title: Manufacturing of highly functional and specific T cells for adoptive immunotherapy against virus from granulocyte colony-stimulating factor-mobilized donors.
abstract: OBJECTIVE
  Cytomegalovirus (CMV) reactivation remains an important risk after hematopoietic stem cell transplantation, which can be effectively controlled through adoptive transfer of donor-derived CMV-specific T cells (CMV-T). CMV-T are usually obtained from donor peripheral blood mononuclear cells (PBMCs) collected before G-CSF mobilization. Despite previous studies that showed impaired T-cell function after granulocyte colony-stimulating factor (G-CSF) mobilization, recent publications suggest that G-CSF-primed PBMCs retain anti-viral function and are a suitable starting material for CMV-T manufacturing. The objective of this study was to assess the feasibility of generating CMV-T from G-CSF-mobilized donors by use of the activation marker CD137 in comparison with conventional non-primed PBMCs.
  
  
  METHODS
  CMV-T were isolated from G-CSF-mobilized and non-mobilized donor PBMCs on the basis of CMVpp65 activation-induced CD137 expression and expanded during 3 weeks. Functional assays were performed to assess antigen-specific activation, cytokine release, cytotoxic activity and proliferation after anti-genic re-stimulation.
  
  
  RESULTS
  We successfully manufactured highly specific, functional and cytotoxic CMV-T from G-CSF-mobilized donor PBMCs. Their anti-viral function was equivalent to non-mobilized CMV-T, and memory phenotype would suggest their long-term maintenance after adoptive transfer.
  
  
  CONCLUSIONS
  We confirm that the use of an aliquot from G-CSF-mobilized donor samples is suitable for the manufacturing of CMV cellular therapies and thereby abrogates the need for successive donations and ensures the availability for patients with unrelated donors.
authors: Oncohematology Research Group, Navarrabiomed, Miguel Servet Foundation, Pamplona, Spain.
authors: Lorea
authors: L
authors: Beloki
authors: Oncohematology Research Group, Navarrabiomed, Miguel Servet Foundation, Pamplona, Spain.
authors: Natalia
authors: N
authors: Ramírez
authors: Oncohematology Research Group, Navarrabiomed, Miguel Servet Foundation, Pamplona, Spain; Department of Haematology, Complejo Hospitalario de Navarra, Navarra Health Service, Pamplona, Spain.
authors: Eduardo
authors: E
authors: Olavarría
authors: Department of Haematology, University College London Medical School, University College London, London, United Kingdom.
authors: Edward R
authors: ER
authors: Samuel
authors: Department of Haematology, University College London Medical School, University College London, London, United Kingdom. Electronic address: m.lowdell@ucl.ac.uk.
authors: Mark W
authors: MW
authors: Lowdell
mesh_terms: D000328:Adult; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000502:physiology*; D003586:Cytomegalovirus Infections / Q000276:immunology / Q000628:therapy*; D005240:Feasibility Studies; D005260:Female; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology*; D018380:Hematopoietic Stem Cell Transplantation / Q000009:adverse effects*; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D007937:Leukapheresis / Q000379:methods*; D007963:Leukocytes, Mononuclear / Q000187:drug effects; D008297:Male; D008875:Middle Aged; D037182:T-Cell Antigen Receptor Specificity; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*; D014019:Tissue Donors; D014775:Virus Activation
chemical_list: D016179:Granulocyte Colony-Stimulating Factor
keywords: CD137; allogeneic hematopoietic stem cell transplantation; cytomegalovirus-specific T cells; granulocyte colony-stimulating factor; immunotherapy