脐血和外周血单个核细胞培养上清影响HIV-1感染的体外实验研究

目的：体外研究脐血和外周血单个核细胞（CBMC和PBMC）培养上清对HIV-1感染的影响，为发现抗HIV-1的可溶性因子奠定基础。 方法： PHA刺激CBMC和PBMC后5 h和12 h收集上清，加入荧光标记的HIV-1ⅢB/H9和MT-2细胞培养体系中，2 h后在倒置荧光显微镜下观察其对细胞融合的影响；用Luminex 100TM分析仪检测收集上清中细胞因子含量。 结果： PHA刺激CBMC和PBMC后5 h和12 h的上清均可抑制HIV-1ⅢB/H9和MT-2细胞融合，同一时间收集的PBMC和CBMC上清对HIV-1ⅢB/H9和MT-2细胞融合的抑制作用无差异，但5 h上清的抑制作用强于12 h的上清；CBMC 5 h上清中促炎症细胞因子比PBMC 5 h上清为低，而CCR5配体MIP-1α和RANTES则比PBMC 5 h上清高，差异显著（P＜0.05）。 结论： 通过脐血和外周血单个核细胞可以制备有效抗HIV感染的可溶性因子，可能为艾滋病治疗药物提供新的来源。

Effect of supernatants from cord blood mononuclear cells and peripheral blood mononuclear cells on infection of HIV-1 in vitro

AIM: To study the effect of supernatants from cord blood mononuclear cells (CBMC) and peripheral blood mononuclear cells (PBMC) on infection of HIV-1 in vitro, and develop effectively soluble factors for human acquired immunodeficiency syndrome (AIDS) treatment. METHODS: Different supernatants from CBMC and PBMC activated by PHA for 5 hours and 12 hours were added to cell culture systems between HIV-1ⅢB/H9 cells labeled by calcein-AM and MT-2 cells, then to count the fusion under a reverted fluorescent microscope after 2 hours, respectively, and different soluble factors in supernatants were detected by Luminex 100TM. RESULTS: These supernatants from CBMC and PBMC activated by PHA for 5 hours and 12 hours inhibited the formation of fusion, and there is no difference between supernatants collected in CBMC and PBMC at same time point. However, the supernatants collected in 5 hours were more effective to inhibit the formation of fusion than those in 12 hours (P<0.05). There was a lower concentration for pro-inflammatory cytokines, such as IFN-γ, MCP-1 and TNF-α, in supernatants from CBMC than those from PBMC, but the concentration of MIP-1a and RANTES in supernatants from CBMC was higher than those from PBMC. CONCLUSION: The soluble factors for anti-HIV can be prepared from CBMC and PBMC, which is a new source of anti-HIV therapeutic preparation.