Development of peptide component vaccine against P.gingivalis

牙龈卟啉单胞菌肽成分疫苗的研制

• 批准号: 06671871
• 负责人: ABIKO Yoshimitsu
• 金额: $ 0.45万
• 依托单位: NIHON UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06671871/
• 关键词: Porphyromonas gingivalis; bactericidal activity; monoclonal antibody; recombinant outer membtane protein; periodintitis; リコンビナ-ト外膜タンパク質; P.gingivalis; ペプチドワクチン; 合成ペプチド

Polyclonal antibody and monoclonal antibodies (MAb) directed against a recombinant 40-kDa outer membrane protein (OMP) have been used to investigate the potential contribution of host defense against infection of P.gingivalis. By the assay for the surviving bacteria and the incorporation of 3H-thymidine into newly synthesized DNA,we found significant killing activity for P.gingivalis by these antibodies when guinea pig complement was present. Additionally, we confirmed the cell lysis of P.gingivalis 381 exposed to the MAb in the presence of complement sources in a radioactive bacericidal assay using bacteria labelled with ^<14>C-sodium acetate. These data suggest that 40kDa OMP will be effective target to develop the vaccine against P.gingivalis. Next we analyzed amino acid sequences of 40-kDa OMP from complete DNA sequences. 6 peptides were selected by mean as peptide component vaccine and synthesized these peptides (about 20 reidues). Polyclonal antibody against 40-kDa OMP recognized 2 among 6 peptides synthesized. Then we synthesized multi-antigen peptides (MAP) and immunized to rabbits. Antisera recogniezed the MAP antigen, however did not recognized 40-kDa OMP.Therefore, we conjugated the peptide to dihydrofolate reductase and immunized again. The antiserem reacted to 40-kDa OMP.In conclusion therefore, synthesized peptide will be usefull to develop the component vaccine against periodontal disease.

针对重组40 kDa外膜蛋白（OMP）的多克隆抗体和单克隆抗体（MAB）已用于研究宿主防御抗抑制感染的潜在贡献。通过对幸存细菌的测定，并将3H-胸苷掺入新合成的DNA中，当存在豚鼠补体时，我们发现这些抗体对这些抗体的杀伤活性显着。此外，我们确认使用乙酸 ^<14> c-硫酸盐标记的细菌在放射性大量分析中存在补体源，暴露于mab的细胞裂解。这些数据表明，40KDA OMP将是开发针对吉利亚病的疫苗的有效靶标。接下来，我们分析了从完整的DNA序列中40 kDa OP的氨基酸序列。通过平均为肽成分疫苗选择6个肽，并合成这些肽（约20个再生）。对40 kDa OMP的多克隆抗体在合成的6种肽中识别2。然后，我们合成了多抗原肽（MAP）并将其免疫到兔子中。安提塞拉（Antisera）识别地图抗原，但是没有识别出40 kDa的能力。因此，抗血清对40 kDa的反应，因此，合成的肽对于开发针对牙周疾病的成分疫苗将是有用的。