Effect of TAP-SSL5 fusion protein on binding of activated platelets to human lymphocytes |
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Authors: | PENG Song BEI Jun-jie HU Hou-yuan CHEN Qiang |
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Affiliation: | Department of Cardiology, Southwest Hospital, Third Military Medical University, Chongqing 400038, China |
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Abstract: | AIM: To study the effect of tick anticoagulant peptide-staphylococcal superantigen like protein 5 (TAP-SSL5), an anti-inflammatory and anticoagulant fusion protein, on the binding of activated platelets to human lymphocytes.METHODS: Human periphery lymphocytes were isolated by magnetic activated cell sorting (MACS). The toxicity of TAP-SSL5 on the viability of Jurkat cell was assessed by CCK-8 assay. Flow cytometry was applied to detect the expression of CD162 (PSGL-1) on the Jurkat cells (human peripheral blood leukemia T lymphocyte cell line) and the inhibitory effect of TAP-SSL5 on the binding of mouse anti-human CD162 monoclonal antibody (KPL-1) to Jurkat cells. Platelets were activated by ADP at concentration of 20 μmol/L, the binding rates of activated platelets to Jurkat cells or human lymphocytes were assayed by flow cytometry. RESULTS: The concentration of TAP-SSL5 below 30 mg/L didn't affect the viability of Jurkat cells. TAP-SSL5 at 10 mg/L competitively inhibited KPL-1 binding to Jurkat cells. The binding rates of activated platelets to Jurkat cells or lymphocytes were (11.86±4.49)% and (8.32±1.00)%, respectively, which decreased to (6.73±2.71)% and (5.51±0.70)% after the Jurkat cells and lymphocytes were pre-incubated with 10 mg/L TAP-SSL5 (P <0.05).CONCLUSION: TAP-SSL5 binds to PSGL-1 expressed on lymphocyte surface and directly inhibits the binding of activated platelets to human lymphocytes, which may be one of the anti-inflammatory mechanisms of TAP-SSL5. |
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Keywords: | Tick anticoagulant peptide Staphylococcal superantigen like protein 5 Fusion protein Platelets Lymphocytes P-selectin glycoprotein ligand 1 |
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