Ex vivo recovery and activation of dysfunctional, anergic, monocyte-derived dendritic cells from patients with operable breast cancer: critical role of IFN-alpha

Sukchai Satthaporn,Suebwong Chuthapisith,Chandan Verma,Wichai Vassanasiri,Alasdair J Mckechnie,Oleg Eremin,David Clark,Mark M Aloysius,Richard A Robins,David Valerio,Mohamad El-Sheemy,Jibril A Jibril

doi:10.1186/1471-2172-9-32

Abstract

BackgroundDendritic 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.ResultsPeripheral 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-α+IFN-α) or 3 CCM (GM-CSF+IL-4+TNF-α) 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).ConclusionDysfunctional 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-α. Maximal immunophenotypic recovery and functional reactivation of DCs is seen in the presence of IFN-α. However, 4 CCM containing IFN-α generated-PBDCs, do not produce and secrete IL-12p70 in vitro.

Highlights

Dendritic cells (DCs) play a crucial role in initiating effective cell-mediated immune responses, but are dysfunctional and anergic in breast cancer
We have previously shown in women with operable breast cancer, that both peripheral blood (PBDCs) and tumourdraining axillary lymph node DCs (LNDCs) were dysfunctional and switched off, as assessed by phenotypic profile (HLA-DR, CD86 and CD40) and functional assays in vitro (MLDCR, autologous purified protein derivative (PPD) stimulation assay) [15]
Peripheral blood monocytes and lymph node monocytes (LNMs) grown in various cytokine conditioned medium (CCM); 3 CCM (GM-CSF+IL-4+tumour necrosis factor-α (TNF-α)) or 4 CCM (GMCSF+IL-4+TNF-α+ IFN-α), expressed high levels of HLA class II proteins, CD86, consistent with their differentiation into activated DCs compared with DCs cultured with 2 CCM (GM-CSF + IL-4) or medium alone (Figure 1A, B and 1C)

Summary

Introduction

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. Up-regulation of class I and class II major histocompatibility (MHC) proteins, CD80 and CD86 co-stimulatory molecules, and CD40 adhesion molecule expression enhances the ability of DCs to present Ag to naïve T lymphocytes in secondary lymphoid compartments and induce their activation and generation of peptide-specific cytotoxic T lymphocytes (CTLs) [5]. Activated and mature DCs produce interleukin-12 (IL-12), which stimulates CD4+ T helper 1 (Th1) cell activation and development, which is important for the in situ (paracortex of lymph nodes) expansion of induced anti-tumour CTLs [68]

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