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. 2010 Jan 15:8:4.
doi: 10.1186/1479-5876-8-4.

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

Ping Jin  1 , Tae Hee Han, Jiaqiang Ren, Stefanie Saunders, Ena Wang, Francesco M Marincola, David F Stroncek
Affiliations

"V体育2025版" Molecular signatures of maturing dendritic cells: implications for testing the quality of dendritic cell therapies

Ping Jin et al. J Transl Med. .

Abstract

Background: Dendritic 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-gamma (IFN-gamma) induction in order to characterize the usefulness of mature DCs (mDCs) for immune therapy and to identify biomarkers for assessing the quality of mDCs VSports手机版. .

Methods: Peripheral 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 V体育安卓版. The iDCs were sampled after 4, 8 and 24 hours in culture with LPS and IFN-gamma and were then assessed by flow cytometry, ELISA, and global gene and microRNA (miRNA) expression analysis. .

Results: After 24 hours of LPS and IFN-gamma 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. V体育ios版.

Conclusion: DCs, matured with LPS and IFN-gamma, were characterized by increased levels of Th1 attractants as opposed to Treg attractants and may be particularly effective for adoptive immune cancer therapy. VSports最新版本.

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Figures

Figure 1
Figure 1
Gene expression changes in maturing DCs. Immature DCs from 6 healthy subjects were incubated with LPS and IFN-γ. After 0, 4, 8, and 24 hours of culture, DCs were analyzed by gene expression profiling using a microarray with 35,035 oligonucleotide probes. The 2,370 differentially expressed genes (F-test: p < 0.001) were analyzed by supervised hierachical clustering. Immature DCs are indicated by the orange bar, iDCs cultured with LPS and IL-4 for 4 hours by the green bar, 8 hours by the purple bar, and 24 hours by the red bar. The genes sorted into 5 separate clusters and representive genes from each of the 5 clusters are shown.
Figure 2
Figure 2
Change in the expression of CCR7, CD86, HLA-DRA, and HLA-DRB during DC maturation. iDCs from 6 healthy subjects sampled after 0, 4, 8 and 24 hours of culture in LPS and IFN-γ were analyzed by quantitive RT-PCR for the expression of CCR7, CD86, HLA-DRA, and HLA-DRB.
Figure 3
Figure 3
miR expression changes in maturing DCs. Immature DCs from 6 healthy subjects were incubated with LPS and IFN-γ and after 0, 4, 8, and 24 hours of culture, they were analyzed by global microRNA expression profiling using a microarray with 827 probes. The differentially expressed human miR (F-test: p < 0.05) were analyzed by supervised hierachical clustering. The samples clustered into 4 groups based on maturation time, iDC are indicated by the green bar, DCs cultured for 4 hours by the orange bar, DCs cultured for 8 hours by the red bar, and 24 hours by the purple bar. The miRs sorted into 2 separate clusters and miRs from each of the clusters are shown.
Figure 4
Figure 4
Change in the expression of miR-146a, -146b, and -155 during DC maturation. iDCs from 6 healthy subjects sampled after 0, 4, 8 and 24 hours of culture in LPS and IFN-γ were analyzed by quantitive RT-PCR for the expression of miR-146a, -146b, and -155.
Figure 5
Figure 5
Cytokine, chemokine and growth factors production by cultured DCs. The supernants from the 6 healthy subject iDCs and mDCs from 6 healthy subjects were analyzed for 50 cytokines, chemokines and growth factors using an ELISA assay. The levels of 36 soluble factors differed between iDCs and mDCs (t-tests, p < 0.05), the levels of all 36 were greater in mDCs. The differentially expressed factors were analyzed by unsupervised hierachical clustering analysis.
Figure 6
Figure 6
Production of cytokine, chemokine and growth factors by stimulated mDCs. Mature DCs from 6 healthy subjects produced by incubation with LPS and IFN-γ were incubated with mouse fibroblasts transfected with CD40-Ligand (CD40L-LTK). The supernant from the stimulated mDCs and unstimulated mDCs were analyzed for 50 cytokines, chemokines and growth factors using an ELISA assay. The levels of the 50 soluble factors were analyzed by unsupervised hierachical clustering analysis. The DCs separated into stimulated mDCs (purple bar) and unstimulated mDCs (orange bar). The cluster of factors that were increased in stimulated mDCs are shown in the purple box and those decreased in stimulated mDCs are shown in the orange box.
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References

    1. Hashimoto SI, Suzuki T, Nagai S, Yamashita T, Toyoda N, Matsushima K. Identification of genes specifically expressed in human activated and mature dendritic cells through serial analysis of gene expression. Blood. 2000;96:2206–2214. - PubMed
    1. Young JW, Merad M, Hart DN. Dendritic cells in transplantation and immune-based therapies. Biol Blood Marrow Transplant. 2007;13:23–32. doi: 10.1016/j.bbmt.2006.10.023. - DOI - PubMed
    1. Randolph GJ, Jakubzick C, Qu C. Antigen presentation by monocytes and monocyte-derived cells. Curr Opin Immunol. 2008;20:52–60. - PMC - PubMed
    1. Banchereau J, Palucka AK. Dendritic cells as therapeutic vaccines against cancer. Nat Rev Immunol. 2005;5:296–306. doi: 10.1038/nri1592. - DOI - PubMed
    1. Gilboa E. DC-based cancer vaccines. J Clin Invest. 2007;117:1195–1203. doi: 10.1172/JCI31205. - DOI - PMC - PubMed

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