Project description:To elucidate the difference between CD161-negative TCR Vdelta1 and CD161-positive TCR Vdelta1 cells, we performed GeneChip analysis of these two types of cells in peripheral blood mononuclear cells from healthy donors. Such analysis identified 192 genes that were expressed at two-fold or higher levels on CD161-positive TCR Vdelta1 cells than on CD161-negative TCR Vdelta1 cells. Expression of three genes (CCL3, CCL4 and CD161) from this signature was quantified in the same RNA samples by real-time PCR. CD161-negative and CD161-positive TCR Vdelta1cells of three healthy volunteers were isolated from freshly peripheral blood mononuclear cells.
Project description:To elucidate the difference between CD161-negative TCR Vdelta1 and CD161-positive TCR Vdelta1 cells, we performed GeneChip analysis of these two types of cells in peripheral blood mononuclear cells from healthy donors. Such analysis identified 192 genes that were expressed at two-fold or higher levels on CD161-positive TCR Vdelta1 cells than on CD161-negative TCR Vdelta1 cells. Expression of three genes (CCL3, CCL4 and CD161) from this signature was quantified in the same RNA samples by real-time PCR.
Project description:Gene expression profiling of immortalized human mesenchymal stem cells with hTERT/E6/E7 transfected MSCs. hTERT may change gene expression in MSCs. Goal was to determine the gene expressions of immortalized MSCs.
Project description:We compared gammadelta T-cell development and subsets in the thymus of mice in which Shh had been conditionally deleted from thymic epithelial cells using FoxN1-Cre to WT mice, and to transgenic mice in which a constitutively activator (Gli2DN2) or constitutuvely repressor (Gli2DC2) form of Gli2 were expressed under the control of the lck-promotor. We sorted CD27+CD3+TCR-gammadelta+ cells from 4 week old mice and comapred gene expression by RNAsequencing. All mice were on a C57BL/6 background.
Project description:Transcriptional profiling of human mesenchymal stem cells comparing normoxic MSCs cells with hypoxic MSCs cells. Hypoxia may inhibit senescence of MSCs during expansion. Goal was to determine the effects of hypoxia on global MSCs gene expression.
Project description:Gene methylation profiling of immortalized human mesenchymal stem cells comparing HPV E6/E7-transfected MSCs cells with human telomerase reverse transcriptase (hTERT)- and HPV E6/E7-transfected MSCs. hTERT may increase gene methylation in MSCs. Goal was to determine the effects of different transfected genes on global gene methylation in MSCs.
Project description:Kynureninase is a member of a large family of catalytically diverse but structurally homologous pyridoxal 5'-phosphate (PLP) dependent enzymes known as the aspartate aminotransferase superfamily or alpha-family. The Homo sapiens and other eukaryotic constitutive kynureninases preferentially catalyze the hydrolytic cleavage of 3-hydroxy-l-kynurenine to produce 3-hydroxyanthranilate and l-alanine, while l-kynurenine is the substrate of many prokaryotic inducible kynureninases. The human enzyme was cloned with an N-terminal hexahistidine tag, expressed, and purified from a bacterial expression system using Ni metal ion affinity chromatography. Kinetic characterization of the recombinant enzyme reveals classic Michaelis-Menten behavior, with a Km of 28.3 +/- 1.9 microM and a specific activity of 1.75 micromol min-1 mg-1 for 3-hydroxy-dl-kynurenine. Crystals of recombinant kynureninase that diffracted to 2.0 A were obtained, and the atomic structure of the PLP-bound holoenzyme was determined by molecular replacement using the Pseudomonas fluorescens kynureninase structure (PDB entry 1qz9) as the phasing model. A structural superposition with the P. fluorescens kynureninase revealed that these two structures resemble the "open" and "closed" conformations of aspartate aminotransferase. The comparison illustrates the dynamic nature of these proteins' small domains and reveals a role for Arg-434 similar to its role in other AAT alpha-family members. Docking of 3-hydroxy-l-kynurenine into the human kynureninase active site suggests that Asn-333 and His-102 are involved in substrate binding and molecular discrimination between inducible and constitutive kynureninase substrates.
Project description:Gene expression profiling of immortalized human mesenchymal stem cells with hTERT/E6/E7 transfected MSCs. hTERT may change gene expression in MSCs. Goal was to determine the gene expressions of immortalized MSCs. One-condition experment, gene expression of 3A6