Expression of chimeric antigen receptors in natural killer cells with a regulatory-compliant non-viral method
| Autor: | Madhusudan V. Peshwa, Cornell Allen, Stephanie Feller, Sergey Dzekunov, Hiroyuki Fujisaki, Lawrence A. Wolfraim, Linhong Li, Joseph C. Fratantoni, Linda N. Liu, Dario Campana, Rama Shivakumar, Nicholas Chopas |
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| Rok vydání: | 2009 |
| Předmět: |
Cancer Research
Cell Survival Antigens CD19 Green Fluorescent Proteins Transfection Article CD19 Cell Line Immunophenotyping Interleukin 21 Cell Line Tumor Humans Antigen-presenting cell Molecular Biology Cells Cultured Lymphokine-activated killer cell biology Janus kinase 3 fungi Natural killer T cell Molecular biology Cell biology Killer Cells Natural Receptors Antigen Electroporation Interleukin 12 biology.protein Molecular Medicine |
| Zdroj: | Cancer Gene Therapy. 17:147-154 |
| ISSN: | 1476-5500 0929-1903 |
| DOI: | 10.1038/cgt.2009.61 |
| Popis: | Natural killer (NK) 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 h 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 > or =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 h 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. |
| Databáze: | OpenAIRE |
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