Fehse B, Kustikova O S, Li Z, Wahlers A, Bohn W, Beyer W R, Chalmers D, Tiberghien P, Kühlcke K, Zander A R, Baum C
Bone Marrow Transplantation, University Hospital Hamburg-Eppendorf, Germany.
Gene Ther. 2002 Dec;9(23):1633-8. doi: 10.1038/sj.gt.3301828.
Retroviral suicide gene vectors have successfully been used in clinical studies to improve the safety of adoptive immunotherapy with allogeneic T lymphocytes in the treatment of malignant and viral diseases. At the same time these studies have revealed several problems that are yet to be resolved including impaired T cell function due to long ex vivo culture. Here we present new retroviral vectors co-expressing truncated CD34, a gene transfer marker which ensures rapid enrichment of transduced cells using commercially available GMP-approved devices, and a splice-corrected variant of Herpes simplex virus thymidine kinase (scHSVtk) which confers high sensitivity to the prodrug ganciclovir. We show that a retroviral hybrid vector, MP71, based on the myeloproliferative sarcoma virus (MPSV) and the murine embryonic stem cell virus (MESV), encoding a tCD34/scHSVtk fusion protein mediates high expression of the 'sort-suicide' selection marker, thereby allowing for highly efficient purification and selective elimination of transduced cells.
逆转录病毒自杀基因载体已成功应用于临床研究，以提高在恶性疾病和病毒性疾病治疗中采用同种异体T淋巴细胞进行过继性免疫治疗的安全性。与此同时，这些研究也揭示了一些尚待解决的问题，包括由于长时间体外培养导致的T细胞功能受损。在此，我们展示了新的逆转录病毒载体，其共表达截短的CD34（一种基因转移标记，可使用市售的符合GMP标准的设备确保对转导细胞进行快速富集）和单纯疱疹病毒胸苷激酶的剪接校正变体（scHSVtk），该变体对前体药物更昔洛韦具有高敏感性。我们表明，基于骨髓增殖性肉瘤病毒（MPSV）和小鼠胚胎干细胞病毒（MESV）的逆转录病毒杂交载体MP71，编码tCD34/scHSVtk融合蛋白，介导“分选-自杀”选择标记的高表达，从而允许对转导细胞进行高效纯化和选择性清除。
