Expression of chimeric antigen receptors in natural killer cells with a regulatory-compliant non-viral method.
Expression of chimeric antigen receptors in natural killer cells with a regulatory-compliant non-viral method.
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DOI：
10.1038/cgt.2009.61
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发表时间：
2010-03
影响因子：
6.4
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中科院分区：
文献类型：
--
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自然杀伤细胞为癌症治疗带来希望。NK细胞毒性可以通过嵌合抗原受体的表达来增强，所述嵌合抗原受体通过接合在靶细胞上表达的细胞表面分子而将特异性重新导向靶细胞。我们开发了一种符合法规的、可扩展的非病毒方法，以基于编码嵌合受体的mRNA的转染将NK细胞工程化为靶特异性的。将eGFP mRNA转染到离体扩增的NK细胞（N=5）或来自外周血的纯化的未刺激的NK细胞（N=4）中导致良好的细胞活力，转染后24小时，eGFP表达分别为85% ± 6%和86% ± 4%。编码针对CD 19的受体（抗-CD 19-BB-z）的mRNA也被有效地转染到NK细胞中。表达抗CD 19-BB-z的离体扩增和纯化的未刺激NK细胞对CD 19+靶细胞的细胞毒性增强，导致急性淋巴细胞白血病和B系慢性淋巴细胞白血病细胞裂解≥80%，效应靶比低于10：1。早在转染后3小时就观察到抗CD 19-BB-z mRNA转染的NK细胞的靶特异性细胞毒性，并持续长达3天。本文所述的方法应有助于基于NK的肿瘤抗原靶向免疫治疗的临床开发。
Natural killer cells hold promise for cancer therapy. NK cytotoxicity can be enhanced by expression of chimeric antigen receptors that re-direct specificity toward target cells by engaging cell surface molecules expressed on target cells. We developed a regulatory-compliant, scalable non-viral approach to engineer NK cells to be target-specific based on transfection of mRNA encoding chimeric receptors. Transfection of eGFP mRNA into ex vivo expanded NK cells (N=5) or purified unstimulated NK cells from peripheral blood (N=4) resulted in good cell viability with eGFP expression in 85% ± 6% and 86% ± 4%, 24 hours after transfection, respectively. An mRNA encoding a receptor directed against CD19 (anti-CD19-BB-z) was also transfected into NK cells efficiently. Ex vivo expanded and purified unstimulated NK cells expressing anti-CD19-BB-z exhibited enhanced cytotoxicity against CD19+ target cells resulting in ≥80% lysis of acute lymphoblastic leukemia and B-lineage chronic lymphocytic leukemia cells at effector target ratios lower than 10:1. The target-specific cytotoxicity for anti-CD19-BB-z mRNA-transfected NK cells was observed as early as 3 hours after transfection and persisted for up to 3 days. The method described here should facilitate the clinical development of NK-based antigen-targeted immunotherapy for cancer.
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影响因子：
20.3
作者：
Holtkamp, Silke;Kreiter, Sebastian;Sahin, Ugur
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作者：
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