Ex vivo recovery and activation of dysfunctional, anergic, monocyte-derived dendritic cells from patients with operable breast cancer: critical role of IFN-alpha.
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.
Satthaporn
Sukchai
S
Section of Surgery, Queen's Medical Centre, Nottingham University Hospitals, Nottingham, UK. msxss@yahoo.co.uk
Aloysius
Mark M
MM
Robins
Richard A
RA
Verma
Chandan
C
Chuthapisith
Suebwong
S
McKechnie
Alasdair J
AJ
El-Sheemy
Mohamad
M
Vassanasiri
Wichai
W
Valerio
David
D
Clark
David
D
Jibril
Jibril A
JA
Eremin
Oleg
O
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
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