title: A novel culture environment for generating mature human dentritic cells from peripheral blood CD14+ cells.
abstract: BACKGROUND
  We recently demonstrated that supernatants from cultures of peripheral blood mononuclear cells (PBMC) activated with anti-CD3-specific antibody (ACD3S) can induce, upon brief exposure, tumor-reactive lymphocytes in cancer patients. Here, we report that ACD3S can also induce rapid and stable maturation of dendritic cells (DC) which can be used as antigen presenting cells in in vitro protocols and for cancer immunotherapy in vivo.
  
  
  METHODS
  A short (4-day) priming of CD14+ monocytes with granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) followed by only a 24 hour-incubation in ACD3S, is sufficient to generate fully mature and stable DC.
  
  
  RESULTS
  These DC (i) stimulated strong T cell proliferative responses in the mixed lymphocyte reaction, (ii) when pulsed with unfractionated peptides from autologous tumor membrane extracts activated CD4+ T cells which proliferated in response to the autologous tumor and CD8+ cytotoxic T cells (CTL) which specifically lyse autologous tumor targets and (iii) produced high levels of IL-12.
  
  
  CONCLUSIONS
  ACD3S-treated DC are functionally superior to monocyte-conditioned medium (MCM)-treated DC generated under the same short-term protocol and as efficient as DC induced by the standard 10-day protocol. Our data present an efficient and effective method for generating in a very short period of time, highly mature and functionally competent DC.
authors.affiliation: Cancer Immunology and Immunotherapy Centre, Saint Savvas Cancer Hospital, 171 Alexandras Ave, 11522 Athens, Greece.
authors.forename: M L
authors.initials: ML
authors.lastname: Tsiatas
authors.forename: A D
authors.initials: AD
authors.lastname: Gritzapis
authors.forename: N T
authors.initials: NT
authors.lastname: Cacoullos
authors.forename: S I
authors.initials: SI
authors.lastname: Papadhimitriou
authors.forename: C N
authors.initials: CN
authors.lastname: Baxevanis
authors.forename: M
authors.initials: M
authors.lastname: Papamichail
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D001943:Breast Neoplasms / Q000473:pathology; D017252:CD3 Complex / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000166:cytology*; D005260:Female; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D016130:Immunophenotyping; D007963:Leukocytes, Mononuclear / Q000166:cytology; D018950:Lipopolysaccharide Receptors; D008175:Lung Neoplasms / Q000473:pathology; D008297:Male; D008875:Middle Aged; D010051:Ovarian Neoplasms / Q000473:pathology; D013602:T-Lymphocytes, Cytotoxic
chemical_list: D017252:CD3 Complex; D018950:Lipopolysaccharide Receptors