title: Ex vivo recovery and activation of dysfunctional, anergic, monocyte-derived dendritic cells from patients with operable breast cancer: critical role of IFN-alpha.
abstract: BACKGROUND
  Dendritic cells (DCs) play a crucial role in initiating effective cell-mediated immune responses, but are dysfunctional and anergic in breast cancer. Reversal of this dysfunction and establishment of optimal DC function is a key prerequisite for the induction of effective anti-cancer immune responses.
  
  
  RESULTS
  Peripheral blood DCs (PBDCs) and lymph node DCs (LNDCs) generated in vitro from adherent cultures of peripheral blood monocytes (PBMs) and lymph node monocytes (LNMs), respectively, using the 4 cytokine conditioned medium (CCM) (GM-CSF+IL-4+TNF-alpha+IFN-alpha) or 3 CCM (GM-CSF+IL-4+TNF-alpha) demonstrated a significantly higher degree of recovery and functional capacity in a mixed lymphocyte DC reaction (MLDCR, p < 0.001), expressed significantly higher levels of HLA-DR, CD86, compared with 2 CCM (GM-CSF+IL-4) or medium alone generated DCs from PBMs and LNMs (p < 0.001). The PBDCs generated with 3 CCM or 4 CCM showed a significantly (p < 0.001) enhanced macropinocytotic capability (dextran particles) and induced increased production and secretion of interleukin-12p40 (IL-12p40) in vitro (p < 0.001), compared with PBDCs generated from monocytes using 2 CCM or medium alone. Lipopolysaccharide (LPS) stimulation of PBDCs generated with 4 CCM demonstrated enhanced secretion of IL-6 but not IL-12p70, compared with control DCs unstimulated with LPS (p < 0.001).
  
  
  CONCLUSIONS
  Dysfunctional and anergic PBDCs and LNDCs from patients with operable breast cancer can be optimally reversed by ex vivo culturing of precursor adherent monocytes using a 4 CCM containing IFN-alpha. Maximal immunophenotypic recovery and functional reactivation of DCs is seen in the presence of IFN-alpha. However, 4 CCM containing IFN-alpha generated-PBDCs, do not produce and secrete IL-12p70 in vitro.
authors.affiliation: Section of Surgery, Queen's Medical Centre, Nottingham University Hospitals, Nottingham, UK. msxss@yahoo.co.uk
authors.forename: Sukchai
authors.initials: S
authors.lastname: Satthaporn
authors.forename: Mark M
authors.initials: MM
authors.lastname: Aloysius
authors.forename: Richard A
authors.initials: RA
authors.lastname: Robins
authors.forename: Chandan
authors.initials: C
authors.lastname: Verma
authors.forename: Suebwong
authors.initials: S
authors.lastname: Chuthapisith
authors.forename: Alasdair J
authors.initials: AJ
authors.lastname: McKechnie
authors.forename: Mohamad
authors.initials: M
authors.lastname: El-Sheemy
authors.forename: Wichai
authors.initials: W
authors.lastname: Vassanasiri
authors.forename: David
authors.initials: D
authors.lastname: Valerio
authors.forename: David
authors.initials: D
authors.lastname: Clark
authors.forename: Jibril A
authors.initials: JA
authors.lastname: Jibril
authors.forename: Oleg
authors.initials: O
authors.lastname: Eremin
mesh_terms: D051940:B7-2 Antigen / Q000276:immunology / Q000378:metabolism*; D001943:Breast Neoplasms / Q000276:immunology* / Q000378:metabolism; D002478:Cells, Cultured; D017077:Culture Media, Conditioned; D016207:Cytokines / Q000276:immunology / Q000378:metabolism*; D003713:Dendritic Cells / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D016130:Immunophenotyping; D016898:Interferon-alpha / Q000276:immunology / Q000378:metabolism*; D037181:Lectins, C-Type / Q000378:metabolism; D008214:Lymphocytes / Q000276:immunology; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins / Q000378:metabolism; D010873:Pinocytosis; D011956:Receptors, Cell Surface / Q000378:metabolism
chemical_list: D051940:B7-2 Antigen; D017077:Culture Media, Conditioned; D016207:Cytokines; D016898:Interferon-alpha; D037181:Lectins, C-Type; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins; D011956:Receptors, Cell Surface