Human anti-IgG1 hinge autoantibodies reconstitute the effector functions of proteolytically inactivated IgGs.
A number of proteases of potential importance to human physiology possess the ability to selectively degrade and inactivate Igs. Proteolytic cleavage within and near the hinge domain of human IgG1 yielded products including Fab and F(ab')(2) possessing full Ag binding capability but absent several functions needed for immune destruction of cellular pathogens. In parallel experiments, we showed that the same proteolytically generated Fabs and F(ab')(2)s become self-Ags that were widely recognized by autoantibodies in the human population. Binding analyses using various Fab and F(ab')(2), as well as single-chain peptide analogues, indicated that the autoantibodies targeted the newly exposed sequences where proteases cleave the hinge. The point of cleavage may be less of a determinant for autoantibody binding than the exposure of an otherwise cryptic stretch of hinge sequence. It was noted that the autoantibodies possessed an unusually high proportion of the IgG3 isotype in contrast to Abs induced against foreign immunogens in the same human subjects. In light of the recognized potency of IgG3 effector mechanisms, we adopted a functional approach to determine whether human anti-hinge (HAH) autoantibodies could reconstitute the (missing) Fc region effector functions to Fab and F(ab')(2). Indeed, in in vitro cellular assays, purified HAH autoantibodies restored effector functions to F(ab')(2) in both Ab-dependent cellular cytotoxicity and complement-dependent cytotoxicity assays. The results indicate that HAH autoantibodies selectively bind to proteolytically cleaved IgGs and can thereby provide a surrogate Fc domain to reconstitute cell lytic functions.
Brezski
Randall J
RJ
Discovery Research, Centocor R&D Inc., Radnor, PA 19087, USA. rbrezski@cntus.jnj.com
Luongo
Jennifer L
JL
Petrone
Diane
D
Ryan
Mary H
MH
Zhong
Degang
D
Tam
Susan H
SH
Schmidt
Albert P
AP
Kruszynski
Marian
M
Whitaker
Brian P
BP
Knight
David M
DM
Jordan
Robert E
RE
D000936:Antigen-Antibody Complex; D001323:Autoantibodies / Q000276:immunology* / Q000378:metabolism; D001324:Autoantigens; D001666:Binding Sites, Antibody; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology* / Q000378:metabolism; D007074:Immunoglobulin G / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000378:metabolism*
D000936:Antigen-Antibody Complex; D001323:Autoantibodies; D001324:Autoantigens; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D010447:Peptide Hydrolases