Department of Pediatrics, University of Tsukuba Hospital, 2-1-1 Amakubo, Tsukuba, Ibaraki, 305-8576, Japan.
Department of Child Health, Faculty of Medicine, University of Tsukuba, Ibaraki, Japan.
Int J Hematol. 2021 Apr;113(4):493-499. doi: 10.1007/s12185-020-03059-6. Epub 2021 Jan 1.
Sendai virus (SeV) vectors are being recognized as a superior tool for gene transfer. Here, we report the transfection efficacy of a novel, high-performance, replication-defective, and persistent Sendai virus (SeVdp) vector in cultured cells and in mice using a near-infrared fluorescent protein (iRFP)-mediated in vivo imaging system. The novel SeVdp vector established persistent infection, and strong expression of inserted genes was sustained indefinitely in vitro. Analysis of iRFP-expressing cells transplanted subcutaneously into NOG, nude, and ICR mice suggests that innate immunity was involved in the exclusion of the transplanted cells. We also evaluated the feasibility of this novel SeVdp vector for hemophilia A gene therapy. This system enabled insertion of full-length FVIII genes, and transduced cells secreted FVIII into the culture medium. Transient FVIII activity was detected in the plasma of mice after intraperitoneal transplantation of these FVIII-secreting cells. Further improvement in methods to evade immunity, such as simultaneous expression of immunomodulatory genes, would make this novel vector a very useful tool in regenerative medicine.
仙台病毒（SeV）载体正被认为是基因转移的一种优越工具。在这里，我们报告了一种新型的、高性能的、复制缺陷的和持续存在的仙台病毒（SeVdp）载体在培养细胞中和使用近红外荧光蛋白（iRFP）介导的体内成像系统的小鼠中的转染效率。新型 SeVdp 载体建立了持续感染，并且插入基因的强表达在体外无限期地持续。对皮下移植到 NOG、裸鼠和 ICR 小鼠中的 iRFP 表达细胞的分析表明，固有免疫参与了移植细胞的排斥。我们还评估了这种新型 SeVdp 载体用于血友病 A 基因治疗的可行性。该系统能够插入全长 FVIII 基因，并且转导的细胞将 FVIII 分泌到培养基中。在这些分泌 FVIII 的细胞经腹腔移植后，在小鼠的血浆中检测到短暂的 FVIII 活性。进一步改进逃避免疫的方法，例如同时表达免疫调节基因，将使这种新型载体成为再生医学中非常有用的工具。
