The human CXCR3 receptor is a G protein-coupled receptor expressed on the surface of CD 4+ cells, CD8+ cytotoxic T-cells, and natural killer T-cells. CXCR3 has been implicated in inflammation.  Although several ligands of CXCR3 have been reported, the receptor's function and signaling pathways are unclear.  This study's objective is to determine if CXCR3 mediates the function of serum amyloid A (SAA), an acute-phase plasma protein. SAA has been used as a marker for chronic inflammatory diseases, since It has been found to recruit T cells at inflammatory sites.  In order to study whether CXCR3 is a receptor of SAA,  a cell line, which  over-expresses CXCR3 on its surface, was established. Using DNA cloning technology, we subcloned hCXCR3 open reading frame into a mammalian expression vector with specific restriction enzymes. Then, CXCR3 DNA plasmid was transfected into  human epithelial kidney cells to express the hCXCR3 receptor on its surface .  The hCXCR3 receptor was detected by labeling the receptor with its specific antibody and a fluorescent conjugated secondary antibody, and then using cell flow cytomery(FACS).  The data showed that 50% - 70% of CXCR3 transfected HEK293 cells successfully expressed the receptor.  Next, we will select the receptor over-expressed cells for SAA testing.

Expression of CXCR3 receptor by HEK 293 cells transfected with recombinant plasmid DNA pCMV/hCXCR3

243rd National Meeting (2012)

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