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The 11 th International Workshop on KSHV & Related Agents, Birmingham, UK Gene Expression I Abstract 30 HOST CELL AND VIRAL MICRORNA (MIRNA) EXPRESSION IN B CELL LYMPHOMAS Eve M Coulter, Dan Frampton, Ed Tsao, Paul Kellam MRC Centre for Medical Molecular Virology, Department of Infection, University College London, 46 Cleveland Street, London, England W1T 4JF Abstract Micro RNAs (miRNAs) are a class of approximately 21- nucleotide non-coding small RNAs that post-transcriptionally down-regulate the expression of mRNAs bearing complementary target sequences. Several hundred miRNAs have been predicted within the human genome and their key role as regulators of important biological pathways such as development, proliferation, apoptosis and viral infection is becoming clear. Recent discovery of virus-encoded miRNAs, especially miRNAs encoded by herpesviruses, indicates that viruses also use this fundamental mode of gene regulation. Using miRNA microarray technology, (470 human and 64 human viral miRNAs represented), we have determined the pattern of expression of human, Epstein Barr virus (EBV) and Kaposi’s sarcoma virus (KSHV) encoded miRNAs in B cell tumour cell lines associated with different stages of normal B cell development. Here we show that only a small proportion of human miRNAs are expressed in different B cell tumours cell lines and that difference in viral miRNA expression are evident between different herpesvirus positive tumour types. miR-155 and other human miRNAs are differentially expressed across different Bcell lymphomas including PEL. This suggests potential functional roles for their target genes in PEL biology. These data show that like virus and host cell gene expression, virus and host cell miRNA expression is associated with the underlying cell type. Presenting author Email: rebmeco@ucl.ac.uk 54
The 11 th International Workshop on KSHV & Related Agents, Birmingham, UK Gene Expression I Abstract 31 CELLULAR AND VIRAL GENE EXPRESSION PROFILING IN KSHV-INFECTED MONOCYTES Sean Gregory, Ling Wang, John West, Chelsey Hilscher, Dirk P. Dittmer and Blossom Damania Department of Microbiology and Immunology and Lineberger Cancer Center, University of North Carolina at Chapel Hill, North Carolina 27599. Abstract Kaposi's sarcoma (KS)-associated herpesvirus is the etiologic agent of Kaposi's sarcoma, primary effusion lymphoma and multicentric Castleman's disease. KSHV can infect a number of cell types including B cells, endothelial cells, epithelial cells and monocytes, macrophages and dendritic cells. We report the establishment of a monocytic cell line that is latently infected with KSHV (KSHV-THP-1). To study the host response to KSHV infection, we compared cellular gene expression in KSHV-THP-1 cells to uninfected THP-1 cells. We found that approximately six percent of cellular genes demonstrated a greater than two fold increase or decrease in gene expression in the KSHV-THP-1 cells compared to uninfected controls. Differences in gene expression of a select panel of genes were also confirmed by quantitative RT-PCR. We found that several genes involved in the host immune response were selectively downregulated in the KSHV-infected monocytes. These included tumor necrosis factor (TNF), interleukin-1b, CXCL10 and CCL4. Interestingly, the activation markers, CD86 and CD83 were also downregulated in KSHVinfected monocytes. Additionally we found several factors involved in gene expression and signal transduction to be upregulated in the KSHV-THP1 cells compared to the uninfected cells. We also profiled eighty-four viral genes in the KSHV-THP-1 cells, and found that KSHV predominantly exhibited a latent gene profile. Latency associated genes such as LANA, v-Cyclin and v-FLIP were expressed in the KSHV-infected monocytes. Presenting author Email: sgregory@med.unc.edu 55
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