Unknown,Transcriptomics,Genomics,Proteomics

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Functional phenotypes alteration on dendritic cells between tolerance and immnunity

ABSTRACT: Dendritic cells (DC) are key regulators of immunological responses, immunity and tolerance. Due to their differentiation capacities, they have emerged as a promising tool for therapeutic applications. However, there are poorly understood functional DC-specific markers and cellular mechanisms for in-vitro research and clinical usage. This studyM-bM-^@M-^Ys aim is to evaluate the gene expression profiling of up-regulated or down-regulated by DC maturation and explore their function. The dendritic cells were prepared using GM-CSF and IL-4 (imDC) and subsequently maturated by adding type II collagen as antigen with LPS (mDC) or TNF-alpha(tDC). Also, surface markers, cytokine profiles, and induction of Treg cell were examined in same DC. Then, transcriptional profile was analyzed by using cDNA microarray and gene clustering programs. The selected genes, marker candidates were validated by using quantitative real-time PCR. By supervised selection, we identified 75 signitures , which 63 genes were consistently upregulated in mDC and 12 genes were expressed only in tDC . In addition, we introduced 10 genes overexpressed or equally expressed in both tDC and mDC. These selected genes were clustered into various functional categories according to GO ontology analysis. Hspa1 and Scin as tDC-specific genes, Orm1, Pdlim4, and Enpp2 as mDC-specific genes were found correlation between microarray data and mRNA expression by qRT-PCR. This work may contribute to identifying specific modulating markers for mDC and tDC, which could in the future serve as therapeutic targets in the treatment of cancer and autoimmune diseases. Total RNA obtained from isolate Bone-marrow derived Dendritic cell subset, immature DC, semi-mature or tolerogenic DC, and fully mature DC were generated in-vitro, used microarray Chip : Illumina MouseRef-8 v2 Expression BeadChip

ORGANISM(S): Mus musculus

SUBMITTER: Dae-seog Lim

PROVIDER: E-GEOD-59754 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:BACKGROUND: Combination antiretroviral therapy (cART) is able to control HIV-1 viral replication, however long-lived latent infection in resting memory CD4+ T-cells persist. The mechanisms for establishment and maintenance of latent infection in resting memory CD4+ T-cells remain unclear. Previously we have shown that HIV-1 infection of resting CD4+ T-cells co-cultured with CD11c+ myeloid dendritic cells (mDC) produced a population of non-proliferating T-cells with latent infection. Here we asked whether different antigen presenting cells (APC), including subpopulations of DC and monocytes, were able to induce post-integration latent infection in resting CD4+ T-cells, and examined potential cell interactions that may be involved using RNA-seq. RESULTS: mDC (CD1c+), SLAN+ DC and CD14+ monocytes were most efficient in stimulating proliferation of CD4+ T-cells during syngeneic culture and in generating post-integration latent infection in non-proliferating CD4+ T-cells following HIV-1 infection of APC-T-cell co-cultures. In comparison, plasmacytoid DC (pDC) and B-cells did not induce latent infection in APC-T-cell co-cultures. We compared the RNA expression profiles of APC subpopulations that could and could not induce latency in non-proliferating CD4+ T-cells. Gene expression analysis, comparing the mDC, SLAN+ DC and CD14+ monocyte subpopulations to pDC identified 53 upregulated genes that encode proteins expressed on the plasma membrane that could signal to CD4+ T-cells via cell-cell interactions (32 genes), immune checkpoints (IC) (5 genes), T-cell activation (9 genes), regulation of apoptosis (5 genes), antigen presentation (1 gene) and through unknown ligands (1 gene). CONCLUSIONS: APC subpopulations from the myeloid lineage, specifically mDC subpopulations and CD14+ monocytes, were able to efficiently induce post-integration HIV-1 latency in non-proliferating CD4+ T-cells in vitro. Inhibition of key pathways involved in mDC-T-cell interactions and HIV-1 latency may provide novel targets to eliminate HIV latency. mRNA profiles of sorted, pure antigen presenting cells including, CD1c+ myleoid dendirtic cells (mDC), SLAN+ mDC, CD14+ monocytes and plasmacytoid DC (pDC), were generated using next generation sequencing in triplicate, using Illumina Illumina Hiseq 2000.

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Functional phenotypes alteration on dendritic cells between tolerance and immnunity

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