| Project/Area Number |
12670263
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Bacteriology (including Mycology)
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| Research Institution | Chukyo Women's University |
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Principal Investigator |
KOZUKA Satoshi Faculty of Wellness, Associate Professor, 健康科学部, 助教授 (40117817)
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| Co-Investigator(Kenkyū-buntansha) |
NOBUSAWA Eri Nagoya City University, School of Medicine, Associate Professor, 医学部, 助教授 (90183904)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 2001: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2000: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | pertussis / wooping cough / recombinant vaccine / substituted PTA / intranasal administration / Bacillus brevis / rCTB / rLTB / 百日咳ワクチン / 経鼻接種 / 粘膜ワクチン / アジュバント / CTB / LTB / 無毒化PTA |
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| Research Abstract |
A gene encoding the amino acid substituted Bordetella pertussis pertussis toxin A. subunit (sPTA-9K/129G ; Arg^9→Lys and Glu^<129>→Gly) was introduced in a vector pNU212 and expressed at high levels in Bacillus brevis. The colony obtained showed a positive reaction to anti-PT antibody in an in situ colony immunoassay of B. brevis. A transformant producing sPTA on the largest scale was chosen and used as B. brevis HPD31 carring pNU212-sPTA in this study. When B. brevis carring pNU212-sPTA was cultivated with shaking at 30 C in modified 5PY medium, sPTA reached a maximum after 5 days' cultivation. The maximum amount of sPTA secreted into medium was 50 mg /L when determined from the depth of color on the protein bands of SDS-PAGE. Such strain is therefore suitable for large-scale production of an intranasal acellular wooping cough vaccine by genetically detoxified PTA with mucosal adjuvant.
Furthermore, we have shown that recombinant CTB (rCTB) produced by B. brevis carring pNU212-CTB or recombinant LTB (rLTB) produced by it carring pNU212-LTB have an adjuvant activity when administerd intranasally with alminium-non-adsorbed diphtheria toxoid. It resulted in the substantial stimulation of DT-specific serum antibody, and in the induction of DT-specific mucosal IgA antibody responses in the nasal cabity, the lung, the saliva and vaginal secretion. These results show that intranasal administration of PT with rCTB or rLTB must be a very useful means for vaccination against pertussis.
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