Active Chemokine (C-X-C Motif) Ligand 14 (meiceguo ) is a member of the CXC chemokine superfamily (1-5). CXCL14 is expressed by a variety of immune and non-immune cells,such as monocytes and B cells stimulated by LPS. It has been demonstrated that CXCL14 is a highly selective chemoattractant for monocytes that have been treated with prostaglandin E2 or forskolin, agents that activate adenylate cyclase.
It also attracts certain cells of the immune system, including dendritic cells and antigen-engaged B cells, CCR7 central-memory T-Cells. Thus, chemotaxis assay used 24-well microchemotaxis system was undertaken to detect the chemotactic effect of recombinant human CXCL14 on the THP-1 cell line. Briefly, THP-1 cells were seeded into the upper chambers (200μL cell suspension,106 cells/mL in RPMI 1640 with FBS free) and CXCL14 (0.01ng/mL, 0.1ng/mL, 1ng/mL, 10ng/mL,100ng/mL and 1000 ng/mL diluted separately in serum free RPMI 1640 ) was added in lower chamber with a polycarbonate filter (8 μm pore size) used to separate the two compartments. After incubation at 37℃ with 5% CO2 for 2h, the filter was removed, then cells in low chamber were observed by inverted microscope at low magnification (×10) and the number of migrated cells were counted randomly (five fields for each filter). Result shows MIP-3α is able to induce migration of THP-1 cells. The migrated THP-1 cells in low chamber at low magnification (×10) were shown in Figure 1. Five fields of each chamber were randomly chosen, and the migrated cells were counted at magnification (×10). Statistical results were shown in Figure 2. The optimum chemotaxis of recombinant human CXCL14 occurs at 1-1000 ng/mL.