Project description:TLR4 signaling was reported to be involved in the upregulation of PD-L1 expression and induction of immunosuppressive properties of human CD14+ monocytes. Damage associated molecular patterns (DAMPs) S100A8, A9, and HMGB1, which act as endogenous TLR4 ligands, drive MDSC activation and were shown to be markedly expressed in the tumor microenvironment (TME) of solid tumors. The role of S100A8/9 and HMGB1 in the acquisition of MDSC immunosuppressive activity remains to be better defined.
Project description:In this study gene expression of human blood classical monocytes (CD14++CD16-), CD16 positive monocytes (consisting of non-classical CD14+16++ and intermediate CD14++CD16+ monocytes) and CD1c+ CD19- dendritic cells from healthy subjects were investigated. Keywords: expression profiling by array
Project description:The human dendritic cell (DC) family has recently been expanded by CD1c+CD14+CD163+ DCs, introduced as DC3. DC3 are found in tumors and peripheral blood of cancer patients but which cells can serve as their pre-cursors remain unknown. CD1c+CD14+ cells share similarities with both CD1c+ DCs (cDC2s) and CD14+ monocytes on transcriptomic and phenotypic level. To investigate whether CD14+ cDC2s are closer related to CD14- cDC2s or monocytes on transcriptomic level, we analyzed their RNA.
Project description:Serum-free Fibrocytes, Serum-containing Fibrocytes, CD14++CD16- Monocytes, CD14++CD16+ Monocytes, CD14+CD16++ Monocytes, Macrophages were all generated from up to 3 biological replicates from each of 3 separate donors. RNA was extracted (Ambion RNAqueous), labelled with cy3, mixed with cy5 labelled human reference (Stratagene), and hybridised to slides printed with Human AROS v4.0 oligonucleotides (Operon). Slides were scanned using a Perkin Elmer GX plus, and the data then normalised with GEPAS v4.0 and collated. Final data analysis was carried out using TMEV 4.0. SAM was performed using a 0.1% FDR. PCA were plotted from this list, and interrogation carried out using DAVID to determine pathway enrichment.
Project description:Monocytes are a heterogeneous cell population with subset-specific functions and phenotypes. The differential expression of CD14 and CD16 distinguishes classical CD14++CD16-, intermediate CD14++CD16+ and non-classical CD14+CD16++ monocytes. However, CD14++CD16+ monocytes remain the most poorly characterized subset so far. Therefore we analyzed the transcriptomes of the three monocyte subsets using SuperSAGE in combination with high-throughput sequencing. Analysis of 5,487,603 tags revealed unique identifiers of CD14++CD16+ monocytes, delineating these cells from the two other monocyte subsets. CD14++CD16+ monocytes were linked to antigen processing and presentation (e.g. CD74, HLA-DR, IFI30, CTSB), to inflammation and monocyte activation (e.g. TGFB1, AIF1, PTPN6), and to angiogenesis (e.g. TIE2, CD105). Therefore we provide genetic evidence for a distinct role of CD14++CD16+ monocytes in human immunity. Human monocyte subsets (CD14++CD16-, CD14++CD16+, CD14+CD16++) were isolated from 12 healthy volunteers based on MACS technology. Total RNA from monocyte subsets was isolated and same aliquots from each donor and monocyte subset were matched for SuperSAGE. Three SuperSAGE libraries (CD14++CD16-, CD14++CD16+ and CD14+CD16++) were generated.
Project description:Monocytes are a heterogeneous cell population with subset-specific functions and phenotypes. The differential expression of CD14 and CD16 distinguishes classical CD14++CD16-, intermediate CD14++CD16+ and non-classical CD14+CD16++ monocytes. However, CD14++CD16+ monocytes remain the most poorly characterized subset so far. Therefore we analyzed the transcriptomes of the three monocyte subsets using SuperSAGE in combination with high-throughput sequencing. Analysis of 5,487,603 tags revealed unique identifiers of CD14++CD16+ monocytes, delineating these cells from the two other monocyte subsets. CD14++CD16+ monocytes were linked to antigen processing and presentation (e.g. CD74, HLA-DR, IFI30, CTSB), to inflammation and monocyte activation (e.g. TGFB1, AIF1, PTPN6), and to angiogenesis (e.g. TIE2, CD105). Therefore we provide genetic evidence for a distinct role of CD14++CD16+ monocytes in human immunity.
Project description:Human and mouse monocytes can be divided into 2 different sub-populations, using CD14-CD16 and Ly6C-CX3CR1 respectively. We investigated the pig monocytes sub-populations and found that all porcine monocyte express CD16 and CD172M-NM-1 but can be divided into 2 subpopulation using CD14 and the scavenger receptor CD163. The CD14hi-CD163low population resemble to the inflammatory monocytes whereas the CD14low-CD163hi display more a resident monocyte type. Pig monocyte can be differentiated into macrophages when cultured with rhCSF-1 and show an increase in size, granularity and autofluorescence, and express the common macrophage markers CD14, CD16 and CD172M-NM-1. Gene expression in these 2 sub-populations was profiled using the newly-developed and annotated pig whole genome snowball microarray, showing a distinct pattern between inflammatory and resident monocytes but this difference would be more a maturation process instead of two separate subsets. Furthermore, the expression of certain genes such as CD36, CLEC4E or TREM-1 proved to share the same pattern as human monocytes, quite different from mouse monocytes. These results emphasize the potential role of the pigs as a model for human inflammatory disease and will improved our knowledge on the mononuclear phagocyte system development. Porcine PBMCs were isolated from the blood of three seperate pigs, FACS sorted on expression of CD14 and CD163 and RNA isolated from each sample, a total of 6 microarrays were hybridised
Project description:Human and mouse monocytes can be divided into 2 different sub-populations, using CD14-CD16 and Ly6C-CX3CR1 respectively. We investigated the pig monocytes sub-populations and found that all porcine monocyte express CD16 and CD172α but can be divided into 2 subpopulation using CD14 and the scavenger receptor CD163. The CD14hi-CD163low population resemble to the inflammatory monocytes whereas the CD14low-CD163hi display more a resident monocyte type. Pig monocyte can be differentiated into macrophages when cultured with rhCSF-1 and show an increase in size, granularity and autofluorescence, and express the common macrophage markers CD14, CD16 and CD172α. Gene expression in these 2 sub-populations was profiled using the newly-developed and annotated pig whole genome snowball microarray, showing a distinct pattern between inflammatory and resident monocytes but this difference would be more a maturation process instead of two separate subsets. Furthermore, the expression of certain genes such as CD36, CLEC4E or TREM-1 proved to share the same pattern as human monocytes, quite different from mouse monocytes. These results emphasize the potential role of the pigs as a model for human inflammatory disease and will improved our knowledge on the mononuclear phagocyte system development.