Depletion of Cytoplasmic Tail of UL18 Enhances and Stabilizes the Surface Expression of UL18

BACKGROUND: Human cytomegalovirus UL18, a MHC class I homologue, has been considered a natural killer (NK) cell decoy. It ligates LIR-1/ILT2 (CD85j), an NK inhibitory receptor, to prevent lysis of infected target cells. However, precise role of UL18 to NK cell cytotoxicity is yet elusive. Difficulty in clarifying the function of UL18 lies in complication in detecting UL18 mainly due to low level expression of UL18 on the surface and gradual loss of its expression. METHODS: To overcome this hurdle, cDNA of cytoplasmic tail-less UL18 was constructed and expressed in swine endothelial cell (SEC). The expression level and its stability in the cell surface were monitored with FACS analysis. RESULTS: Surface expression of UL18 is up-regulated by removing cytoplasmic tail portion from UL18F (a full sequence of UL18). SECs transfected with a cDNA of UL18CY (a cytoplasmic tail-less UL18) stably expressed UL18 molecule on the surface without gradual loss of its expression during 6 week continuous cultures. In the NK cytotoxicity assay, UL18 functions either inhibiting or activating NK cell cytotoxicity according to the source of NK cells. We found that there is individual susceptibility in determining whether the engagement of NK cell and UL18 results in overall inhibiting or activating NK cell cytotoxicity. CONCLUSION: In this study, we found that cytoplasmic tail is closely related to the regulatory function for controlling surface expression of UL18. Furthermore, by constructing stable cell line in which UL18 expression is up-regulated and stable, we provided a useful tool to clarify exact functions of UL18 on various immune cells having ILT2 receptor.

Association between the level of Bcl-2 expression in EBV-LCLs and the sensitivity of NK's cytotoxicity to EBV-LCLs / 中国免疫学杂志

Objective:To investigate the relationship between the level of Bcl 2 protein in EB virus infected cells and the sensitivity of this cells to NK activity and apoptosis inducing factors.Methods:Antisense oligodexynucleotides(ODNs) were used to modulate the expression of Bcl 2 gene in Epstein Barr virus transformed B lymphoblastoid cell lines(EBV LCLs);then the change of cytotoxicity of natural killer cells and the sensitivity of apoptosis inducing elements(taking out growth factor ?dexamethasone)targeting EBV LCLs,were investigated.Results:The level of Bcl 2 expression in EBV LCLs has negative relativity with the cytotoxicity of NK cells to EBV LCLs(P