Newrzela Sebastian, Gunda Brandenburg, von Laer Dorothee
Institute for Biomedical Research, Georg-Speyer-Haus, Frankfurt am Main, Germany.
Methods Mol Biol. 2009;506:71-82. doi: 10.1007/978-1-59745-409-4_6.
Gene transfer into mature T cells with gammaretroviral vectors requires prestimulation, as only mitotic cells are susceptible to integration of the gammaretroviral proviral genome. Costimulation via the CD3/ TCR complex and a second costimulatory molecule, such as CD28 was found to better preserve functionality of the T lymphocytes during ex vivo expansion than stimulation with anti-CD3 alone. The protocols described here for prestimulation and transduction of human and murine T cells with gammaretroviral vectors were optimized for high-level gene transfer and maximum yield of functional T lymphocytes.
使用γ逆转录病毒载体将基因导入成熟T细胞需要预先刺激，因为只有有丝分裂细胞才易受γ逆转录病毒前病毒基因组整合的影响。与单独用抗CD3刺激相比，经由CD3/TCR复合体和第二种共刺激分子（如CD28）进行共刺激，在体外扩增过程中能更好地保留T淋巴细胞的功能。此处所述的用γ逆转录病毒载体对人和小鼠T细胞进行预先刺激和转导的方案，针对高水平基因转移和功能性T淋巴细胞的最大产量进行了优化。
