Mapping of murine Th1 and Th2 helper T-cell epitopes on fimbriae from Porphyromonas gingivalis.
Th1- and Th2-derived cytokine production in response to synthetic peptides of the fimbrial subunit protein (fimbrilin) from Porphyromonas gingivalis strain 381 was assessed in spleen mononuclear cells (MNC) of BALB/c mice (H-2d haplotype) immunised with the fimbrial protein antigen and adjuvant GM-53 in Freund's incomplete adjuvant (FIA). Sixty-seven sequential overlapping 10-mer peptides covering the complete 337 amino-acids (AA) protein of P. gingivalis fimbrilin were synthesised. Stimulation of spleen MNC in vitro with these 10-mer peptides resulted in the production of murine interleukin-2 (IL-2), gamma-interferon (IFN-gamma), IL-4, IL-5 and IL-6. Peptides 13 (AA 61-70), 24 (AA 116-125), 31 (AA 151-160) and 64 (AA 316-325) markedly induced IL-2 production. In particular, peptide 24 (DPLKIKRVHA), which contained I-Ad, I-Ed and I-Ak binding motifs, was the most potent stimulator of IL-2, IFN-gamma, IL-4, IL-5 and IL-6 production. Spleen MNC from C3H/HeN mice (H-2k) followed by BALB/c mice (H-2d) immunised with peptide 24 were high responders to peptide 24 in terms of both IFN-gamma and IL-4 production, whereas A/J mice (H-2a) and C57BL/6 mice (H-2b) were very low responders, P. gingivalis fimbriae evoked higher delayed-type hypersensitivity (DTH) reactions in B10.D2 (H-2d) and B10.BR (H-2k) mice followed by C57BL/10 (B10, H-2b) and B10.A (H-2a) and in guinea-pigs immunised with the fimbriae and GM-53 in FIA.(ABSTRACT TRUNCATED AT 250 WORDS)
Ogawa
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T
Department of Oral Microbiology, Osaka University Faculty of Dentistry, Japan.
Uchida
H
H
Yasuda
K
K
D000595:Amino Acid Sequence; D000818:Animals; D000942:Antigens, Bacterial / Q000737:chemistry / Q000276:immunology; D001426:Bacterial Proteins / Q000737:chemistry / Q000276:immunology; D016207:Cytokines / Q000096:biosynthesis; D018604:Epitope Mapping; D000939:Epitopes / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D036781:Fimbriae Proteins; D010861:Fimbriae, Bacterial / Q000276:immunology*; D006168:Guinea Pigs; D006968:Hypersensitivity, Delayed; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D016966:Porphyromonas gingivalis / Q000276:immunology* / Q000648:ultrastructure; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology*
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D016207:Cytokines; D000939:Epitopes; C063055:fimbrillin; D036781:Fimbriae Proteins