Graduate School of Anhui Medical University, 81 Meishan Road, Shu Shan Qu, Hefei, Anhui, People's Republic of China.
Department of Experimental Hematology, Beijing Institute of Radiation Medicine, 27 Taiping Road, Beijing, China.
BMC Biotechnol. 2019 Apr 24;19(1):23. doi: 10.1186/s12896-019-0514-x.
The gene transduction efficiency of adenovirus to hematopoietic cells, especially T lymphocytes, is needed to be improved. The purpose of this study is to improve the transduction efficiency of T lymphocytes by using fiber-modified human adenovirus 5 (HAdV-5) vectors.
Four fiber-modified human adenovirus 5 (HAdV-5) vectors were investigated to transduce hematopoietic cells. F35-EG or F11p-EG were HAdV-35 or HAdV-11p fiber pseudotyped HAdV-5, and HR-EG or CR-EG vectors were generated by incorporating RGD motif to the HI loop or to the C-terminus of F11p-EG fiber. All vectors could transduce more than 90% of K562 or Jurkat cells at an multiplicity of infection (MOI) of 500 viral particle per cell (vp/cell). All vectors except HR-EG could transduce nearly 90% cord blood CD34+ cells or 80% primary human T cells at the MOI of 1000, and F11p-EG showed slight superiority to F35-EG and CR-EG. Adenoviral vectors transduced CD4+ T cells a little more efficiently than they did to CD8+ T cells. These vectors showed no cytotoxicity at an MOI as high as 1000 vp/cell because the infected and uninfected T cells retained the same CD4/CD8 ratio and cell growth rate.
HAdV-11p fiber pseudotyped HAdV-5 could effectively transduce human T cells when human EF1a promoter was used to control the expression of transgene, suggesting its possible application in T cell immunocellular therapy.
需要提高腺病毒对造血细胞(尤其是 T 淋巴细胞)的基因转导效率。本研究旨在通过使用纤维修饰的人腺病毒 5 型(HAdV-5)载体来提高 T 淋巴细胞的转导效率。
研究了四种纤维修饰的人腺病毒 5 型(HAdV-5)载体来转导造血细胞。F35-EG 或 F11p-EG 是 HAdV-35 或 HAdV-11p 纤维假型 HAdV-5,而 HR-EG 或 CR-EG 载体是通过将 RGD 基序插入 HI 环或 F11p-EG 纤维的 C 末端而产生的。所有载体在感染复数(MOI)为 500 个病毒颗粒/细胞(vp/cell)时,均能转导超过 90%的 K562 或 Jurkat 细胞。除 HR-EG 外,所有载体在 MOI 为 1000 时均能转导近 90%的脐血 CD34+细胞或 80%的原代人 T 细胞,且 F11p-EG 比 F35-EG 和 CR-EG 略有优势。腺病毒载体转导 CD4+T 细胞的效率略高于 CD8+T 细胞。当使用人 EF1a 启动子来控制转基因的表达时,这些载体在高达 1000 vp/cell 的 MOI 下没有细胞毒性,因为感染和未感染的 T 细胞保持相同的 CD4/CD8 比值和细胞生长速度。
当使用人 EF1a 启动子控制转基因的表达时,HAdV-11p 纤维假型 HAdV-5 可有效地转导人 T 细胞,提示其在 T 细胞免疫细胞治疗中的可能应用。