Ding L, Lu S, Batchu R B, Saylors R L, Munshi N C
Central Arkansas Veterans Healthcare System, Myeloma and Transplantation Research Center, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA.
Gene Ther. 1999 Sep;6(9):1611-6. doi: 10.1038/sj.gt.3300973.
Adoptive transfer of genetically modified somatic cells is playing an increasingly important role in the management of a wide spectrum of human diseases. Hematopoietic stem cells and lymphocytes have been used to transfer a variety of genes, however, they have limitations. In this study, the feasibility of retroviral gene transduction of bone marrow stromal cells, and the engraftment characteristics of these cells following infusion, was investigated in a murine transplantation model. Stromal cells derived from Balb/c mouse bone marrow were transduced with a replication-defective retrovirus containing the LacZ gene. Following three rounds of transduction, between 5 and 40% of the cells were positive for the LacZ gene. A total of 2 x 106 cells were infused into the same mouse strain. After the infusion, the LacZ gene was detected by PCR in the bone marrow, spleen, liver, kidney and lung; however, only the spleen and bone marrow samples were strongly positive. Quantitative PCR demonstrated that between 3 and 5% of spleen and bone marrow cells, and 1% of liver cells contained the LacZ gene at 3 weeks after infusion; <0.2% transduced cells were found in other organs. No difference was noted in engraftment between mice with or without irradiation before transplantation, suggesting that engraftment occurred without myeloablation. The infused transduced cells persisted for up to 24 weeks. Self-renewal of transplanted stromal cells was demonstrated in secondary transplant studies. Ease of culture and gene transduction and tissue specificity to hematopoietic organs (bone marrow, spleen, liver) is demonstrated, indicating that stromal cells may be an ideal vehicle for gene transfer.
基因修饰体细胞的过继转移在多种人类疾病的治疗中发挥着越来越重要的作用。造血干细胞和淋巴细胞已被用于转移多种基因，然而，它们存在局限性。在本研究中，在小鼠移植模型中研究了逆转录病毒基因转导骨髓基质细胞的可行性，以及输注后这些细胞的植入特性。用含有LacZ基因的复制缺陷型逆转录病毒转导来自Balb/c小鼠骨髓的基质细胞。经过三轮转导后，5%至40%的细胞LacZ基因呈阳性。将总共2×10⁶个细胞输注到同一小鼠品系中。输注后，通过PCR在骨髓、脾脏、肝脏、肾脏和肺中检测到LacZ基因；然而，只有脾脏和骨髓样本呈强阳性。定量PCR表明，输注后3周，脾脏和骨髓细胞中有3%至5%，肝细胞中有1%含有LacZ基因；在其他器官中发现的转导细胞<0.2%。移植前接受或未接受照射的小鼠在植入方面没有差异，这表明植入发生在未进行清髓的情况下。输注的转导细胞持续存在长达24周。二次移植研究证明了移植的基质细胞具有自我更新能力。证明了培养和基因转导的便利性以及对造血器官（骨髓、脾脏、肝脏）的组织特异性，表明基质细胞可能是基因转移的理想载体。
