Genetic modification of human T cells with CD20: A strategy to purify and lyse transduced cells with anti-CD20 antibodies
Genetic modification of human T cells with CD20: A strategy to purify and lyse transduced cells with anti-CD20 antibodies
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DOI：
10.1089/10430340050015798
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发表时间：
2000-03-01
影响因子：
4.2
通讯作者：
Rambaldi, A
中科院分区：
文献类型：
--
作者：
Introna, M;Barbui, AM;Rambaldi, A
已构建了一种逆转录病毒载体，其中包含受莫洛尼鼠白血病病毒 (Mo-MuLV) LTR 控制的人 CD20 cDNA。新鲜分离的单核细胞在 PHA 和 hrIL-2 存在下连续感染三天，平均 15.9% 的细胞（范围为 6.5 至 31.7%）获得 CD3(+)CD20(+) 表型。转导的 T 淋巴细胞像模拟感染的细胞一样在体外生长和扩增长达 3 周，并且正如在 T 淋巴母细胞 CEM 细胞系中观察到的那样，CD20 表达可维持数月，细胞生长曲线没有变化。使用不同的抗 CD20 抗体，可以在柱上对表达 CD20 的 CEM 和新鲜 T 淋巴细胞进行阳性免疫选择。在补体存在下，接触单克隆嵌合抗 CD20 IgG(1)(kappa) Rituximab 抗体 (Roche) 可在体外有效、快速地杀死转导的 CD3(+)CD20(+) 人 T 细胞。这种方法代表了一种新的、替代性的基因操作方法，用“自杀”基因来产生药物反应性 T 细胞群，这是未来管理骨髓移植患者移植物抗宿主病的关键一步。
A retroviral vector has been constructed that contains the human CD20 cDNA under the control of the Moloney murine leukemia virus (Mo-MuLV) LTR. Freshly isolated mononuclear cells are infected for three consecutive days in the presence of PHA and hrIL-2 and a mean 15.9% of the cells (range, 6.5 to 31.7%) acquire a CD3(+)CD20(+) phenotype. Transduced T lymphocytes grow and expand in vitro for up to 3 weeks like mock-infected cells and, as observed for the T lymphoblastoid CEM cell line, CD20 expression is maintained for several months with no change in the growth curve of the cells. CD20-expressing CEM and fresh T lymphocytes can be positively immunoselected on columns using different anti-CD20 antibodies. Exposure to monoclonal chimeric anti-CD20 IgG(1)(kappa) Rituximab antibody (Roche), in the presence of complement, results in effective and rapid killing of the transduced CD3(+)CD20(+) human T cells in vitro. This approach represents a new and alternative method to gene manipulation with "suicide" genes for the production of drug-responsive T cell populations, a crucial step for the future management of graft-versus-host disease in bone marrow transplant patients.