title: Detailed analysis of cytomegalovirus (CMV)-specific T cells expanded for adoptive immunotherapy of CMV infection following allogeneic stem cell transplantation for malignant disease.
abstract: BACKGROUND
  Cytomegalovirus (CMV) infection and its treatment causes significant morbidity following allogeneic stem cell transplantation (SCT) for malignancies. We studied the phenotype, function and growth kinetics of CMV pp65 antigen (Ag)-specific T cells expanded in a short-term culture for adoptive therapy.
  
  
  METHODS
  Peripheral blood mononuclear cells (PBMC) from CMV-seropositive donors were cultured in various conditions with CMV pp65((495-503)) peptide to determine the most effective method for generating CMV-specific T cells. CMV-expanded cultures were tested for frequency, phenotype and functionality using peptide-MHC tetramer analysis, cytokine flow cytometry and cytolytic assays. A patient undergoing allogeneic SCT was administered CMV pp65-specific T cells generated from the donor based on these data, and recipient PBMC were analyzed following T-cell infusion.
  
  
  RESULTS
  CMV pp65-specific T cells were consistently generated from CMV-seropositive donors at high frequencies (20-40% of CD8+ T cells), secreted interferon-gamma (IFN-gamma) in response to CMV peptide and had lytic activity against CMV peptide-expressing targets. Cultured CMV-specific T cells were infused into a SCT recipient without toxicity.
  
  
  CONCLUSIONS
  Stimulating donor PBMC to generate functional, Ag-specific T cells for infusion into SCT recipients was accomplished consistently using readily available technology. We observed no toxicity in one patient receiving T cells and were able to monitor infused cells. These findings support further study of this approach as a prophylaxis against the risk of infection in patients receiving allogeneic transplantation from CMV-seropositive donors.
authors.affiliation: Department of Surgery, Duke University Medical Center, Durham, North Carolina, USA.
authors.forename: Ac
authors.initials: A
authors.lastname: Hobeika
authors.forename: T
authors.initials: T
authors.lastname: Osada
authors.forename: D
authors.initials: D
authors.lastname: Serra
authors.forename: S
authors.initials: S
authors.lastname: Peplinski
authors.forename: K
authors.initials: K
authors.lastname: Hanson
authors.forename: Y
authors.initials: Y
authors.lastname: Tanaka
authors.forename: D
authors.initials: D
authors.lastname: Niedzwiecki
authors.forename: N
authors.initials: N
authors.lastname: Chao
authors.forename: D
authors.initials: D
authors.lastname: Rizzieri
authors.forename: Hk
authors.initials: H
authors.lastname: Lyerly
authors.forename: T
authors.initials: T
authors.lastname: Clay
authors.forename: Ma
authors.initials: M
authors.lastname: Morse
mesh_terms: D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D002999:Clone Cells; D016207:Cytokines / Q000378:metabolism; D003587:Cytomegalovirus / Q000187:drug effects / Q000276:immunology*; D003586:Cytomegalovirus Infections / Q000276:immunology*; D000939:Epitopes; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D016219:Immunotherapy, Adoptive; D007700:Kinetics; D008213:Lymphocyte Activation / Q000187:drug effects; D009369:Neoplasms / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D010455:Peptides / Q000494:pharmacology; D010641:Phenotype; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000276:immunology; D033581:Stem Cell Transplantation; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D014184:Transplantation, Homologous
chemical_list: D016207:Cytokines; D000939:Epitopes; D010455:Peptides; D016693:Receptors, Antigen, T-Cell, alpha-beta