Nayoun Kim1,2,3, Young-Sun Nam1,2, Keon-Il Im1,2, Jung-Yeon Lim1,2, Young-Woo Jeon1,2,4, Yunejin Song1,2, Jong Wook Lee4, Seok-Goo Cho1,2,3,4. 1. Institute for Translational Research and Molecular Imaging, The Catholic University of Korea College of Medicine, Seoul, South Korea. 2. Laboratory of Immune Regulation, Convergent Research Consortium for Immunologic Disease, Seoul, South Korea. 3. Catholic Institute of Cell TherapySeoul St. Mary's Hospital, The Catholic University of Korea College of Medicine, Seoul, South Korea. 4. Catholic Blood and Marrow Transplantation Center, Seoul St. Mary's Hospital, The Catholic University of Korea College of Medicine, Seoul, South Korea.
Abstract
BACKGROUND: Cytomegalovirus(CMV)-related diseases are a serious cause of morbidity and mortality following hematopoietic stem cell transplantation (HSCT). CMV-specific cytotoxic T lymphocytes (CMV-CTLs) have been reported as an alternative to antiviral drugs that provide long-term CMV-specific immunity without major side effects. However, their application has been limited by the prolonged manufacturing process required. METHODS: In this study, we applied the IFN-γ cytokine capture system (CCS) using the fully automated CliniMACS Prodigy device for rapid production of CMV-CTLs, which may be applicable in clinically urgent CMV-related diseases. Five validation runs were performed using apheresis samples from randomly selected CMV-seropositive healthy blood donors. Successive processes, including antigen stimulation, anti-IFN-γ labeling, magnetic enrichment and elution, were then performed automatically using the CliniMACS Prodigy, which took approximately 13 h. RESULTS: The original apheresis samples consisted mainly of CD45RA+ CD62L+ naïve T cells as well as 0.3% IFN-γ-secreting CD3+ T cells that showed a response to the CMV pp65 antigen (CD3+ IFN-γ+ cells). Following IFN-γ enrichment, the target fraction contained 51.3% CD3+ IFN-γ+ cells with a reduction in naïve T cells and selection of CD45RA- CD62L- and CD45RA+ CD62L- memory T cells. Furthermore, extended culture of these isolated cells revealed functional activity, including efficient proliferation, sustained antigen-specific IFN-γ secretion, and cytotoxicity against pp65-pulsed target cells. CONCLUSION: The findings reported here suggest that the IFN-γ CCS by the CliniMACS Prodigy is a simple and robust approach to produce CMV-CTLs, which may be applicable for the treatment of clinically urgent CMV-related diseases.
BACKGROUND: Cytomegalovirus(CMV)-related diseases are a serious cause of morbidity and mortality following hematopoietic stem cell transplantation (HSCT). CMV-specific cytotoxic T lymphocytes (CMV-CTLs) have been reported as an alternative to antiviral drugs that provide long-term CMV-specific immunity without major side effects. However, their application has been limited by the prolonged manufacturing process required. METHODS: In this study, we applied the IFN-γ cytokine capture system (CCS) using the fully automated CliniMACS Prodigy device for rapid production of CMV-CTLs, which may be applicable in clinically urgent CMV-related diseases. Five validation runs were performed using apheresis samples from randomly selected CMV-seropositive healthy blood donors. Successive processes, including antigen stimulation, anti-IFN-γ labeling, magnetic enrichment and elution, were then performed automatically using the CliniMACS Prodigy, which took approximately 13 h. RESULTS: The original apheresis samples consisted mainly of CD45RA+ CD62L+ naïve T cells as well as 0.3% IFN-γ-secreting CD3+ T cells that showed a response to the CMV pp65 antigen (CD3+ IFN-γ+ cells). Following IFN-γ enrichment, the target fraction contained 51.3% CD3+ IFN-γ+ cells with a reduction in naïve T cells and selection of CD45RA- CD62L- and CD45RA+ CD62L- memory T cells. Furthermore, extended culture of these isolated cells revealed functional activity, including efficient proliferation, sustained antigen-specific IFN-γ secretion, and cytotoxicity against pp65-pulsed target cells. CONCLUSION: The findings reported here suggest that the IFN-γ CCS by the CliniMACS Prodigy is a simple and robust approach to produce CMV-CTLs, which may be applicable for the treatment of clinically urgent CMV-related diseases.
Authors: Karl S Peggs; Stephanie Verfuerth; Arnold Pizzey; Naeem Khan; Malcolm Guiver; Paul A Moss; Stephen Mackinnon Journal: Lancet Date: 2003-10-25 Impact factor: 79.321
Authors: Tobias Feuchtinger; Kathrin Opherk; Wolfgang A Bethge; Max S Topp; Friedhelm R Schuster; Eva M Weissinger; Mohamad Mohty; Reuven Or; Michael Maschan; Michael Schumm; Klaus Hamprecht; Rupert Handgretinger; Peter Lang; Hermann Einsele Journal: Blood Date: 2010-07-12 Impact factor: 22.113
Authors: Georg Rauser; Hermann Einsele; Christian Sinzger; Dorothee Wernet; Gabriele Kuntz; Mario Assenmacher; John D M Campbell; Max S Topp Journal: Blood Date: 2003-12-11 Impact factor: 22.113
Authors: Carolina Berger; Michael C Jensen; Peter M Lansdorp; Mike Gough; Carole Elliott; Stanley R Riddell Journal: J Clin Invest Date: 2008-01 Impact factor: 14.808
Authors: Estéfani García-Ríos; Alejandra Leivas; Francisco J Mancebo; Laura Sánchez-Vega; Diego Lanzarot; José María Aguado; Joaquín Martínez-López; María Liz Paciello; Pilar Pérez-Romero Journal: Biomedicines Date: 2022-03-09