Project description:Primary diffuse large B cell lymphomas of different immune-privileged sites (IP-DLBCL) share many clinical and biological features, such as a relatively poor prognosis, preferential dissemination to other immune-privileged sites and deletion of the HLA region, which suggests that IP-DLBCL represents a separate entity. To further investigate the nature of IP-DLBCL, we investigated site-specific genomic aberrations in 16 testicular, 9 central nervous system (CNS) and 15 nodal DLBCL using array-CGH. We also determined minimal common regions of gain and loss. Using robust algorithms, the array-CGH data were combined with gene expression data to explore pathways deregulated by chromosomal aberrations. Keywords: comparative genomic hybridisation, gene expression, tumor type comparison Whole tissue sections of 16 testicular, 9 central nervous system (CNS) and 15 nodal DLBCL were used for isolation of genomic DNA and total RNA. Genomic aberrations were determined using an in house printed CGH array containing ~3700 large genomic insert clones. Gene expression was analyzed on Affymetrix HU133 plus 2.0 oligo arrays. Gene expression data and arrayCGH data were combined using the R program ACE-it.
Project description:RNAseq analysis of human lung slices after treatment with different interferons. Results: Differentially expressed genes were observed after treatment with different interferons. Methods: Tumor-free human lung explants were obtained from patients undergoing lung surgery at the University Hospital Muenster on the day of surgery. Tissue blocks were individually placed in a 12-well plate in RPMI medium. Tissues were treated with different interferons. RNA was prepared from tissue homogenates.
Project description:Primary diffuse large B cell lymphomas of different immune-privileged sites (IP-DLBCL) share many clinical and biological features, such as a relatively poor prognosis, preferential dissemination to other immune-privileged sites and deletion of the HLA region, which suggests that IP-DLBCL represents a separate entity. To further investigate the nature of IP-DLBCL, we investigated site-specific genomic aberrations in 16 testicular, 9 central nervous system (CNS) and 15 nodal DLBCL using array-CGH. We also determined minimal common regions of gain and loss. Using robust algorithms, the array-CGH data were combined with gene expression data to explore pathways deregulated by chromosomal aberrations. Keywords: comparative genomic hybridisation, gene expression, tumor type comparison
Project description:The class I major histocompatibility complex (MHC) bound peptides, produced from immature proteins that are degraded rapidly are called defective ribosome products (DRiPs). Such DRiPs are possibly involved with early alerting of the immune system about impeding infections, thus bringing about faster killing of infected cells before the viruses replicate. Interferons are a major group of cytokines, produced in response to viral infection. The interferons modulate the cellular metabolism and gene expression patterns and increase the expression and cell-surface presentation of the MHC molecules, helping this way coping with the viral infection. In this study, we evaluated whether the interferons also induce rapid degradation of cellular or viral proteins and produce DRiPs MHC-bound peptides to help the alert of the immune system. Cultured human breast cancer cells were treated with interferons and the cells were transferred to growth media containing heavy stable isotope labeled amino acids (dynamic-SILAC)in several time points a few hours after the interferons’ treatment. The rates of synthesis, degradation, and production of the cellular protein and their degradation products, the MHC peptides, were followed by LC-MS/MS analyses. Detection of large numbers of MHC peptides that incorporate the heavy amino acids into them, faster than their source proteins, indicated to us that not only DRiP-peptides are rather abundant among the MHC peptidome, but that the interferons increase significantly the presentation of DRiP-peptides. Importantly, much of this DRiPome derive from multi-subunit complexes, including the proteasomes and ribosomes, while the degradation of the standard protostome give rise to the immunoproteasome, which is thought to produce peptides that enhance the immune-response; the degradation of the ribosome subunits may aid in reducing the synthesis of viral infection
Project description:Type I Interferons encompasses a large family of closely related cytokines comprising of at least 13 IFN-α isotypes and single IFN-β. Both IFN-α and IFN-β exert their activity through a common receptor IFNAR. Type I Interferons have broad regulatory effects and various subtypes of dendritic cells are influenced by this cytokines. In our study we asked question whether the low, constitutive levels of type I Interferons produced under steady state conditions are important for proper function of splenic conventional dendritic cells. In this approach we sorted out two populations (CD8α+ and CD8α-) of splenic dendritic cells (DCs) from untreated WT, IFN-β-/- and IFNAR-/- C57Bl/6 mice. All mice were between 8-10 weeks old. Further we isolated RNA and performed microarray analysis. Each DCs population was repeated twice.