Basic research for the development of mucosal vaccines using cholera toxin B subunit as an adjuvant

以霍乱毒素B亚基为佐剂的粘膜疫苗研制的基础研究

• 批准号: 10670266
• 负责人: TOCHIKUBO Kunio
• 金额: $ 2.11万
• 依托单位: Nagoya City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670266/
• 关键词: Recombinant CTB; Mucosal adjuvant; Intranasal administration; Mucosal vaccine; Tetanus toxoid; Diphtheria toxoid; Hepatitis B vaccine; CFA; III; 線維状赤血球凝集素; 志賀毒素1,2Bサブユニット; 感染防御

For the purpose of changing the immunization procedure of several vaccines from subcutaneous injection, which have been in practice for a long time, to intranasal administration, we have been examining systemic and mucosal immune responses of mice to some vaccines inoculated intranasally with recombinant cholera toxin B subunit (rCTB). Intranasal co-administration of rCTB with tetanus toxoid (TT), diphtheria toxoid (DT), pertussis component vaccine or hepatitis B vaccine (HBV) induced high levels of vaccine-specific serum IgG and mucosal IgA antibody titers. Moreover, a combination of rCTB with TT and DT provided complete protection against tetanus toxin challenge and diphtheria antitoxin titers above a protective level of 0.1 IU/ml, respectively. Intranasal immunization of HBV with rCTB also provided serum anti-HBV antibody titers above 100 mIU/ml protective in people at increased risk or displaying risk behavior. Pilus colonization factor antigen (CFA/III) of human enterotoxigenic Es … More cherichia coli (ETEC) was incorporated into the nasal mucosal epithelium without rCTB and induced CFA/ III-specific serum IgG and mucosal IgA antibody, suggesting the possibility of intranasal vaccine for preventing the disease produced by the ETEC.When five intranasal immunizations of rCTB and TT and subsequent five intranasal administrations with rCTB plus DT were performed, anti-rCTB antibodies induced by the previous vaccination did not have any effects upon the production of anti-DT antibodies, especially, at long intervals between two kinds of inoculations. Recombinant CTB elicited no toxic effects to macrophages and no vascular permeability-increasing effects. Moreover, no distinct local histopathological reactions were observed in the nasal cavity, the small-intestinal loop or the muscle given rCTB.These facts indicate that rCTB is a safe and useful candidate as a mucosal vaccine adjuvant.We finished constructing an efficient synthesis and secretion system for Shiga toxin I and II B subunits of enterohemorrhagic E.coli (EHEC) usins Bacillus brevis and have been purifing them to develop intranasal or oral vaccine for preventing hemorrhagic colitis and hemolytic-uremic syndrome produced by EHEC. Less

为了将长期以来使用的几种疫苗的免疫程序从皮下注射改为鼻内给药，我们研究了经鼻内接种重组霍乱毒素B亚单位（rCT B）的几种疫苗对小鼠的全身和粘膜免疫应答。rCTB与破伤风类毒素（TT）、白喉类毒素（DT）、百日咳组分疫苗或乙型肝炎疫苗（HBV）鼻内共施用诱导高水平的疫苗特异性血清IgG和粘膜IgA抗体滴度。此外，rCTB与TT和DT的组合分别提供了针对破伤风毒素攻击的完全保护和高于0.1 IU/ml的保护水平的白喉抗毒素滴度。用rCTB鼻内免疫HBV还提供了高于100 mIU/ml的血清抗HBV抗体滴度，在风险增加或表现出风险行为的人群中具有保护作用。人肠毒素E的菌毛定植因子抗原（CFA/III） ...更多信息 将大肠埃希菌（ETEC）掺入鼻粘膜上皮中，诱导产生CFA/III特异性血清IgG和粘膜IgA抗体，提示鼻内疫苗预防ETEC引起的疾病的可能性。由先前接种诱导的抗rCTB抗体对抗DT抗体的产生没有任何影响，尤其是在两种接种之间的长间隔下。重组CTB对巨噬细胞无毒性作用，也无血管通透性增加作用。此外，在鼻腔中未观察到明显的局部组织病理学反应，本研究成功构建了肠出血性大肠杆菌（EHEC）滋贺毒素I和II B亚单位的高效合成和分泌系统，并在此基础上构建了重组人志贺毒素B（rCTB）的分泌系统，为进一步研究肠出血性大肠杆菌（EHEC）志贺毒素B的分泌机制奠定了基础。利用短芽孢杆菌，并对其进行了纯化，以开发用于预防EHEC引起的出血性结肠炎和溶血性尿毒症综合征的鼻内或口服疫苗。少

项目成果

Isaka M, Yasuda Y, Kozuka S, Taniguchi T, Miura Y et al.: "Intranasal or subcutaneous co-administration of recombinant cholera toxin B subunit stimulates only a slight or no level of the specific IgE response in mice to tetanus toxoid" Vaccine. Vol.17, No

Isaka M、Yasuda Y、Kozuka S、Taniguchi T、Miura Y 等人：“重组霍乱毒素 B 亚基的鼻内或皮下联合给药仅刺激小鼠对破伤风类毒素的特异性 IgE 反应水平轻微或无水平”疫苗

Isaka M, Yasuda Y, Mizokami M et al.: "Mucosal immunization against hepatitis B virus by intranasal co-administration of recombinant hepatitis B surface antigen and recombinant cholera toxin B subunit as an adjuvant"Vaccine. 19 (11/12). 1460-1466 (2001)

Isaka M、Yasuda Y、Mizokami M 等人：“通过鼻内共同施用重组乙型肝炎表面抗原和重组霍乱毒素 B 亚基作为佐剂来对抗乙型肝炎病毒的粘膜免疫”疫苗。

Taniguchi T,Yasuda Y,Tochikubo T,Yamamoto K and Honda T: "The gene encoding the prepilin peptidase involved biosynthesis of pilus colonization factor antigen III(CFA/III) of human enterotoxigenic Escherichia coli"Microbiology and Immunology. Vol.43,No.9.

Taniguchi T、Yasuda Y、Tochikubo T、Yamamoto K 和 Honda T：“编码前菌毛蛋白肽酶的基因涉及人产肠毒素大肠杆菌的菌毛定植因子抗原 III (CFA/III) 的生物合成”微生物学和免疫学。

Tochikubo K,Isaka M,Yasuda Y,Kozuka S,Matano K et al.: "Recombinant cholera toxin B subunit acts as an adjuvant for the mucosal and systemic responses of mice to mucosally co-administered bovine serum albumin"Vaccine. Vol.16,No.2/3. 150-155 (1998)

Tochikubo K、Isaka M、Yasuda Y、Kozuka S、Matano K 等人：“重组霍乱毒素 B 亚基作为小鼠对粘膜共同施用牛血清白蛋白的粘膜和全身反应的佐剂”疫苗。

Isaka M,Yasuda Y,Kozuka S,Taniguchi T,Miura Y et al.: "Intranasal or subcutaneous co-administration of recombinant cholera toxin B subunit stimulates only a slight or no level of the specific IgE response in mice to tetanus toxoid"Vaccine. Vol.17,No.7/8.

Isaka M、Yasuda Y、Kozuka S、Taniguchi T、Miura Y 等人：“重组霍乱毒素 B 亚基的鼻内或皮下联合给药仅刺激小鼠对破伤风类毒素的特异性 IgE 反应水平轻微或无水平”疫苗