Abstract
Most anti-cancer immunotherapeutic strategies involving dendritic cells (DC) as vaccines rely upon the adoptive transfer of DC loaded with exogenous tumour-peptides. This study utilized human acute myeloid leukemia (AML) cells as progenitors from which functional dendritic-like antigen presenting cells (DLC) were generated, that constitutively express tumour antigens for recognition by CD8+ T cells. DLC were generated from AML cell lines KG-1 and MUTZ-3 using rapid culture techniques and appropriate cytokines. DLC were evaluated for their cell-surface phenotype, antigen uptake and ability to stimulate allogeneic responder cell proliferation, and production of IFN-γ; compared with DC derived from normal human PBMC donors. KG-1 and MUTZ-3 DLC increased expression of CD80, CD83, CD86, and HLA-DR, and MUTZ-3 DLC downregulated CD14 and expressed CD1a. Importantly, both KG-1 and MUTZ-3-derived DLC promoted proliferation of allogeneic responder cells more efficiently than unmodified cells; neither cells incorporated FITC-labeled dextran, but both stimulated IFN-γ production from responding allogeneic CD8+ T cells. Control DC produced from PBMC using the FastDC culture also expressed high levels of critical cell surface ligands and demonstrated good APC function. This paper indicates that functional DLC can be cultured from the AML cell lines KG-1 and MUTZ-3, and FastDC culture generates functional KG-1 DLC.
Highlights
Manipulating the interactions between dendritic cells (DC) and naıve T cells is an important part of the design of strategies for the immunotherapy of both solid and hematological cancers [1]
This paper indicates that functional dendritic-like antigen presenting cells (DLC) can be cultured from the acute myeloid leukemia (AML) cell lines KG-1 and MUTZ-3, and FastDC culture generates functional KG-1 DLC
It was reported that functional DC could be cultured from peripheral blood mononuclear cells (PBMC) using a combination of GM-CSF and IL-4 for 24 hours followed by a combination of four proinflammatory cytokines (TNF-α, IL-1β, IL-6, and PGE2) for a further 24 hours [3]
Summary
Manipulating the interactions between DC and naıve T cells is an important part of the design of strategies for the immunotherapy of both solid and hematological cancers [1]. DC can be generated from CD34+ cells by culture with cytokines such as GM-CSF, IL-4, Flt3L, CD40 ligand, SCF, and TGF-β. DC can be grown from CD14+ peripheral blood mononuclear cells (PBMC) during 5–9day culture, using GM-CSF and IL-4 [2]. It was reported that functional DC could be cultured from PBMC using a combination of GM-CSF and IL-4 for 24 hours followed by a combination of four proinflammatory cytokines (TNF-α, IL-1β, IL-6, and PGE2) for a further 24 hours [3]. We and others have previously reported that DC can be cultured from leukemic blast cells [4,5,6]. In this study we have utilized AML cell lines for use as a source of DLC. This approach provides a uniform source of DLC for further study
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