Attempt for gene therapy of retrovirus-induced disease

逆转录病毒引起的疾病的基因治疗尝试

• 批准号: 09670219
• 负责人: KITAGAWA Masanobu
• 金额: $ 1.15万
• 依托单位: Tokyo Medical and Dental 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 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670219/
• 关键词: Retrovirus; Friend leukemia virus; Fv-4; Gene therapy

To develop the model for gene therapy of retrovirus-induced disease, Fv-4 resistance (Fv-4^r) gene which confers strong resistance against murine leukemia virus (MuLV)-infection to host has been introduced to Friend leukemia virus (FLV)-susceptible C3H mouse bone marrow cells.1) Bone marrow transplantation of Fv-4^r-transfected bone marrow cells to C3H hostAfter introducing Fv-4^r gene using SFFV vector system, C3H bone marrow cells were transplanted to supralethally irradiated C3H host to create bone marrow chimeras (Fv-4^r-G3H * C3H). Fv-4^r gene product was expressed on the cell surface of hematopoietic cells in these chimera mice. The intensity of expression varied from mouse to mouse most of which expressed ranging from 10% to 20% of the expression by the hematopoietic cells of genetically Fv-4^r-bearing C4W mouse.2) Development of gene induction system with high efficiencyBy selecting vector systems and culture conditions, we could generate Fv-4^r-C3H * C3H chimeras that exhibited Fv-4^r expression of more than 30% of C4W mouse.3) Trial experiment to test the resistance of these chimeras to FLV-infectionA few number of Fv-4^r-C3H * C3H chimeras with high expression of Fv-4^r (>30% of C4W) exhibited resistance against FLV inoculation of 1x103 PFU.However, all mice died after infection with lx 104 PFU of FLV.By using Fr-4^r-G3H * C3H chimeras with high expression of Fv-4^r which were successfully conducted in the latest experiments, we would be able to develop a better model system for retrovirus-induced disease.

为了开发逆转录病毒引起的疾病的基因治疗模型，将赋予宿主对小鼠白血病病毒（MuLV）感染具有强抵抗力的Fv-4抗性（Fv-4^r）基因引入到Friend白血病病毒（FLV）易感的C3H小鼠骨髓细胞中。1）Fv-4^r转染的骨髓的骨髓移植 细胞至C3H宿主使用SFFV载体系统引入Fv-4^r基因后，将C3H骨髓细胞移植至经致命性照射的C3H宿主中以产生骨髓嵌合体（Fv-4^r-G3H * C3H）。 Fv-4^r基因产物在这些嵌合体小鼠的造血细胞的细胞表面上表达。表达强度因小鼠而异，大部分表达量为携带Fv-4^r的C4W小鼠造血细胞表达量的10％至20％。2）高效基因诱导系统的开发通过选择载体系统和培养条件，我们可以产生Fv-4^r-C3H * C3H嵌合体，其Fv-4^r表达量超过C4W小鼠的30％ 3) 测试这些嵌合体对FLV感染的抵抗力的试验实验一些高表达Fv-4^r（> 30% of C4W）的Fv-4^r-C3H * C3H嵌合体表现出对1x103 PFU的FLV接种的抵抗力。然而，所有小鼠在感染lx 104 PFU的FLV后死亡。通过使用Fr-4^r-G3H * 高表达Fv-4^r的C3H嵌合体在最新实验中获得成功，我们将能够开发出更好的逆转录病毒诱导疾病模型系统。

项目成果

Kitagawa M et al.: "Overexpression of tumor necrosis factor(TNF)-α and interferon(IFN)-γ by bone marrow cells from patients with myelodysplastic syndromes" Leukemia. 11. 2049-2054 (1997)

Kitakawa M 等人：“骨髓增生异常综合征患者的骨髓细胞过度表达肿瘤坏死因子 (TNF)-α 和干扰素 (IFN)-γ”白血病。11. 2049-2054 (1997)

Kitagawa M et al.: "Cell-free transmission of Fv-4 resistance gene product controlling Friend leukemia virus-induced leukemigenesis in mice" Leukemia. 11 Suppl 3. 230-232 (1997)

Kitakawa M 等人：“Fv-4 抗性基因产物的无细胞传递控制 Friend 白血病病毒诱导的小鼠白血病发生”白血病。

Kitagawa M et al.: "Overexpression of tumor necrosis factor(TNF)-α and interferon(IFN)- γ by bone marrow cells from patients with myelodysplastic syndromes" Leukemia. 11. 2049-2054 (1997)

Kitakawa M 等人：“骨髓增生异常综合征患者的骨髓细胞过度表达肿瘤坏死因子 (TNF)-α 和干扰素 (IFN)-γ”白血病。11. 2049-2054 (1997)

Kamisaku H, Kitazawa M.et al.: "Limoithig diansmission analysis of T-cell progenitors in the bone of thymic thymic lymphome-susceptible BIO and-resistant C3H mice after fractionated whole-body X-irradiation" Int.J.Radiat.Biol.72. 191-199 (1997)

Kamisaku H、Kitazawa M.等人：“分次全身 X 射线照射后胸腺淋巴瘤敏感 BIO 和耐药 C3H 小鼠骨中 T 细胞祖细胞的 Limoithig 释放分析” Int.J.Radiat.Biol

Kitagawa M et al.: "Overexpression of tumor necrosis factor (TNF)-alpha and interferon (IFN) -gamma by bone marrow cells from patients with myelodysplastic syndromes" Leukemia. 11. 2049-2054 (1997)

Kitakawa M 等人：“骨髓增生异常综合征患者的骨髓细胞过度表达肿瘤坏死因子 (TNF)-α 和干扰素 (IFN)-γ”白血病。