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目的 研究人 β趋化因子同源物MC1 4 8在腺病毒重组体的表达以及对趋化因子MCP 1的拮抗作用。方法 从传染性软疣病毒 (MCV)基因组中克隆MC1 4 8DNA片段 ,通过同源重组构建Ad MC1 4 8腺病毒重组体。重组体Ad MC1 4 8转染 2 93细胞 ,从上清中提纯重组蛋白MC1 4 8P。通过趋化抑制实验 ,研究MC1 4 8P是否具有拮抗MCP 1趋化单核细胞的作用。结果 成功克隆MC1 4 8基因 ,经测序证明与GenBank中报道的完全相同 ;构建了复制缺陷型腺病毒重组体Ad MC1 4 8;重组蛋白MC1 4 8P终浓度分别为 :0 μg/L、1 0 μg/L、2 0 μg/L、5 0 μg/L时 ,MCP 1对单核细胞的趋化指数分别为 :1 0 0 %、4 6 .2 5 %、32 .5 7%、6 .6 7%。结论 MC1 4 8P蛋白具有拮抗和抑制趋化因子MCP 1对单核细胞的趋化作用 ,属于一种趋化因子拮抗剂 ,并且这种拮抗作用具有剂量依赖性。
Objective To study the expression of human β-chemokine homologue MC1 4 8 in adenovirus recombinant and the antagonism of chemokine MCP 1. Methods MC1 48 DNA fragment was cloned from the genome of molluscum contagiosum (MCV) and recombinant adenovirus Ad MC148 was constructed by homologous recombination. Recombinant Ad MC1 4 8 was transfected into 293 cells and the recombinant protein MC1 4 8P was purified from the supernatant. Through chemotaxis inhibition experiments, we investigated whether MC1 4 8P antagonized MCP 1 chemotaxis to monocytes. Results The MC1 48 gene was successfully cloned and sequenced to be identical with that reported in GenBank. The recombinant adenovirus Ad MC1 48 was constructed and the final concentration of recombinant MC1 48P was 0 μg / L, 10 The chemotactic index of MCP 1 to monocytes was 100%, 46.5% and 32.57%, respectively, when the concentration was 50 μg / L, 20 μg / L, 50 μg / L. 6 7%. Conclusion MC1 4 8P protein antagonizes and inhibits the chemotactic effect of chemokine MCP 1 on monocytes. It belongs to a chemokine antagonist, and the antagonism is dose-dependent.