title: Manufacture of GMP-compliant functional adenovirus-specific T-cell therapy for treatment of post-transplant infectious complications.
abstract: In pediatric patients, adenovirus (ADV) reactivation after allogeneic hematopoietic stem cell transplantation (allo HSCT) is a major cause of morbidity and mortality. For patients who do not respond to antiviral drug therapy, a new treatment approach using ADV-specific T cells can present a promising alternative. Here we describe the clinical scale Good Manufacturing Practice (GMP)-compliant manufacture and characterization of 40 ADV-specific T-cell products, Cytovir ADV, which are currently being tested in a multi-center phase I/IIa clinical trial. This process requires minimal intervention, is high yield, and results in a pure T-cell product that is functional.
  
  
  
  Mononuclear cells (2 × 10(7)) were cultured in a closed system in the presence of GMP-grade ADV peptide pool and cytokines for 10 days. On day 10, the T-cell product was harvested, washed in a closed system, counted and assessed for purity and potency. Additional characterization was carried out where cell numbers allowed.
  
  
  
  Thirty-eight of 40 products (95%) met all release criteria. Median purity of the cell product was 88.3% CD3+ cells with a median yield of 2.9 × 10(7) CD3+ cells. Potency analyses showed a median ADV-specific interferon (IFN)γ response of 5.9% of CD3+ and 2345 IFNγ spot-forming cells/million. CD4 and CD8 T cells were capable of proliferating in response to ADV (63.3 and 56.3%, respectively). These virus-specific T cells (VST) were heterogenous, containing both effector memory and central memory T cells. In an exemplar patient with ADV viremia treated in the open ASPIRE trial, ADV-specific T-cell response was detected by IFNγ enzyme-linked immunospot from 13 days post-infusion. ADV DNA levels declined following cellular therapy and were below level of detection from day 64 post-infusion onward.
  
  
  
  The clinical-scale GMP-compliant One Touch manufacturing system is feasible and yields functional ADV-specific T cells at clinically relevant doses.
authors.affiliation: Cell Medica Ltd, London, United Kingdom. Electronic address: claire.horlock@cellmedica.co.uk.
authors.forename: Claire
authors.initials: C
authors.lastname: Horlock
authors.affiliation: Cell Medica Ltd, London, United Kingdom.
authors.forename: Amanda
authors.initials: A
authors.lastname: Skulte
authors.affiliation: Cell Medica Ltd, London, United Kingdom.
authors.forename: Arindam
authors.initials: A
authors.lastname: Mitra
authors.affiliation: Cell Medica Ltd, London, United Kingdom.
authors.forename: Alka
authors.initials: A
authors.lastname: Stansfield
authors.affiliation: Cell Medica Ltd, London, United Kingdom.
authors.forename: Shristi
authors.initials: S
authors.lastname: Bhandari
authors.affiliation: Great Ormond Street Hospital, London, United Kingdom.
authors.forename: Winnie
authors.initials: W
authors.lastname: Ip
authors.affiliation: Great Ormond Street Hospital, London, United Kingdom.
authors.forename: Waseem
authors.initials: W
authors.lastname: Qasim
authors.affiliation: Research Department of Haematology, Cancer Institute, UCL, Royal Free Hospital, London, United Kingdom.
authors.forename: Mark W
authors.initials: MW
authors.lastname: Lowdell
authors.affiliation: Cell Medica Ltd, London, United Kingdom.
authors.forename: Shreenal
authors.initials: S
authors.lastname: Patel
authors.affiliation: Cell Medica Ltd, London, United Kingdom.
authors.forename: Anke
authors.initials: A
authors.lastname: Friedetzky
authors.affiliation: Cell Medica Ltd, London, United Kingdom.
authors.forename: Marco A
authors.initials: MA
authors.lastname: Purbhoo
authors.affiliation: Cell Medica Ltd, London, United Kingdom.
authors.forename: Katy
authors.initials: K
authors.lastname: Newton
mesh_terms: D000256:Adenoviridae / Q000472:pathogenicity / Q000502:physiology; D000257:Adenoviridae Infections / Q000628:therapy; D018929:Cell Culture Techniques / Q000379:methods* / Q000592:standards; D018380:Hematopoietic Stem Cell Transplantation / Q000009:adverse effects*; D006801:Humans; D016130:Immunophenotyping; D007167:Immunotherapy / Q000379:methods; D013601:T-Lymphocytes / Q000166:cytology* / Q000821:virology
keywords: ADV; GMP compliant; HSCT; adoptive T-cell therapy