Grant-in-Aid for Overseas Scientific Survey.
|Research Institution||Osaka University Faculty of Dentistry|
HAMADA Shigeyuki Department of Oral Microbiology, Osaka University Faculty of Dentistry, 歯学部, 教授 (60028777)
KIYONO Hiros アラバマ大学, 歯学部, 教授
MCGHEE Jerry アラバマ大学, 医学センター, 教授
小川 知彦 大阪大学, 歯学部, 助教授 (80160761)
KIYONO Hiroshi Department of Oral Biology, University of Alabama at Birmingham
OGAWA Tomohiko Department of Oral Microbiology, Osaka University Faculty of Dentistry
MCGHEE Jerry R. Department of Microbiology, University of Alabama at Birmingham
|Project Fiscal Year
1989 – 1991
Completed(Fiscal Year 1991)
|Budget Amount *help
¥11,300,000 (Direct Cost : ¥11,300,000)
Fiscal Year 1991 : ¥4,000,000 (Direct Cost : ¥4,000,000)
Fiscal Year 1990 : ¥3,000,000 (Direct Cost : ¥3,000,000)
Fiscal Year 1989 : ¥4,300,000 (Direct Cost : ¥4,300,000)
|Keywords||Periodontal disease / Fimbrial protein / Immunoglobulin classes / Antibody-secreting cells / Immune response / Gingival tissues / Periodontopathic bacteria / Administration route / 歯周病 / 線毛タンパク / 免疫グロブリンクラス / 抗体産生細胞 / 免疫応答 / 歯肉組織 / 歯周病原性細菌 / 投与経路 / クラス|
Porphyromonas (Bacteroides) gingivalis, an anaerobic, black-pigmented rod, has been recognized as a major etiologic agent of periodontal disease. The organisms have peritrichous fimbriae ; the fimbriae were detached from fresh whole cells of P. gingivalis strain 381 by gentle pipetting and purified chromatographically using a DEAESepharose Fast Flow column. The purified fimbrial protein gave a single band of 41 KDa.
Oral administration of P. gingivalis fimbriae with adjuvant GM-53 in liposomes, but not in Tris-HCI buffer, definitely enhanced the fimbria-specific serum IgG responses (IGGI>>IgG2b>IgG2a>IgG3) and IgA response in saliva of BALB/C mice. On the other hand, subcutaneous (s. c.) injection of fimbriae with GM-53 also raised the fimbria-specific IgG followed by IgA and IgM responses in serum, and both IgA and IgG responses in saliva. Oral immunization was less effective than s. c. injection in terms of the production of serum antibody in the mice. However, the level of salivary a
ntibody of mice injected s. c. was similar to that of mice immunized orally. High anti-fimbriae IgG in serum were maintained in BALB/c mice immunized orally with fimbriae and GM-53 in liposomes for ca. 7 months after the primary immunizations. Oral administration also induced and held the fimbriaspecific IgA response in saliva for at least 6 months after the primary immunizations.
We have shown that patients with adult periodontitis (AP) exhibited elevated serum IgG antibody levels to P. gingivalis fimbriae. The subclass response was IgG3>>IgGI>IgG2>>IgG4. Some IgA anti-fimbriae antibodies were also observed. IgAl predominated, but significant levels of IgA2 were also seen.
We then assessed numbers of anti-fimbriae antibody-secreting cells from peripheral blood mononuclear cells (PBMC) and from inflamed gingiva of AP subjects by using a newly developed ELISPOT method. Gingival mononuclear cells (GMC) of AP subjects exhibited high numbers of spot-forming cells (SFC) including fimbriaespecific antibody secreting cells in a pattern of IgG>IgA>>IgM, while low numbers of SFC were seen in GMC from healthy gingiva. No fimbriae-specific SFC were seen in PBMC. It was also found GMC released increased levels of IL-5, IL-6 and TGF-beta but not IL-2 and IL4 when they were grown in vitro.
Thus, we have examined pathogenic mechanisms of P. gingivalis in the development of periodontal disease from the veiw points of microbiology and immunology. Less