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目的探讨调节活化正常T细胞表达与分泌趋化因子(RANTES)及单核细胞趋化蛋白-1(MCP-1)在HIV-1感染中的作用。方法以ELISA检测HIV-1感染者(治疗组和未治疗组)和健康对照组人群外周血清RANTES及MCP-1的浓度;构建hRANTES-pcDNA3.1,hMCP-pcDNA 3.1及hMCP/hRANTES-pcDNA3.1重组质粒,在CHO细胞中体外高效表达,获得重组蛋白,并研究三者对人外周血单个核细胞的趋化功能。结果RANTES在健康对照组为(164.3±21.3)pg/mL,未治疗组为(1224.1±62.0)pg/mL,治疔组为(475.(?)±36.2)pg/mL;MCP-1分别为(90.6±28.5)、(335.0±30.3)和(807.2±62.6)pg/mL。HIV-1感染后RANTES和MCP-1均升高,有效抗病毒治疗后RANTES有所下降,而MCP-1则明显上升。重组蛋白经Western印迹鉴定.能与各自单克隆抗体结合,且三者对人外周血单个核细胞都有趋化作用,MCP/RANTES融合蛋白的趋化作用更强。在重组蛋白浓度为50、200、400和800 pg/mL时,MCP/RANTES融合蛋白趋化外周血单个核细胞数分别为52×10~4/mL、102×10~4/mL、132×10~4/mL和184×10~4/mL;RANTES趋化外周血单个核细胞数分别为27×10~4/mL、51×10~4/mL、65×10~4/mL和96×10~4/mL;MCP-1趋化外周血单个核细胞数分别为18×10~4/mL、44×10~4/mL、54×10~4/mL和74×10~4/mL。结论RANTES和MCP-1可能都参与了HIV-1的感染和机体对HIV-1的免疫反应。
Objective To investigate the role of RANTES and monocyte chemotactic protein-1 (MCP-1) in the activation of HIV-1 infection. Methods Serum levels of RANTES and MCP-1 in peripheral blood of HIV-1 infected (untreated and treated) and HIV-1 infected patients were detected by ELISA. HRANTES-pcDNA3.1, hMCP-pcDNA3.1 and hMCP / hRANTES-pcDNA3 were constructed. 1 recombinant plasmid, highly expressed in CHO cells in vitro to obtain the recombinant protein, and to study the chemotactic function of the three on human peripheral blood mononuclear cells. Results RANTES was (164.3 ± 21.3) pg / mL in healthy control group, (1224.1 ± 62.0) pg / mL in untreated group and (475 ± 36.2) pg / mL in treatment group; (90.6 ± 28.5), (335.0 ± 30.3) and (807.2 ± 62.6) pg / mL, respectively. RANTES and MCP-1 increased after HIV-1 infection, RANTES decreased and MCP-1 increased significantly after effective antiviral treatment. The recombinant proteins were identified by Western blotting and could bind to their respective monoclonal antibodies, and all three had chemotactic effects on human peripheral blood mononuclear cells, and chemotactic effects of the MCP / RANTES fusion protein were stronger. The numbers of MCP-RANTES fusion protein peripheral blood mononuclear cells were 52 × 10-4 / mL, 102 × 10-4 / mL and 132 × 10 4 at the concentrations of recombinant protein of 50, 200, 400 and 800 pg / mL, respectively 10 ~ 4 / mL and 184 × 10 ~ 4 / mL respectively. The numbers of peripheral blood mononuclear cells of RANTES chemotaxis were 27 × 10-4 / mL, 51 × 10-4 / mL, 65 × 10-4 / mL and 96 × 10 ~ 4 / mL. MCP-1 chemotaxis of peripheral blood mononuclear cells were 18 × 10-4 / mL, 44 × 10-4 / mL, 54 × 10-4 / mL and 74 × 10-4 / mL. Conclusion Both RANTES and MCP-1 may be involved in the infection of HIV-1 and the immune response of HIV-1 to the body.