Attempt for gene therapy of retrovirus-induced disease

为了建立逆转录病毒诱导疾病的基因治疗模型，将具有抗小鼠白血病病毒（MuLV）感染能力的Fv-4耐药基因（Fv-4^r）引入Friend白血病病毒（FLV）易感的C3H小鼠骨髓细胞。1)将转染Fv-4^r的骨髓细胞移植到C3H宿主体内，利用SFFV载体系统导入Fv-4^r基因后，将C3H骨髓细胞移植到经超致死照射的C3H宿主体内，形成骨髓嵌合体（Fv-4^r- g3h * C3H）。Fv-4^r基因产物在这些嵌合体小鼠造血细胞表面表达。不同小鼠的表达强度不同，大多数小鼠的造血细胞表达Fv-4^r基因的C4W小鼠的表达量在10%到20%之间。2)建立高效的基因诱导体系通过选择载体体系和培养条件，我们可以产生Fv-4^r-C3H * C3H嵌合体，Fv-4^r的表达量超过C4W小鼠的30%。少量高表达Fv-4^r（约占C4W的30%）的Fv-4^r * C3H嵌合体对接种1 × 103 PFU的FLV具有抗性。然而，所有小鼠在感染lx104 PFU的FLV后死亡。利用最新实验成功的高表达Fv-4^r的Fr-4^r- g3h * C3H嵌合体，我们可以建立一个更好的逆转录病毒诱导疾病的模型系统。

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.