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Title:   Author  item
                                            ZHANG, WEI - CASE WESTERN UNIVERSITYitem
                                            DING, JIANQING - RUSH UNIVERSITYitem
                                            QU, YAN - CASE WESTERN UNIVERSITYitem
                                            HU, HONGLIANG - RUSH UNIVERSITYitem
                                            LIN, MEIHUA - CASE WESTERN UNIVERSITYitem
                                            DATTA, AMIT - CASE WESTERN UNIVERSITYitem
                                            LARSON, ALAN - RUSH UNIVERSITYitem
                                            Liu, Ge

                                                         - *George*item
                                            LI, BIAORU - CASE WESTERN & RUSH UNIV
  Submitted to: Immunology
                                    Publication Type: Peer Reviewed Journal
                                    Publication Acceptance Date: 7/22/2008
                                    Publication  Date: 9/5/2008
                                    Citation: Zhang, W., Ding, J., Qu, Y., Hu, H., Lin, M., Datta, A., Larson, A., Liu, G., Li, B. 2008. Genomic expression analysis by single-cell mRNA differential display of quiescent CD8 T cells from tumor-infiltrating lymphocytes obtained from in vivo liver tumours. Immunology. 127(1):83-90.Interpretive Summary: The quiescent state of T lymphocytes was thought to be due to the lack of activation signals. However, recent studies indicated that quiescence in CD8 T cells was an actively maintained state rather than a defective state resulting from the absence of the stimulation signals. The molecular mechanisms underlying the quiescent status in CD8 T cell of TIL remains unclear due to the technological limitation of analyzing the small number of T cells within cancer tissues. Here we describe a genomic approach combining single-cell mRNA differential display and RNA subtractive hybridization to elucidate the mechanism of the CD8 T-cell ignorance. By comparing the quiescent CD8 T-cell obtained from liver tumor TIL with a reference control at the single cell level, we identified candidate genes for differential expression profile by high throughput screening and comparative analysis of expressed sequence tags (ESTs). While T-cell receptor, TNF receptor, TRAIL and perforin are down-regulated, several key genes (such as Tob, TGF-beta, LKLF, SnoA, Ski, Myc, ERF and REST/NRSF complex) are highly expressed in the quiescent CD8 cells. Tob and TGF-' expression are significantly decreased after reactivating the quiescent T cells by IL-2. Real time PCR further confirmed these expression profiles. Based on these data, a regulation model of CD8 T-cell quiescence is proposed including three pathways, which are up-regulation of TGF-beta pathway, shift of myc web and inhibition of cell cycle. These results will set up a good basis to study the mechanism of CD8 cell ignorance.
                                Technical Abstract: We performed a genomic approach combining single-cell mRNA differential display and RNA subtractive hybridization to elucidate the CD8 T-cell ignorance. By comparing the quiescent CD8 T-cell obtained from liver tumor TIL with a control at the single cell level, we identified candidate genes differentially expressed by high-throughput screening and comparative analysis of expressed sequence tags (ESTs). While T-cell receptor, TNF receptor, TRAIL and perforin are down-regulated, key genes like Tob, TGF-beta, LKLF, SnoA, Ski, Myc, ERF and REST/NRSF complex are highly expressed in the quiescent CD8 cells. Real time PCR further confirmed these results. A regulation model of CD8 T-cell quiescence is proposed including three ways: up-regulation of TGF-beta pathway, shift of myc web and inhibition of cell cycle.


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