In vivo cleavage of immunoglobulin A1 by immunoglobulin A1 proteases from Prevotella and Capnocytophaga species.
Immunoglobulin A1 (IgA1) proteases secreted by oral Prevotella and Capnocytophaga species specifically cleave IgA1 at the same peptide bond in the hinge region, leaving intact monomeric Fab and Fc fragments. Assuming that Prevotella- and Capnocytophaga-induced Fab fragments of IgA1 expose a specific immunogenic neoepitope at the cleavage site, we established an enzyme-linked immunosorbent assay to measure human serum antibodies to this neoepitope as indirect evidence of in vivo activity of Prevotella and Capnocytophaga IgA1 proteases. The assay used a monoclonal antibody with specificity for the neoepitope, and the ability to block binding of the monoclonal antibody to the neoepitope was investigated. Absorption of sera with Prevotella melaninogenica-induced Fab fragments of IgA1 resulted in removal of antibodies blocking binding of the monoclonal antibody, whereas absorption with Fab fragments induced by bacterial IgA1 proteases of other cleavage specificities did not remove blocking antibodies. Consequently, we assume that the antibodies detected had been induced by a neoepitope an the Fab fragment of IgA1 exposed exclusively after cleavage with IgA1 proteases from Prevotella and Capnocytophaga, indicating in vivo activity of these IgA1 proteases. Evidence, though indirect, of in vivo activity of Prevotella and Capnocytophaga IgA1 proteases was present in 42 of 92 sera examined and in a significantly higher proportion of sera from adults with periodontal disease compared with control individuals. No correlation with disease was observed for the juvenile periodontitis groups.
Frandsen
E V
EV
Department of Oral Biology, Royal Dental College, University of Aarhus, Denmark.
Reinholdt
J
J
Kjeldsen
M
M
Kilian
M
M
D000293:Adolescent; D000328:Adult; D010520:Aggressive Periodontitis / Q000382:microbiology; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood / Q000276:immunology; D000911:Antibodies, Monoclonal; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D002206:Capnocytophaga / Q000201:enzymology* / Q000276:immunology; D017753:Ecosystem; D000939:Epitopes / Q000276:immunology; D006801:Humans; D007070:Immunoglobulin A / Q000276:immunology / Q000378:metabolism; D007140:Immunoglobulin Fab Fragments / Q000276:immunology / Q000378:metabolism; D007132:Immunoglobulin Isotypes; D051379:Mice; D008807:Mice, Inbred BALB C; D008875:Middle Aged; D010518:Periodontitis / Q000382:microbiology; D018720:Prevotella / Q000201:enzymology* / Q000276:immunology; D012697:Serine Endopeptidases / Q000378:metabolism*
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000939:Epitopes; D007070:Immunoglobulin A; D007140:Immunoglobulin Fab Fragments; D007132:Immunoglobulin Isotypes; D012697:Serine Endopeptidases; C021433:IgA-specific serine endopeptidase