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）已被用来调查潜在的贡献，宿主防御牙龈卟啉单胞菌感染。通过对存活细菌的测定和~ 3 H-胸腺嘧啶核苷掺入新合成的DNA的测定，我们发现这些抗体在豚鼠补体存在时对牙龈卟啉单胞菌具有显著的杀伤活性。此外，我们证实了在补体源存在下暴露于单克隆抗体的牙龈卟啉单胞菌381在放射性杀细菌试验中的细胞裂解，该试验使用了用13 <14>C-乙酸钠标记的细菌。这些数据表明，40 kDa的OMP将是有效的目标，以开发针对牙龈卟啉单胞菌的疫苗。接下来，我们从完整的DNA序列中分析了40-kDa OMP的氨基酸序列。平均筛选出6种多肽作为多肽组分疫苗，并合成了这些多肽（约20个残基）。抗40-kDa OMP的多克隆抗体识别合成的6个肽中的2个。合成多抗原肽（MAP），免疫家兔。抗血清能识别MAP抗原，但不能识别40-kDa OMP，因此我们将该肽与二氢叶酸还原酶偶联，再次免疫。抗血清能与40-kDa OMP发生反应，因此，合成的多肽可用于牙周病组分疫苗的研制。