Theiler's virus as a vector for foreign gene delivery into the CNS

泰勒病毒作为外源基因传递至中枢神经系统的载体

• 批准号: 09670675
• 负责人: OHARA Yoshiro
• 金额: $ 1.86万
• 依托单位: Kanazawa Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670675/
• 关键词: Theiler's virus; CNS; lymphotoxin gene; vector; macrophages

We reported that DA strain of Theiler's murine encephalomyelitis virus (TMEV) grows in macrophages where persists in the chronic stage if infection, therefore maintaining the genome. On the other hand, GDVII strain of TMEV does not grow there, and cannot maintain the genome (J Virol 71 : 729-733, 1997). Such a strain-specific, host cell-dependent virus growth was observed only in macrophages, but not in other types of cells (Microbiol Immunol, 43 : 885-892, 1999). Recombinant and mutant virus study also demonstrated that LィイD1*ィエD1 protein, which is out-of-frame with the polyprotein and translated from the alternative AUG within the L coding region, plays an important role for this phenomenon (J Virol 72 : 4950-4955, 1998). We finally inserted lymphotoxin (LT) gene into the L coding region of DA strain of TMEV and generated the recombinant virus. Western blotting and enzyme-linked immunosorbent assay demonstrated the expression of LT in BHK-21 cells infected with the recombinant virus. In addition, the expressed LT showed cytotoxicity against L-929 cells. Therefore, the present study provides a beginning of the use of TMEV as a vector for foreign gene delivery into the CNS.

我们报道了泰勒氏小鼠脑脊髓炎病毒(TMEV)DA株在巨噬细胞中生长，如果感染，巨噬细胞将持续处于慢性期，从而维持基因组。另一方面，TMEV的GDVII毒株不在那里生长，不能维持基因组(J Virol 71：729-733,1997)。这种依赖宿主细胞的菌株特异性病毒只在巨噬细胞中观察到，而在其他类型的细胞中没有观察到(微生物免疫，43：885-892,1999)。重组和突变病毒研究也表明，LィイD1*ィエD1蛋白与多聚蛋白在框外，从L编码区内的替代AUG翻译而来，在此现象中起着重要作用(J Virol 72：4950-4955,1998)。将淋巴毒素(LT)基因插入TMEV DA株L编码区，构建成重组病毒。Western blotting和酶联免疫吸附试验证实重组病毒感染的BHK-21细胞表达LT。此外，表达的LT对L-929细胞具有细胞毒作用。因此，本研究为利用TMEV作为外源基因进入中枢神经系统的载体提供了一个开端。

项目成果

大原義朗: "Theilerウイルスー持続感染と脱髄の発症機序に関する一考察ー"ウイルス. 49. 175-181 (1999)

Yoshiro Ohara：“泰勒病毒 - 持续感染和脱髓鞘发病机制的研究”病毒。 49. 175-181 (1999)

Obuchi M et al: "Expression of lymphotoxin gene inserted into Theiler's murine encephalomyelitisi virus"Microbiol.Immunol.. 43. 83-86 (1999)

Obuchi M等人：“插入泰勒氏鼠脑脊髓炎病毒的淋巴毒素基因的表达”Microbiol.Immunol.. 43. 83-86 (1999)

Obuchi M et al: "TMEV and its persistence" Neuropathology. 18. 13-18 (1998)

Obuchi M 等人：“TMEV 及其持久性”神经病理学。

大原義朗: "神経感染症の疾患治療薬の分類と臨床薬里" 今月の治療. 6. 83-87 (1998)

Yoshiro Ohara：“神经感染性疾病药物的分类和临床药学”本月的治疗。 6. 83-87 (1998)。

大原義朗: "ウイルス感染後アレルギー性神経疾患" 日本臨床. 55. 800-804 (1997)

Yoshiro Ohara：“病毒感染后的过敏性神经系统疾病”日本临床 55. 800-804 (1997)