Elucidation of linear epitopes of pertussis toxin using overlapping synthetic decapeptides: identification of a human B-cell determinant in the S1 subunit indicative of acute infections.
To identify relevant linear epitopes within the immunodominant ADP-ribosyl transferase (S1 subunit) of pertussis toxin (PT), its complete amino acid sequence was synthesized as consecutive, overlapping decapeptides on solid phase and probed for seroreactivity with pertussis specific human antisera in 'peptide scans'. Comparison of the resulting antigenic profiles revealed two distinct types of human antisera, though amino acids 140-200 could not be assessed as the corresponding peptides reacted non-specifically with the detection system. Human anti-pertussis sera predominantly recognized linear immunodominant epitopes located in three separated segments spanning amino acids 3-16, 21-30, and 211-222. Antisera originating from infants with acute B. pertussis infections (type I) identified determinants in all three segments, while type-II antisera from convalescent patients only recognized epitopes in the N-terminal regions. The binding of pertussis specific antisera--both type I and type II--to the holotoxin was inhibited by preincubation of antibodies with synthetic peptides corresponding to two linear determinants located at the N-terminus of S1:R 3-16 and R 21-30. However, competitive binding of antibodies to PT and to synthetic peptides equivalent to the third epitope (R 211-222) was only observed with type I antisera. Thus, the linear immunogenic determinant identified at the C-terminus of the A-protomer represents a human epitope which is apparently specific for antisera from pertussis patients with acute infections. The possible application of this determinant in serologic diagnosis will be a valuable tool to detect and distinguish acute Bordetella pertussis infections.
Raupach
B
B
Institut für Infektiologie, Westfälische Wilhelms-Universität Münster, Germany.
Schmidt
M A
MA
D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology*; D001886:Bordetella pertussis / Q000276:immunology* / Q000378:metabolism; D002648:Child; D002675:Child, Preschool; D000939:Epitopes / Q000276:immunology* / Q000378:metabolism; D005260:Female; D006801:Humans; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology; D037342:Pertussis Toxin; D011485:Protein Binding; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000276:immunology* / Q000378:metabolism; D013045:Species Specificity; D014614:Vaccines, Synthetic; D010566:Virulence Factors, Bordetella / Q000276:immunology* / Q000378:metabolism; D014917:Whooping Cough / Q000175:diagnosis / Q000276:immunology*
D000907:Antibodies, Bacterial; D000939:Epitopes; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic; D010566:Virulence Factors, Bordetella; C083388:pertussis toxin, S1 subunit; D037342:Pertussis Toxin