BACKGROUND AIMS: Adoptive transfer of cytomegalovirus (CMV)-specific memory T cells can be used for treatment of CMV reactivation after allogeneic stem cell transplantation. As co-ordinated CD8(+) and CD4(+) T cells specific for a broad repertoire of CMV epitopes may be most effective for adoptive immunotherapy, the aim of this study was to isolate these cells from peripheral blood of CMV seropositive donors, irrespective of their HLA type. METHODS: Activation of CMV-specific CD8(+) and CD4(+) T cells was compared after stimulation of donor peripheral blood with minimal epitope peptides, pools of overlapping 15-mer peptides or full-length protein. Furthermore, the kinetics of interferon (IFN)-γ production after stimulation was analyzed to determine the optimal time-point for IFN-γ-based isolation of CMV-specific T cells. The specificity, phenotype and functionality of generated T-cell lines were analyzed. RESULTS: CMV protein-spanning 15-mer peptide pools induced simultaneous activation of both CD8(+) and CD4(+) CMV-specific T cells, while full-length CMV protein only efficiently activated CD4(+) CMV-specific T cells. Isolation of IFN-γ-secreting cells at the peak of the IFN-γ response after 4-h stimulation with CMV pp65 and IE1 peptide pools resulted in efficient enrichment of CMV-specific T cells. The T-cell lines contained high frequencies of CD8(+) and CD4(+) T cells recognizing multiple CMV pp65 and IE1 epitopes, and produced IFN-γ and tumor necrosis factor (TNF)-α upon specific restimulation. CONCLUSIONS: This study provides a feasible strategy for the rapid generation of clinical-grade CD8(+) and CD4(+) T-cell lines with high specificity for multiple CMV pp65 and IE1 epitopes, which may be used for effective adoptive immunotherapy.
BACKGROUND AIMS: Adoptive transfer of cytomegalovirus (CMV)-specific memory T cells can be used for treatment of CMV reactivation after allogeneic stem cell transplantation. As co-ordinated CD8(+) and CD4(+) T cells specific for a broad repertoire of CMV epitopes may be most effective for adoptive immunotherapy, the aim of this study was to isolate these cells from peripheral blood of CMV seropositive donors, irrespective of their HLA type. METHODS: Activation of CMV-specific CD8(+) and CD4(+) T cells was compared after stimulation of donor peripheral blood with minimal epitope peptides, pools of overlapping 15-mer peptides or full-length protein. Furthermore, the kinetics of interferon (IFN)-γ production after stimulation was analyzed to determine the optimal time-point for IFN-γ-based isolation of CMV-specific T cells. The specificity, phenotype and functionality of generated T-cell lines were analyzed. RESULTS: CMV protein-spanning 15-mer peptide pools induced simultaneous activation of both CD8(+) and CD4(+) CMV-specific T cells, while full-length CMV protein only efficiently activated CD4(+) CMV-specific T cells. Isolation of IFN-γ-secreting cells at the peak of the IFN-γ response after 4-h stimulation with CMV pp65 and IE1 peptide pools resulted in efficient enrichment of CMV-specific T cells. The T-cell lines contained high frequencies of CD8(+) and CD4(+) T cells recognizing multiple CMV pp65 and IE1 epitopes, and produced IFN-γ and tumor necrosis factor (TNF)-α upon specific restimulation. CONCLUSIONS: This study provides a feasible strategy for the rapid generation of clinical-grade CD8(+) and CD4(+) T-cell lines with high specificity for multiple CMV pp65 and IE1 epitopes, which may be used for effective adoptive immunotherapy.
Authors: Maarten L Zandvliet; J H Frederik Falkenburg; Ellis van Liempt; Louise A Veltrop-Duits; Arjan C Lankester; Jayant S Kalpoe; Michel G D Kester; Dirk M van der Steen; Maarten J van Tol; Roel Willemze; Henk-Jan Guchelaar; Marco W Schilham; Pauline Meij Journal: Haematologica Date: 2010-06-18 Impact factor: 9.941
Authors: Patrick J Hanley; Donald R Shaffer; Conrad R Y Cruz; Stephanie Ku; Benjamin Tzou; Hao Liu; Gail Demmler-Harrison; Helen E Heslop; Clio M Rooney; Stephen Gottschalk; Catherine M Bollard Journal: Cytotherapy Date: 2011-05-04 Impact factor: 5.414
Authors: Inge Jedema; Marian van de Meent; Jeanette Pots; Michel G D Kester; Martha T van der Beek; J H Frederik Falkenburg Journal: Haematologica Date: 2011-05-05 Impact factor: 9.941
Authors: Mattias Bronge; Klara Asplund Högelin; Olivia G Thomas; Sabrina Ruhrmann; Claudia Carvalho-Queiroz; Ola B Nilsson; Andreas Kaiser; Manuel Zeitelhofer; Erik Holmgren; Mathias Linnerbauer; Milena Z Adzemovic; Cecilia Hellström; Ivan Jelcic; Hao Liu; Peter Nilsson; Jan Hillert; Lou Brundin; Katharina Fink; Ingrid Kockum; Katarina Tengvall; Roland Martin; Hanna Tegel; Torbjörn Gräslund; Faiez Al Nimer; André Ortlieb Guerreiro-Cacais; Mohsen Khademi; Guro Gafvelin; Tomas Olsson; Hans Grönlund Journal: Sci Adv Date: 2022-04-27 Impact factor: 14.957