Antigenic and immunological mimicry of peptide mimotopes of Lewis carbohydrate antigens.
Peptides may substitute for carbohydrates in reactions with carbohydrate-specific molecules. Recently, we found that peptides containing aromatic residues mimic mucin and histo-blood group related carbohydrate epitopes, eliciting polyclonal responses cross-reactive with bacterial and viral antigens that express these carbohydrate forms. These results demonstrate that peptides can function in in vivo and in vitro models as carbohydrate surrogate antigens. To further explore the nature of the antigenic and immunogenic properties of such mimotopes, synthetic peptides with aromatic amino acids were tested to delineate reactivity patterns with several anti-neolactoseries monoclonal antibodies (MAbs). These MAbs recognize biologically important conformations of the histo-blood group related Lewis antigens expressed on the surface of a variety of human cancers. Results by ELISA demonstrate that the MAbs can distinguish particular peptide motifs that include the sequences GGIYYPYDIYYPYDIYYPYD, GGIYWRYDIYWRYDIYWRYD and GGIYYRYDIYYRYDIYYRYD. Substitution of Arg by Pro diminished the reactivity of the anti-Lewis Y (LeY) MAb BR55-2. Binding of LeY to BR55-2 was inhibitable by the Arg containing peptides. Serum against all three peptides displayed reactivity with synthetic histo-blood group related antigen probes. Immunologic presentation of the peptides as multiple antigen peptides (MAPs) improved peptide ability to induce LeY specific immune responses. Serum bound to human tumor cells that preferentially expressed neolactoseries antigens, but not to normal tissues. Immunoprecipitation of human breast tumor cell lysates before and after treatment with tunicamycin confirmed serum carbohydrate binding. The anti-peptide sera mediated tumor cell killing by complement mediated cytotoxicity. These results indicate that mapping peptide epitopes with anti-carbohydrate antibodies can lend to defining antibody fine specificities that can go undetected by screening of carbohydrate antigens alone. In addition, these results confirm that peptides and carbohydrates can bind to the same antibody binding site and that peptides can structurally mimic salient features of carbohydrate epitopes.
Luo
P
P
Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
Agadjanyan
M
M
Qiu
J
J
Westerink
M A
MA
Steplewski
Z
Z
Kieber-Emmons
T
T
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000737:chemistry / Q000276:immunology; D000939:Epitopes / Q000235:genetics; D006801:Humans; D007983:Lewis Blood Group Antigens / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D018716:Molecular Mimicry / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D010455:Peptides / Q000235:genetics / Q000276:immunology; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D000939:Epitopes; D007983:Lewis Blood Group Antigens; C094162:Lewis Y antigen; D010455:Peptides