Autoreactive sites of human lambda light chain mapped by comprehensive peptide synthesis.
Autoantibodies reactive against immunoglobulins are associated with autoimmune disorders as well as with immunization and infection. Moreover, recent interest is focused on auto-antidiotypes because of their possible role in immunoregulation. In this study, we used a set of overlapping synthetic peptides duplicating the structure of the monoclonal human lambda light chain Mcg to map autoreactive determinants recognized by natural antibodies present in normal polyclonal human IgG. We found that autoantibodies in human IgG react strongly with two distinct V lambda determinants corresponding to the first complementarity determining region (CDR1) and the third framework (Fr3). Antibodies showing weak reactivities against three regions of the constant domain also occur in the preparations. The antibodies directed against light chain peptides comprise less than 0.1% of the IgG pool. Analysis by direct binding and by competitive ELISA inhibition established that affinity purified antibodies specific for CDR1 and Fr3 peptide determinants react with the intact light chain Mcg as well as with the corresponding peptide. Competitive inhibition studies comparing total IgG and affinity-purified antibodies indicate that natural antibodies showing a wide range of affinities are present. The polyclonal nature of the natural antibodies is further shown by the presence of both kappa and lambda light chains in the purified antibodies. Although the role of such natural antibodies remains to be determined, the cross-reactivity between V lambda peptides and the intact chain suggest that they can function in regulation of antibody formation.
Kaymaz
H
H
Department of Microbiology and Immunology, College of Medicine, University of Arizona, Tucson 85724.
Marchalonis
J J
JJ
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000378:metabolism; D001323:Autoantibodies / Q000737:chemistry* / Q000378:metabolism; D001666:Binding Sites, Antibody; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry* / Q000378:metabolism; D007145:Immunoglobulin kappa-Chains / Q000737:chemistry* / Q000378:metabolism*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000138:chemical synthesis / Q000737:chemistry; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry*
D000911:Antibodies, Monoclonal; D001323:Autoantibodies; D007074:Immunoglobulin G; D007145:Immunoglobulin kappa-Chains; D009842:Oligopeptides; D010455:Peptides