Immune Netw.  2008 Dec;8(4):130-136. 10.4110/in.2008.8.4.130.

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

• Kim JS
• Kim B
• Yoon IH
• Kim SJ
• Park CG

• KMID: 2054259
• DOI: http://doi.org/10.4110/in.2008.8.4.130

Abstract

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.

Keyword

UL18; cytoplasmic tail-less UL18; NK cytotoxicity

MeSH Terms

Cell Line
Cytomegalovirus
Cytoplasm
DNA, Complementary
Endothelial Cells
Humans
Killer Cells, Natural
Swine
DNA, Complementary