Identification and characterization of B-cell epitopes of a 53-kDa outer membrane protein from Porphyromonas gingivalis.
We have previously reported that Porphyromonas gingivalis FDC 381 possesses a 53-kDa protein antigen (Ag53) on its outer membrane that evokes a strong humoral immune response in many patients with periodontal disease and that the humoral immune responses to Ag53 differ greatly among patients. To understand how the individual humoral immune system against Ag53 was determined, the regions of Ag53 recognized by specific antibody (B-cell epitopes) and dominant subclasses of serum immunoglobulin G (IgG) against major B-cell epitopes were examined by enzyme-linked immunosorbent assay. This study used sera from six patients with periodontitis, which all reacted strongly with sonic extracts of P. gingivalis 381 and with purified Ag53, and sera from six periodontally healthy children, which did not react with either sonic extracts of P. gingivalis 381 or Ag53. The epitopes were identified using synthetic 5-residue overlapping decapeptides covering the entire Ag53. Thirteen of 89 synthetic decapeptides showed a strong reaction with sera from the periodontal patients, but no reaction with those from the healthy children. Four peptides of 13 exerted different immune responses among patients. Furthermore, restriction analyses of the highly antigenic regions revealed that three sequences, RAAIRAS, YYLQ and MSPARR, were identified as major B-cell epitopes. Additionally, these epitopes were recognized mainly by the IgG2 isotype. These data suggest that the difference of B-cell epitopes might influence individual differences in antibody titer against Ag53 and also that the epitopes recognized commonly by multiple antibodies are quite valuable for peptide vaccine development against P. gingivalis infection.
Department of Periodontology and Endodontology, Okayama University Dental School, 2-5-1 Shikata-cho, Okayama 700-8525, Japan.
K
K
Oyaizu
H
H
Ohyama
F
F
Nishimura
H
H
Kurihara
S
S
Matsushita
H
H
Maeda
S
S
Kokeguchi
H
H
Hongyo
S
S
Takashiba
Y
Y
Murayama
D000293:Adolescent; D000328:Adult; D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000276:immunology; D016022:Case-Control Studies; D002648:Child; D002675:Child, Preschool; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000737:chemistry* / Q000276:immunology; D005260:Female; D006801:Humans; D016056:Immunodominant Epitopes / Q000737:chemistry / Q000276:immunology; D007074:Immunoglobulin G / Q000276:immunology; D008297:Male; D009842:Oligopeptides / Q000302:isolation & purification; D010518:Periodontitis / Q000097:blood / Q000276:immunology*; D016966:Porphyromonas gingivalis / Q000276:immunology*
C116911:Ag53 protein, Porphyromonas gingivalis; D001425:Bacterial Outer Membrane Proteins; D018985:Epitopes, B-Lymphocyte; D016056:Immunodominant Epitopes; D007074:Immunoglobulin G; D009842:Oligopeptides