Page Banner

United States Department of Agriculture

Agricultural Research Service

Title: Genomic expression analysis by single-cell mRNA differential display of quiescent CD8 T cells from tumor-infiltrating lymphocytes obtained from in vivo liver tumours)

Author
item Zhang, Wei
item Ding, Jianqing
item Qu, Yan
item Hu, Hongliang
item Lin, Meihua
item Datta, Amit
item Larson, Alan
item Liu, Ge - George
item Li, Biaoru

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.

Last Modified: 8/24/2016
Footer Content Back to Top of Page