GMP-compliant, large-scale expanded allogeneic natural killer cells have potent cytolytic activity against cancer cells in vitro and in vivo
Okjae Lim, Yuna Lee, Hyejin Chung, Jung Hyun Her, Sang Mi Kang, Mi-young Jung, Bokyung Min, Hyejin Shin, Tae Min Kim, Dae Seog Heo, Yu Kyeong Hwang, Eui-Cheol Shin, Okjae Lim, Yuna Lee, Hyejin Chung, Jung Hyun Her, Sang Mi Kang, Mi-young Jung, Bokyung Min, Hyejin Shin, Tae Min Kim, Dae Seog Heo, Yu Kyeong Hwang, Eui-Cheol Shin
Abstract
Ex vivo-expanded, allogeneic natural killer (NK) cells can be used for the treatment of various types of cancer. In allogeneic NK cell therapy, NK cells from healthy donors must be expanded in order to obtain a sufficient number of highly purified, activated NK cells. In the present study, we established a simplified and efficient method for the large-scale expansion and activation of NK cells from healthy donors under good manufacturing practice (GMP) conditions. After a single step of magnetic depletion of CD3(+) T cells, the depleted peripheral blood mononuclear cells (PBMCs) were stimulated and expanded with irradiated autologous PBMCs in the presence of OKT3 and IL-2 for 14 days, resulting in a highly pure population of CD3(-)CD16(+)CD56(+) NK cells which is desired for allogeneic purpose. Compared with freshly isolated NK cells, these expanded NK cells showed robust cytokine production and potent cytolytic activity against various cancer cell lines. Of note, expanded NK cells selectively killed cancer cells without demonstrating cytotoxicity against allogeneic non-tumor cells in coculture assays. The anti-tumor activity of expanded human NK cells was examined in SCID mice injected with human lymphoma cells. In this model, expanded NK cells efficiently controlled lymphoma progression. In conclusion, allogeneic NK cells were efficiently expanded in a GMP-compliant facility and demonstrated potent anti-tumor activity both in vitro and in vivo.
Conflict of interest statement
Competing Interests: Eui-Cheol Shin is a PLOS ONE Editorial Board member, and understands that this does not alter the authors' adherence to all the PLOS ONE policies on sharing data and materials. The authors have read the journal's policy and have the following conflicts: MJ and HS are employed in Green Cross LabCell Corp. Mogam Biotechnology Research Institute is a non-profit research foundation. Patent Number: KR2008-74069. Date Patent Issued: March 28, 2012. Title of Patent: Growth method for natural killer cells. Assignee: Green Cross LabCell Corp. and Seoul National University Hospital. Inventor: Mi-young Jung, Dae Seog Heo, and Yu Kyeong Hwang. Patent Number: JP2011-521023. Date Patent Filed: January 31, 2011. Title of Patent: Growth method for natural killer cells. Assignee: Green Cross LabCell Corp. and Seoul National University Hospital. Inventor: Mi-young Jung, Dae Seog Heo, and Yu Kyeong Hwang. Patent Number: CN200980130121.5. Date Patent Filed: January 30, 2011. Title of Patent: Growth method for natural killer cells. Assignee: Green Cross LabCell Corp. and Seoul National University Hospital. Inventor: Mi-young Jung, Dae Seog Heo, and Yu Kyeong Hwang. This does not alter the authors' adherence to all the PLOS ONE policies on sharing data and materials. The other authors declare that they have no conflict of interest.
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Source: PubMed