Department of Medical Microbiology, Medical College, Qingdao University, Qingdao 266071, China.
Virol Sin. 2010 Dec;25(6):432-9. doi: 10.1007/s12250-010-3131-3. Epub 2010 Dec 21.
In this study, a standard strain of HSV-1 (strain SM(44)) was used to investigate the antiviral activity of the recombinant Cyanovirin-N (CV-N) against Herpes simplex virus type 1 (HSV-1) in vitro and in vivo. Cytopathic effect (CPE) and MTT assays were used to evaluate the effect of CV-N on HSV-1 in Vero cells. The number of copies of HSV-DNA was detected by real-time fluorescence quantitative PCR (FQ-PCR). The results showed that CV-N had a low cytotoxicity on Vero cells with a CC(50) of 359.03 ± 0.56 μg/mL, and that it could not directly inactivate HSV-1 infectivity. CV-N not only reduced the CPE of HSV-1 when added before or after viral infection, with a 50% inhibitory concentration (IC(50)) with 2.26 and 30.16 μg/mL respectively, but it also decreased the copies of HSV-1 DNA in infected host cells. The encephalitis model for HSV-1 infection was conducted in Kunming mice, and treated with three dosages of CV-N (0.5, 5 & 10 mg/kg) which was administered intraperitoneally at 2h, 3d, 5d, 7d post infection. The duration for the appearance of symptoms of encephalitis and the survival days were recorded and brain tissue samples were obtained for pathological examination (HE staining). Compared with the untreated control group, in the 5mg/kg CV-N and 10mg/kg CV-N treated groups, the mice suffered light symptoms and the number of survival days were more than 9 d and 14 d respectively. HE staining also showed that in 5mg/kg CV-N and 10mg/kg CV-N treated groups, the brain cells did not show visible changes, except for a slight inflammation. Our results demonstrated that CV-N has pronounced antiviral activity against HSV-1 both in vitro and in vivo, and it would be a promising new candidate for anti-HSV therapeutics.
在这项研究中,使用标准的 HSV-1 株(SM(44)株)来研究重组 Cyanovirin-N(CV-N)对单纯疱疹病毒 1 型(HSV-1)的抗病毒活性。细胞病变效应(CPE)和 MTT 试验用于评估 CV-N 在 Vero 细胞中对 HSV-1 的作用。通过实时荧光定量 PCR(FQ-PCR)检测 HSV-DNA 的拷贝数。结果表明,CV-N 对 Vero 细胞的细胞毒性较低,CC(50)为 359.03 ± 0.56 μg/mL,并且不能直接灭活 HSV-1 感染性。CV-N 不仅在病毒感染前后加入时减少 HSV-1 的 CPE,IC(50)分别为 2.26 和 30.16 μg/mL,而且还降低了感染宿主细胞中 HSV-1 DNA 的拷贝数。在昆明小鼠中进行 HSV-1 感染性脑炎模型,并使用三种剂量的 CV-N(0.5、5 和 10 mg/kg)进行腹腔内给药,在感染后 2h、3d、5d、7d 给药。记录脑炎症状出现的持续时间和存活天数,并获取脑组织样本进行病理检查(HE 染色)。与未处理的对照组相比,在 5mg/kg CV-N 和 10mg/kg CV-N 治疗组中,小鼠表现出较轻的症状,存活天数超过 9d 和 14d。HE 染色还显示,在 5mg/kg CV-N 和 10mg/kg CV-N 治疗组中,脑细胞除了轻微炎症外,没有明显变化。我们的结果表明,CV-N 对 HSV-1 具有显著的体外和体内抗病毒活性,是一种有前途的抗 HSV 治疗新候选药物。