Development of Immunogene Therapy for B-cell malignancy

B细胞恶性肿瘤免疫基因疗法的发展

• 批准号: 09557083
• 负责人: SAITO Hidehiko
• 金额: $ 7.55万
• 依托单位: NAGOYA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09557083/
• 关键词: Idiotype Vaccine; Leukemia; Immunogene; Idiotype; idiotype; ワクチン; CTL; NAGL-1; マテリアル; 薬剤の安全性

(1) Material Production and Safety IsuueWe have tested followeing examination on our product : Abnormal Toxicity, LAL Test for Endotoxin, Detection of Contaminating DNA, Rabbit Pyrogen Test, Sterility Assay. No abnoramal data was observed on our materials.(2) Developement of New Vaccine Vector.We have constructed a novel DNA expression vector based on Semliki Forest virus (SFV) . A recombinant cDNA genome of SFV, in which the SFV structural genes were replaced by a polylinker cassette to allow for insertion of heterologous DNA, was placed under the control of a cytomegalovirus immediate-early enhancer/promoter with a polyadenylation signal. Transfection of mammalian cells with this SFV-based plasmid vector, pSFV3-CMV-lacZ-pA, resulted in transient high-level expression of a beta-galactosidase reporter gene.(3) Preparation for Clinical Trail.We have collected over hundred smples of leukemia patients for the vaccine production. Protocol for idiotype vaccination including Informed Consent and basic data was prepared for publish.

(1)材料生产和安全问题我们对产品进行了以下检查：异常毒性，内毒素LAL试验，DNA污染检测，家兔热原试验，无菌试验。在我们的材料上没有观察到异常数据。(2)新型疫苗载体的构建我们构建了一种基于Semliki森林病毒（SFV）的新型DNA表达载体。SFV的重组cDNA基因组，其中SFV结构基因被多接头盒取代以允许插入异源DNA，被置于具有多腺苷酸化信号的巨细胞病毒立即早期增强子/启动子的控制下。用这种基于SFV的质粒载体pSFV 3-CMV-lacZ-pA转染哺乳动物细胞，导致β-半乳糖苷酶报告基因的瞬时高水平表达。(3)临床试验的准备：我们已经收集了上百例白血病患者的样本用于疫苗的生产。独特型疫苗接种方案，包括知情同意书和基本数据，准备出版。

项目成果

AKIHIRO ABE et al: "Enhanced Gene Transfer with Fusogenic Liposomes Containing Vesicular Stomatitis Virus G Glycoprotein" JOURNAL OF VIROLOGY. 72(7). 6159-6163 (1998)

AKIHIRO ABE 等人：“使用含有水泡性口炎病毒 G 糖蛋白的融合脂质体增强基因转移”病毒学杂志。

Nobuhiko Emi: "Gene Transfer Mediated by Polyarginine Requires a Formation ofnBig Carrier-Complex of DNA Aggregate." Biochem. Biophys. Res. Commn.231. 421-424 (1997)

Nobuhiko Emi：“聚精氨酸介导的基因转移需要形成 DNA 聚集体的大载体复合物。”

KOHNO Akio: "Semliki Forest virus-based DNA expression vector : transient protein production followed by cell death" Gene Therapy. 5. 415-418 (1998)

KOHNO Akio：“基于 Semliki 森林病毒的 DNA 表达载体：瞬时蛋白质产生，随后细胞死亡”基因治疗。

AKIHIRO ABE: "In Vitro Cell-Free Conversion of Noninfectious Moloney Retrovirus Particles to an Infectious Form by the Addition of the Vesicular Stomatitis Virus Surrogate Envelope G Protein" JOURNAL OF VIROLOGY. 72(8). 6356-6361 (1998)

AKIHIRO ABE：“通过添加水泡性口炎病毒替代包膜 G 蛋白，将非感染性莫洛尼逆转录病毒颗粒体外无细胞转化为感染性形式”病毒学杂志。

KOHNO Akio et al: "Semliki Forest virus-based DNA expression vector : transient protein production followed by cell death." Gene Therapy. 5. 415-418 (1998)

KOHNO Akio 等人：“基于 Semliki 森林病毒的 DNA 表达载体：瞬时蛋白质产生，随后细胞死亡。”