title: Production of myeloid dendritic cells (DC) pulsed with tumor-specific idiotype protein for vaccination of patients with multiple myeloma.
abstract: BACKGROUND
  Immunotherapy of cancer with DC vaccines has produced encouraging results in clinical trials. Antigen (Ag)-pulsed DC have elicited CD4+ and CD8+ T-cell immunity and tumor regression in humans. However, there is no standard method of DC production. The DC phenotype, number and Ag-loading process used in these studies have varied, making comparisons between trials difficult.
  
  
  METHODS
  In the present report a reproducible method was developed for the production of a DC-based vaccine. Monocytes were enriched by adhesion from healthy donor apheresis products and cultured with growth factors for maturation into DC. The cells were loaded with the tumor Ag idiotype proteins from patients with multiple myeloma. DC culture and Ag loading were performed in an automated and closed system. The DC product was characterized for phenotype by flow cytometry and for function in Ag uptake and Ag presentation.
  
  
  RESULTS
  These monocyte-derived DC expressed high levels of costimulatory molecules (CD80/86). Ag-pulsed DC functioned to induce allogeneic proliferative lymphocyte responses and Ag-specific cytotoxic T lymphocyte (CTL) responses. The DC viability, phenotype and function were well preserved following prolonged frozen storage. Aliquots from the product of a single DC preparation could be used for sequential vaccinations without batch to batch variability.
  
  
  CONCLUSIONS
  Ag-pulsed DC can be reproducibly generated for clinical use. These standardized methods are now being employed for a clinical trial to evaluate idiotype-pulsed DC vaccine therapy following non-myeloablative transplant for the treatment of multiple myeloma.
authors.affiliation: Division of Oncology, Department of Medicine, University Hospital and Clinical Cancer Center, Stanford University Medical Center, Stanford, CA 94305, USA. guardino@stanford.edu
authors.forename: A E
authors.initials: AE
authors.lastname: Guardino
authors.forename: R
authors.initials: R
authors.lastname: Rajapaksa
authors.forename: K H
authors.initials: KH
authors.lastname: Ong
authors.forename: K
authors.initials: K
authors.lastname: Sheehan
authors.forename: R
authors.initials: R
authors.lastname: Levy
mesh_terms: D017951:Antigen Presentation / Q000276:immunology; D015703:Antigens, CD / Q000032:analysis; D015550:Cell Degranulation / Q000276:immunology; D002469:Cell Separation / Q000379:methods; D002470:Cell Survival; D018920:Coculture Techniques; D003587:Cytomegalovirus / Q000276:immunology; D003602:Cytotoxicity, Immunologic / Q000276:immunology; D003713:Dendritic Cells / Q000737:chemistry / Q000166:cytology / Q000276:immunology*; D006433:Hemocyanins / Q000737:chemistry; D006801:Humans; D007130:Immunoglobulin Idiotypes / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D016233:Immunotherapy, Active / Q000379:methods*; D007371:Interferon-gamma / Q000378:metabolism; D007937:Leukapheresis; D007959:Lymphocyte Culture Test, Mixed; D009000:Monocytes / Q000737:chemistry / Q000166:cytology / Q000276:immunology; D009101:Multiple Myeloma / Q000276:immunology / Q000628:therapy*; D022423:Myeloid Cells / Q000737:chemistry / Q000166:cytology / Q000276:immunology; D010750:Phosphoproteins / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology / Q000378:metabolism / Q000502:physiology; D014184:Transplantation, Homologous; D014763:Viral Matrix Proteins / Q000276:immunology
chemical_list: D015703:Antigens, CD; D007130:Immunoglobulin Idiotypes; D010750:Phosphoproteins; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa; D007371:Interferon-gamma; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin