淋巴细胞HIV-1辅受体表型剔除阻断病毒感染的研究

目的　观察HIV 1辅受体CCR5和CXCR4表型剔除对HIV 1DP1株感染的阻断作用。方法　用含有pLNCX R K S K的重组逆转录病毒液感染原代人PBLs(外周血淋巴细胞 ) ,抗体 免疫磁珠法分离筛选转化成功PBLs,流式细胞仪检测筛选效率 ;HIV 1DP1株攻击转化PBLs ,进行合胞体形成和p2 4抗原分泌检测。结果　抗 NGFR 免疫磁珠法获得了转化成功的PBLs ,流式细胞仪检测发现pLNCX R K S K转染组 77.4%的PBLsNGFR(神经生长因子受体 )标记物为阳性 ;HIV 1DP1株攻击后 ,未转染组和pLNCX转染组可以见到明显的合胞体形成 ,而在pLNCX R K S K转化PBLs没有见到合胞体形成 ;pLNCX R K S K转染组在感染后第 4、7和 10天皆可发现显著的p2 4抗原分泌抑制 ,抑制率分别为 15%、43 %、19%。结论　细胞内趋化因子通过与CCR5和CXCR4细胞内结合 ,使HIV 1两类主要辅受体难以在PBLs表面表达 ,从而可以达到阻断HIV 1病毒感染的目的

Phenotypic knockout of HIV-1 chemokine coreceptor CXCR4 and CCR5 by intrakines to block HIV-1 infection

Objective To investigate the impact of phenotypi c knockout of HIV-1 chemokine coreceptor CXCR4 and CCR5 by intrakines on HIV-1 infection. Methods Human PBLs were transduced with the r ecombinant vector pLNCX-R-K-S-K, followed by anti-NGFR/anti-IgG-magnetic bead selection and FCM detection. The transduced PBLs were infected with DP1 HIV -1 virus thereafter Envelop-mediated syncytium formation and p24 detection wer e carried out to identify anti-HIV-1 activity of the co-inactivation of CCR5 and CXCR4. Results pLNCX-R-K-S-K-transduced PBLs wer e isolated with an anti-NGFR/anti-IgG-magnetic bead method. After isolation, about 70% of the PBLs were positive for the NGFR marker. When transduced PBLs we re infected with DP1 HIV-1 virus, Envelop-mediated syncytium formation was alm ost completely inhibited by pLNCX-R-K-S-K transfection. Also, little p24 ant igen was found in the cultures of pLNCX-R-K- S-K-transduced PBLs. pLNCX-R -K-S-K transduction inhibited the secretion of DP1 p24 antigen by 15%, 43% an d 19% on day 4, 7 and 10 respectively. Conclusion The lym phocytes with the phenotypic knockout of CCR5 and CXCR4 could be protected from DP1 HIV-1 virus infection.