Molecular signatures of maturing dendritic cells: implications for testing the quality of dendritic cell therapies

Abstract

BackgroundDendritic cells (DCs) are often produced by granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) stimulation of monocytes. To improve the effectiveness of DC adoptive immune cancer therapy, many different agents have been used to mature DCs. We analyzed the kinetics of DC maturation by lipopolysaccharide (LPS) and interferon-γ (IFN-γ) induction in order to characterize the usefulness of mature DCs (mDCs) for immune therapy and to identify biomarkers for assessing the quality of mDCs.MethodsPeripheral blood mononuclear cells were collected from 6 healthy subjects by apheresis, monocytes were isolated by elutriation, and immature DCs (iDCs) were produced by 3 days of culture with GM-CSF and IL-4. The iDCs were sampled after 4, 8 and 24 hours in culture with LPS and IFN-γ and were then assessed by flow cytometry, ELISA, and global gene and microRNA (miRNA) expression analysis.ResultsAfter 24 hours of LPS and IFN-γ stimulation, DC surface expression of CD80, CD83, CD86, and HLA Class II antigens were up-regulated. Th1 attractant genes such as CXCL9, CXCL10, CXCL11 and CCL5 were up-regulated during maturation but not Treg attractants such as CCL22 and CXCL12. The expression of classical mDC biomarker genes CD83, CCR7, CCL5, CCL8, SOD2, MT2A, OASL, GBP1 and HES4 were up-regulated throughout maturation while MTIB, MTIE, MTIG, MTIH, GADD45A and LAMP3 were only up-regulated late in maturation. The expression of miR-155 was up-regulated 8-fold in mDCs.ConclusionDCs, matured with LPS and IFN-γ, were characterized by increased levels of Th1 attractants as opposed to Treg attractants and may be particularly effective for adoptive immune cancer therapy.

MicroRNA Expression Profiles of Human Blood Monocyte-derived Dendritic Cells and Macrophages Reveal miR-511 as Putative Positive Regulator of Toll-like Receptor 4

“…In this study, we first describe updated miRNA expression profiles of human blood MO-derived immature and mature DCs and MFs. Consistent with previous studies (23,27,31,42,43), we found that these cell types have distinct miRNA expression profiles and detected several novel strongly up-regulated miRNAs, including miR-511. For the activation and maturation of MFs and DCs, we used LPS and curdlan as ligands for TLR4 and Dectin-1, respectively, both leading to activation through the NF-B pathway (44 -46).…”

The α7β0 Isoform of the Complement Regulator C4b-Binding Protein Induces a Semimature, Anti-Inflammatory State in Dendritic Cells

“…In contrast, the anti-inflammatory and immunoregulatory cytokine TGF-b (39) was maintained at levels approaching those from iDCs. These results are in agreement with the recently reported molecular signature of LPS/IFN-g-matured monocyte-derived DCs (24). Our ongoing work comprehensively assessing the genome-wide comparative transcriptional profile of proinflammatory DCs either untreated or in the presence of C4BP(b + ) or C4BP(b 2 ) isoforms will delineate the complete molecular signature of C4BP(b 2 )-induced tolerogenic DCs.…”

TIE-2 and VEGFR Kinase Activities Drive Immunosuppressive Function of TIE-2–Expressing Monocytes in Human Breast Tumors

“…4B). Some of these genes have been previously reported to be upregulated upon in vitro maturation of DCs following exposure to lipopolysaccharide and interferon-g (21,22). Consistent with this observation, the expression of genes related to antigen processing and presentation, which are specific functions of APCs, were also upregulated (Fig.…”