Sudarshawn N Damodharan1, Kirsti L Walker1, Matthew H Forsberg1, Kimberly A McDowell1, Myriam N Bouchlaka1, Diana A Drier2, Paul M Sondel3, Kenneth B DeSantes4, Christian M Capitini5. 1. Department of Pediatrics, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin, USA. 2. Waisman Biomanufacturing, University of Wisconsin, Madison, Wisconsin, USA. 3. Department of Pediatrics, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin, USA; Department of Human Oncology, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin, USA; Carbone Cancer Center, University of Wisconsin, Madison, Wisconsin, USA. 4. Department of Pediatrics, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin, USA; Carbone Cancer Center, University of Wisconsin, Madison, Wisconsin, USA. 5. Department of Pediatrics, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin, USA; Carbone Cancer Center, University of Wisconsin, Madison, Wisconsin, USA. Electronic address: ccapitini@pediatrics.wisc.edu.
Abstract
BACKGROUND AIMS: Several methods to expand and activate (EA) NK cells ex vivo have been developed for the treatment of relapsed or refractory cancers. Infusion of fresh NK cells is generally preferred to the infusion of cryopreserved/thawed (C/T) NK cells because of concern that cryopreservation diminishes NK cell activity. However, there has been little head-to-head comparison of the functionality of fresh versus C/T NK cell products. METHODS: We evaluated activity of fresh and C/T EA NK cells generated by interleukin (IL)-15, IL-2 and CD137L expansion. RESULTS: Analysis of C/T NK cell products demonstrated decreased recovery of viable CD56+ cells, but the proportion of NK cells in the C/T EA NK cell product did not decrease compared with the fresh EA NK cell product. Fresh and C/T EA NK cells demonstrated increased granzyme B compared with NK cells pre-expansion, but only fresh EA NK cells showed increased NKG2D. Compared with fresh EA NK cells, cytotoxic ability of C/T EA NK cells was reduced, but C/T EA NK cells remained potently cytotoxic against tumor cells via both antibody-independent and antibody-dependent mechanisms within 4 h post-thaw. Fresh EA NK cells generated high levels of gamma interferon (IFN-γ), which was abrogated by JAK1/JAK2 inhibition with ruxolitinib, but C/T EA NK cells showed lower IFN-γ unaffected by JAK1/JAK2 inhibition. DISCUSSION: Usage of C/T EA NK cells may be an option to provide serial "boost" NK cell infusions from a single apheresis to maximize NK cell persistence and potentially improve NK-induced responses to refractory cancer.
BACKGROUND AIMS: Several methods to expand and activate (EA) NK cells ex vivo have been developed for the treatment of relapsed or refractory cancers. Infusion of fresh NK cells is generally preferred to the infusion of cryopreserved/thawed (C/T) NK cells because of concern that cryopreservation diminishes NK cell activity. However, there has been little head-to-head comparison of the functionality of fresh versus C/T NK cell products. METHODS: We evaluated activity of fresh and C/T EA NK cells generated by interleukin (IL)-15, IL-2 and CD137L expansion. RESULTS: Analysis of C/T NK cell products demonstrated decreased recovery of viable CD56+ cells, but the proportion of NK cells in the C/T EA NK cell product did not decrease compared with the fresh EA NK cell product. Fresh and C/T EA NK cells demonstrated increased granzyme B compared with NK cells pre-expansion, but only fresh EA NK cells showed increased NKG2D. Compared with fresh EA NK cells, cytotoxic ability of C/T EA NK cells was reduced, but C/T EA NK cells remained potently cytotoxic against tumor cells via both antibody-independent and antibody-dependent mechanisms within 4 h post-thaw. Fresh EA NK cells generated high levels of gamma interferon (IFN-γ), which was abrogated by JAK1/JAK2 inhibition with ruxolitinib, but C/T EA NK cells showed lower IFN-γ unaffected by JAK1/JAK2 inhibition. DISCUSSION: Usage of C/T EA NK cells may be an option to provide serial "boost" NK cell infusions from a single apheresis to maximize NK cell persistence and potentially improve NK-induced responses to refractory cancer.
Authors: Melissa A Geller; Sarah Cooley; Patricia L Judson; Rahel Ghebre; Linda F Carson; Peter A Argenta; Amy L Jonson; Angela Panoskaltsis-Mortari; Julie Curtsinger; David McKenna; Kathryn Dusenbery; Robin Bliss; Levi S Downs; Jeffrey S Miller Journal: Cytotherapy Date: 2010-09-20 Impact factor: 5.414
Authors: Rizwan Romee; Maximillian Rosario; Melissa M Berrien-Elliott; Julia A Wagner; Brea A Jewell; Timothy Schappe; Jeffrey W Leong; Sara Abdel-Latif; Stephanie E Schneider; Sarah Willey; Carly C Neal; Liyang Yu; Stephen T Oh; Yi-Shan Lee; Arend Mulder; Frans Claas; Megan A Cooper; Todd A Fehniger Journal: Sci Transl Med Date: 2016-09-21 Impact factor: 17.956
Authors: Harry Dolstra; Mieke W H Roeven; Jan Spanholtz; Basav N Hangalapura; Marleen Tordoir; Frans Maas; Marij Leenders; Fenna Bohme; Nina Kok; Carel Trilsbeek; Jos Paardekooper; Anniek B van der Waart; Peter E Westerweel; Tjeerd J F Snijders; Jan Cornelissen; Gerard Bos; Hans F M Pruijt; Aniek O de Graaf; Bert A van der Reijden; Joop H Jansen; Arnold van der Meer; Gerwin Huls; Jeannette Cany; Frank Preijers; Nicole M A Blijlevens; Nicolaas M Schaap Journal: Clin Cancer Res Date: 2017-03-09 Impact factor: 12.531
Authors: Brian C Shaffer; Jean-Benoit Le Luduec; Christopher Forlenza; Ann A Jakubowski; Miguel-Angel Perales; James W Young; Katharine C Hsu Journal: Biol Blood Marrow Transplant Date: 2016-01-06 Impact factor: 5.742
Authors: Antonio Pérez-Martínez; Lucía Fernández; Jaime Valentín; Isabel Martínez-Romera; María Dolores Corral; Manuel Ramírez; Lorea Abad; Sandra Santamaría; Marta González-Vicent; Sara Sirvent; Julián Sevilla; José Luis Vicario; Inmaculada de Prada; Miguel Ángel Diaz Journal: Cytotherapy Date: 2015-09-01 Impact factor: 5.414
Authors: Caroline J Voskens; Ryuko Watanabe; Sandra Rollins; Dario Campana; Kenichiro Hasumi; Dean L Mann Journal: J Exp Clin Cancer Res Date: 2010-10-11
Authors: Carlos E Sanchez; Ehsan P Dowlati; Ashley E Geiger; Kajal Chaudhry; Matthew A Tovar; Catherine M Bollard; Conrad Russell Y Cruz Journal: Transplant Cell Ther Date: 2020-09-29
Authors: Jeremiah L Oyer; Tayler J Croom-Perez; Thomas A Dieffenthaller; Liza D Robles-Carillo; Sarah B Gitto; Deborah A Altomare; Alicja J Copik Journal: Front Immunol Date: 2022-04-07 Impact factor: 8.786