised as an autoantigen in Western blotting experiments with FUB 1, a murine IgM monoclonal antibody that binds both phosphatidylcholine and bromelain-treated mouse red blood cells (BrMRBC). Serum from mice stimulated with lipopolysaccharide (LPS-serum) also recognises CA. From SDS-PAGE, and blotting experiments with whole mouse erythrocytes, we found two closely spaced glycoprotein bands in the 30 kD region that reacted with both FUB 1 and LPS-serum. One of the molecular weight markers, bovine carbonic anhydrase which is of a molecular weight of about 30 kD, electrophoresed in the same 30 kD region also reacted with these antibodies. Carbonic anhydrases from a range of mammalian species were found to be crossreactive with FUB 1 and LPS-serum by Western blotting, whereas human glycophorin A and human asialoglycophorin were not recognised by the antibodies. FUB 1 specifically recognises both native and denatured bovine carbonic anhydrase in ELISA assays. The serological identity of the determinants of CA and BrMRBC was confirmed by specific absorption of both FUB 1 and LPS-serum with BrMRBC and normal mouse erythrocytes. We propose that a native autoantigenic epitope on erythrocytes may be revealed by the proteolytic action of bromelain and that this determinant is associated, at least in part, with carbonic anhydrase.
Jonusys
A M
AM
Department of Biology, University of Wollongong, NSW, Australia.
Cox
K O
KO
Steele
E J
EJ
D000818:Animals; D001323:Autoantibodies / Q000276:immunology*; D001667:Binding, Competitive; D001963:Bromelains / Q000494:pharmacology; D002256:Carbonic Anhydrases / Q000737:chemistry / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D004912:Erythrocytes / Q000187:drug effects / Q000276:immunology*; D005260:Female; D015151:Immunoblotting; D007075:Immunoglobulin M / Q000276:immunology*; D051379:Mice; D008815:Mice, Inbred Strains
D001323:Autoantibodies; D000939:Epitopes; D007075:Immunoglobulin M; D001963:Bromelains; D002256:Carbonic Anhydrases