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为研究基因治疗在造血功能损伤后恢复中的应用,本课题以携带小鼠IL-3基因的复制缺陷型重组腺病毒载体转染骨髓基质细胞,对大剂量化疗后的小鼠进行脾内移植观察造血功能的恢复情况.结果表明,缺陷型腺病毒载体能有效地转染小鼠原代骨髓基质细胞,转染效率在80%以上(MOI=10);基因修饰的骨髓基质细胞体外分泌IL-3的水平可达110U/ml/10~6细胞/24小时;在大剂量环磷酰胺治疗后脾内移植IL-3基因修饰的基质细胞能有效地升高实验小鼠外周血白细胞总数;病理检测发现IL-3基因修饰的基质细胞治疗组小鼠脾脏和骨髓中细胞增生较其它组明显活跃;经IL-3基因修饰的基质细胞治疗组小鼠脾淋巴细胞对ConA反应明显增强.结果提示IL-3基因修饰的骨髓基质细胞体内移植对大剂量化疗后机体造血与免疫功能的恢复都有较好的促进作用.
In order to study the application of gene therapy in recovery after hematopoietic damage, this study transfected bone marrow stromal cells with a replication-defective recombinant adenovirus vector carrying the mouse IL-3 gene and performed intrasplenic transplantation of mice after high-dose chemotherapy. The recovery of hematopoietic function was observed. The results showed that the defective adenovirus vector can effectively transfect mouse primary bone marrow stromal cells with a transfection efficiency of more than 80% (MOI=10); genetically modified bone marrow stromal cells excrete IL in vitro The level of -3 can reach 110U/ml/10~6 cells/24 hours. Transplantation of IL-3 gene-modified stromal cells into the spleen after high-dose cyclophosphamide treatment can effectively increase the total number of white blood cells in the peripheral blood of the experimental mice. Pathological examination showed that cell proliferation in the spleen and bone marrow of mice treated with IL-3 gene-modified stromal cells was significantly more active than that of other groups; the response of ConA to spleen lymphocytes was significantly enhanced in mice treated with IL-3 gene-modified stromal cells. It suggests that the in vivo transplantation of IL-3 gene-modified bone marrow stromal cells has a good promoting effect on the recovery of hematopoiesis and immune function after high-dose chemotherapy.