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目的制备鼠抗人CC型趋化因子受体5(CCR5)特异性Ⅰ型艾滋病病毒(HIV-1)中和抗体。方法用稳定表达人CCR5的CHO-R5细胞腹腔免疫BALB/c小鼠;流式细胞术的方法分析免疫血清对GHOST-R5细胞的结合;纯化免疫血清总IgG抗体,HIV-1假病毒中和试验分析血清总IgG的中和活性。结果流式细胞术的结果显示,CHO-R5细胞免疫组小鼠血清,对GHOST-R5结合的平均荧光强度显著高于对照组(P<0.001);HIV-1假病毒中和试验的结果发现,纯化的CHO-R5细胞组小鼠免疫血清总IgG,可抑制HIV-1假病毒在体外感染GHOST-R5细胞,抑制作用具有剂量依赖性,最高抑制率达到92%。结论成功诱导出具有中和HIV-1活性的鼠抗人CCR5的多克隆抗体。
Objective To prepare mouse anti-human CCR5-specific HIV-1 neutralizing antibody. Methods BALB / c mice were immunized by intraperitoneal injection of CHO-R5 cells stably expressing human CCR5. Flow cytometry was used to analyze the binding of immune serum to GHOST-R5 cells. Total IgG antibodies against immune serum, HIV-1 pseudovirions neutralization The assay analyzes the total serum IgG neutralizing activity. Results The results of flow cytometry showed that the mean fluorescence intensity of GHOST-R5 binding in the serum of CHO-R5 cell-bearing mice was significantly higher than that of the control group (P <0.001). As a result of the HIV-1 pseudovirion neutralization test , And the total serum IgG of the purified CHO-R5 cells could inhibit the HIV-1 pseudovirus from infecting GHOST-R5 cells in vitro in a dose-dependent manner with the highest inhibition rate of 92%. Conclusions Polyclonal antibodies against murine human CCR5 that neutralize HIV-1 activity were successfully induced.