Development of avian component vaccine with built-in adjuvanting potencial.

开发具有内在佐剂潜力的禽类成分疫苗。

• 批准号: 07556119
• 负责人: KON Yasuhiro
• 金额: $ 3.2万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07556119/
• 关键词: recombinant vaccine; chicken; adjuvant; Marek's disease virus; Botulinum toxin; prokaryotic expression vector; Newcastle disease virus; avian influenza virus; 高速液体クロマトグラフィー; コンポーネントワクチン; ニューカッスル病; B抗原; 細胞接着

The aim of this project is the development of a component vaccine which have adjuvanting potential in itself. The strategy for adding adjuvanting property antigens is the use of recombinant protein translated from artificially joined construct of genes. We focused the adhesive property of the heavy chain of Botulinum toxin. Although the adhesion ability indicated to be located on the C-terminal half of the heavy chain, whole sized heavy chain was tested for adhesion ability. Antigen for component vaccine was selected from avian influenza, Newcastle disease virus, Turkey rhinotrachitis and Marek's disease virus. Additionally, we determined MDV gB as the first triat for adjuvanting ability. To obtain recombinant fusion protein (Btx-gB), we construced the gene fragment and ligated with E.coli-expression vector pET32. Antibody production was induced by injections of recombinant proteins into experimental chickens. While, the concentration of anti-gB antibody after injection of recombinant gB was not lower than that of anti-gB andtibody after injection of recombinant-fusion protein Btx-gB.This data indiceated that the Btx-gB fusion protein did not have enough adjuvanting property. Anothr construction should be tested.

该项目的目的是开发一种本身具有佐剂潜力的组分疫苗。添加佐剂特性抗原的策略是使用从人工连接的基因构建体翻译的重组蛋白。我们重点研究了肉毒毒素重链的粘附特性。虽然粘附能力表明位于重链的C-末端一半，但测试了整个大小的重链的粘附能力。从禽流感病毒、纽卡斯尔病病毒、火鸡鼻气管炎病毒和马立克氏病病毒中筛选出疫苗组分抗原。此外，我们确定MDV gB作为佐剂能力的第一个试验。构建Btx-gB基因片段，连接到大肠杆菌表达载体pET 32，获得重组融合蛋白（Btx-gB）。通过将重组蛋白注射到实验鸡中诱导抗体产生。而重组gB注射后的抗体浓度并不低于重组融合蛋白Btx-gB注射后的抗体浓度，说明Btx-gB融合蛋白没有足够的佐剂性。应测试另一个结构。

项目成果

Ito.T.: "Receptor specificity of influenza A viruses correlates with the aggliutination of erythrocytes from different animal species." Virology. 227. 493-499 (1997)

Ito.T.：“甲型流感病毒的受体特异性与不同动物物种的红细胞的凝集相关。”

Endoh, D.: "Expression of the endogeneous Marek's disease virus ICP4 homolog (MDV ICP4) gene is enhanced in latently infected cells by transient transfection with the recombinant MDV ICP4 gene." Japanese Journal of Veterinary Research. 43. 109-124 (1995)

Endoh, D.：“通过重组 MDV ICP4 基因瞬时转染，潜伏感染细胞中内源性马立克氏病病毒 ICP4 同源物 (MDV ICP4) 基因的表达得到增强。”

Endoh, D.: "Retroviral sequence inserted intoMd5 strain of Marek's disease virus type1." J.Vet.Med.Sci.60. 227-235 (1998)

Endoh, D.：“将逆转录病毒序列插入马立克氏病病毒 1 型 Md5 株中。”

Kon,Y.: "Expression of renin in the rat liver." Anatomia Histologia Embryologia. 27(in press). (1998)

Kon，Y.：“大鼠肝脏中肾素的表达。”

Kon, Y.: "Detection of coagulating gland renin by hybridohistochemistry." Anatomia Histologia Embryologia. 25. 289-294 (1996)

Kon，Y.：“通过杂交组织化学检测凝固腺肾素。”