Expression Of CXCR4 In Shrews And Human Lymphocytes And Its Chemotactic Function

Objective:To study the CXCR4gene expression and cell migration of lymphocytes induced by rapacymin, we isolated the cells from tree shrew and human.Methods:Firstly, we established and developed three quick protocols to isolate and analyze the cells from peripheral blood, bone and spleen tissues. Secondly, we isolated and treated the tree shrew’s cells with rapacymin. The survival of cell was tested by the MTS. Migration assays were performed in AP48Well Micro Chemotaxis Chamber with Filters for AP48. Real-time PCR was performed to find the expression of CXCR4gene. Thirdly, we isolated and treated the human’s cells with rapacymin. Then migration assays was used to find the chemotaxis of the cells and the real-timePCR was performed to find the expression of CXCR4gene, compared to the tree shrew’s.Reasults:Results from flow cytometry analysis further confirmed the successful isolation of tree shrew’s lymphocytes. The MTS results showed that rapacymin had little effect on the survival of tree shrew’s lymphocytes. Migration assays showed rapacymin can inhibite the migration of the lymphocytes and reduce the expression of CXCR4gene.Conclusion:1. Rapacymin’s concentration has little effect on the survival of lymphocyte.2. The10ng/ml rapacymin is effective in inhibition of both tree shrew and human’s lymphocyte’s migration by CXCL12.3. Rapacymin might affect CXCL12/CXCR4signaling pathways through down-regulating CXCR4gene expression and inhibit the lymphocyte’s migration.