An efficient feeder-free and chemically-defined expansion strategy for highly purified natural killer cells derived from human cord blood
Introduction: Natural killer cells (NKCs) are immune cells that can attack cancer cells through the direct recognition of ligands without prior sensitization. Cord blood-derived NKCs (CBNKCs) represent a promising tool for allogenic NKC-based cancer immunotherapy. Efficient NKC expansion and decreas...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-12-01
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| Series: | Regenerative Therapy |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2352320423000445 |
| _version_ | 1797448066818637824 |
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| author | Tsutomu Nakazawa Ryosuke Maeoka Takayuki Morimoto Ryosuke Matsuda Mitsutoshi Nakamura Fumihiko Nishimura Shuichi Yamada Ichiro Nakagawa Young-Soo Park Toshihiro Ito Hiroyuki Nakase Takahiro Tsujimura |
| author_facet | Tsutomu Nakazawa Ryosuke Maeoka Takayuki Morimoto Ryosuke Matsuda Mitsutoshi Nakamura Fumihiko Nishimura Shuichi Yamada Ichiro Nakagawa Young-Soo Park Toshihiro Ito Hiroyuki Nakase Takahiro Tsujimura |
| author_sort | Tsutomu Nakazawa |
| collection | DOAJ |
| description | Introduction: Natural killer cells (NKCs) are immune cells that can attack cancer cells through the direct recognition of ligands without prior sensitization. Cord blood-derived NKCs (CBNKCs) represent a promising tool for allogenic NKC-based cancer immunotherapy. Efficient NKC expansion and decreased T cell inclusion are crucial for the success of allogeneic NKC-based immunotherapy without inducing graft-versus-host reactions. We previously established an efficient ex vivo expansion system consisting of highly purified-NKCs derived from human peripheral blood. Herein, we evaluated the performance of the NKC expansion system using CB and characterized the expanded populations. Methods: Frozen CB mononuclear cells (CBMCs), with T cells removed, were cultured with recombinant human interleukin (rhIL)-18 and rhIL-2 under conditions where anti-NKp46 and anti-CD16 antibodies were immobilized. Following 7, 14, and 21 days of expansion, the purity, fold-expansion rates of NKCs, and the expression levels of NK activating and inhibitory receptors were assessed. The ability of these NKCs to inhibit the growth of T98G, a glioblastoma (GBM) cell line sensitive to NK activity, was also examined. Results: All expanded T cell-depleted CBMCs were included in over 80%, 98%, and 99% of CD3−CD56+ NKCs at 7, 14, and 21 days of expansion, respectively. The NK activating receptors LFA-1, NKG2D, DNAM-1, NKp30, NKp44, NKp46, FcγRIII and NK inhibitory receptors TIM-3, TIGIT, TACTILE, NKG2A were expressed on the expanded-CBNKCs. Two out of three of the expanded-CBNKCs weakly expressed PD-1, yet gradually expressed PD-1 according to expansion period. One of the three expanded CBNKCs almost lacked PD-1 expression during the expansion period. LAG-3 expression was variable among donors, and no consistent changes were identified during the expansion period. All of the expanded CBNKCs elicited distinct cytotoxicity-mediated growth inhibition on T98G cells. The level of cytotoxicity was gradually decreased based on the prolonged expansion period. Conclusions: Our established feeder-free expansion system yielded large scale highly purified and cytotoxic NKCs derived from human CB. The system provides a stable supply of clinical grade off-the-shelf NKCs and may be feasible for allogeneic NKC-based immunotherapy for cancers, including GBM. |
| first_indexed | 2024-03-09T14:05:00Z |
| format | Article |
| id | doaj.art-99ed7a75bb9e402cb9fb37eedbaf9b59 |
| institution | Directory Open Access Journal |
| issn | 2352-3204 |
| language | English |
| last_indexed | 2024-03-09T14:05:00Z |
| publishDate | 2023-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Regenerative Therapy |
| spelling | doaj.art-99ed7a75bb9e402cb9fb37eedbaf9b592023-11-30T05:07:42ZengElsevierRegenerative Therapy2352-32042023-12-01243242An efficient feeder-free and chemically-defined expansion strategy for highly purified natural killer cells derived from human cord bloodTsutomu Nakazawa0Ryosuke Maeoka1Takayuki Morimoto2Ryosuke Matsuda3Mitsutoshi Nakamura4Fumihiko Nishimura5Shuichi Yamada6Ichiro Nakagawa7Young-Soo Park8Toshihiro Ito9Hiroyuki Nakase10Takahiro Tsujimura11Grandsoul Research Institute for Immunology, Inc., Uda, Nara, 633-2221, Japan; Clinic Grandsoul Nara, Matsui 8-1, Uda, Nara, 633-2221, Japan; Department of Neurosurgery, Nara Medical University, Kashihara, Nara, 634-8522, Japan; Corresponding author. Grandsoul Research Institute for Immunology, Inc., Matsui 8-1, Utano, Uda, Nara, 633-2221, Japan. Tel./fax: +81 745 84 9335.Department of Neurosurgery, Nara Medical University, Kashihara, Nara, 634-8522, JapanDepartment of Neurosurgery, Nara Medical University, Kashihara, Nara, 634-8522, JapanDepartment of Neurosurgery, Nara Medical University, Kashihara, Nara, 634-8522, JapanClinic Grandsoul Nara, Matsui 8-1, Uda, Nara, 633-2221, Japan; Department of Neurosurgery, Nara Medical University, Kashihara, Nara, 634-8522, JapanDepartment of Neurosurgery, Nara Medical University, Kashihara, Nara, 634-8522, JapanDepartment of Neurosurgery, Nara Medical University, Kashihara, Nara, 634-8522, JapanDepartment of Neurosurgery, Nara Medical University, Kashihara, Nara, 634-8522, JapanDepartment of Neurosurgery, Nara Medical University, Kashihara, Nara, 634-8522, JapanDepartment of Immunology, Nara Medical University, Kashihara, Nara, 634-8522, JapanDepartment of Neurosurgery, Nara Medical University, Kashihara, Nara, 634-8522, JapanGrandsoul Research Institute for Immunology, Inc., Uda, Nara, 633-2221, Japan; Clinic Grandsoul Nara, Matsui 8-1, Uda, Nara, 633-2221, JapanIntroduction: Natural killer cells (NKCs) are immune cells that can attack cancer cells through the direct recognition of ligands without prior sensitization. Cord blood-derived NKCs (CBNKCs) represent a promising tool for allogenic NKC-based cancer immunotherapy. Efficient NKC expansion and decreased T cell inclusion are crucial for the success of allogeneic NKC-based immunotherapy without inducing graft-versus-host reactions. We previously established an efficient ex vivo expansion system consisting of highly purified-NKCs derived from human peripheral blood. Herein, we evaluated the performance of the NKC expansion system using CB and characterized the expanded populations. Methods: Frozen CB mononuclear cells (CBMCs), with T cells removed, were cultured with recombinant human interleukin (rhIL)-18 and rhIL-2 under conditions where anti-NKp46 and anti-CD16 antibodies were immobilized. Following 7, 14, and 21 days of expansion, the purity, fold-expansion rates of NKCs, and the expression levels of NK activating and inhibitory receptors were assessed. The ability of these NKCs to inhibit the growth of T98G, a glioblastoma (GBM) cell line sensitive to NK activity, was also examined. Results: All expanded T cell-depleted CBMCs were included in over 80%, 98%, and 99% of CD3−CD56+ NKCs at 7, 14, and 21 days of expansion, respectively. The NK activating receptors LFA-1, NKG2D, DNAM-1, NKp30, NKp44, NKp46, FcγRIII and NK inhibitory receptors TIM-3, TIGIT, TACTILE, NKG2A were expressed on the expanded-CBNKCs. Two out of three of the expanded-CBNKCs weakly expressed PD-1, yet gradually expressed PD-1 according to expansion period. One of the three expanded CBNKCs almost lacked PD-1 expression during the expansion period. LAG-3 expression was variable among donors, and no consistent changes were identified during the expansion period. All of the expanded CBNKCs elicited distinct cytotoxicity-mediated growth inhibition on T98G cells. The level of cytotoxicity was gradually decreased based on the prolonged expansion period. Conclusions: Our established feeder-free expansion system yielded large scale highly purified and cytotoxic NKCs derived from human CB. The system provides a stable supply of clinical grade off-the-shelf NKCs and may be feasible for allogeneic NKC-based immunotherapy for cancers, including GBM.http://www.sciencedirect.com/science/article/pii/S2352320423000445Cord bloodAllogeneic NKCCell-based immunotherapyCancer immunotherapyGlioblastoma |
| spellingShingle | Tsutomu Nakazawa Ryosuke Maeoka Takayuki Morimoto Ryosuke Matsuda Mitsutoshi Nakamura Fumihiko Nishimura Shuichi Yamada Ichiro Nakagawa Young-Soo Park Toshihiro Ito Hiroyuki Nakase Takahiro Tsujimura An efficient feeder-free and chemically-defined expansion strategy for highly purified natural killer cells derived from human cord blood Regenerative Therapy Cord blood Allogeneic NKC Cell-based immunotherapy Cancer immunotherapy Glioblastoma |
| title | An efficient feeder-free and chemically-defined expansion strategy for highly purified natural killer cells derived from human cord blood |
| title_full | An efficient feeder-free and chemically-defined expansion strategy for highly purified natural killer cells derived from human cord blood |
| title_fullStr | An efficient feeder-free and chemically-defined expansion strategy for highly purified natural killer cells derived from human cord blood |
| title_full_unstemmed | An efficient feeder-free and chemically-defined expansion strategy for highly purified natural killer cells derived from human cord blood |
| title_short | An efficient feeder-free and chemically-defined expansion strategy for highly purified natural killer cells derived from human cord blood |
| title_sort | efficient feeder free and chemically defined expansion strategy for highly purified natural killer cells derived from human cord blood |
| topic | Cord blood Allogeneic NKC Cell-based immunotherapy Cancer immunotherapy Glioblastoma |
| url | http://www.sciencedirect.com/science/article/pii/S2352320423000445 |
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