Antibody Laboratory, Protein Research Dept., Genetic Engineering and Biotechnology Research Institute, City for Scientific Research and Technology Applications, New Borg EL-Arab, Alexandria 21394, Egypt.
Virol J. 2012 Sep 16;9:201. doi: 10.1186/1743-422X-9-201.
To extend the study of the camel milk proteins which have antiviral activity against HCV, camel naïve polyclonal IgGs, α-lactalbumin were purified from camel milk and their anti-HCV effect was examined using PBMCs and Huh7.5 cell-lines. They were compared with the activity of human polyclonal IgGs and camel lactoferrin and casein.
Three types of experiments were performed on PBMCs and HuH7.5 cell. HCV was directly incubated with the purified proteins and then mixed with both cell types, or the proteins were incubated with the cells and then exposed to HCV, or the HCV pre-infected cells were treated with the proteins to inhibit intracellular replication. The proteins were added to cells or virus at different concentrations and time intervals.
Pretreated PBMCs and Huh7.5 cells with milk proteins were not protected when exposed to HCV infection. The direct interaction between HCV and camel IgGs and camel lactoferrin (cLf) led to a complete inhibition of HCV entry into cells, while casein, α-lactalbumin and human IgGs failed to inhibit HCV entry at any tested concentration. Camel IgGs showed ability to recognize HCV peptides with a significant titer (12 × 10(3)) in comparison with human IgGs which failed to do it. Camel lactoferrin was capable of inhibiting the intracellular HCV replication at concentrations of 0.25-1.25 mg/ml.
Camel milk naïve polyclonal IgGs isolated from camel milk could inhibit the HCV infectivity and demonstrated strong signal against its synthetic peptides. Lactoferrin inhibit the HCV infectivity started from 0.25 mg/ml. However, α-lactalbumin, human IgGs and casein failed to demonstrate any activity against HCV infectivity.
扩展对具有抗 HCV 活性的骆驼乳蛋白的研究,从骆驼乳中纯化骆驼天然多克隆 IgG、α-乳白蛋白,并使用 PBMC 和 Huh7.5 细胞系检查其抗 HCV 作用。将它们与人类多克隆 IgG 和骆驼乳铁蛋白和酪蛋白的活性进行比较。
在 PBMC 和 HuH7.5 细胞上进行了三种类型的实验。将 HCV 直接与纯化的蛋白质孵育,然后与两种细胞类型混合,或使蛋白质与细胞孵育,然后暴露于 HCV,或用蛋白质处理预先感染 HCV 的细胞以抑制细胞内复制。将蛋白质以不同的浓度和时间间隔添加到细胞或病毒中。
用乳蛋白预处理的 PBMC 和 Huh7.5 细胞在暴露于 HCV 感染时未得到保护。HCV 与骆驼 IgG 和骆驼乳铁蛋白(cLf)之间的直接相互作用导致 HCV 进入细胞完全被抑制,而酪蛋白、α-乳白蛋白和人 IgG 未能在任何测试浓度下抑制 HCV 进入。与未能做到这一点的人 IgG 相比,骆驼 IgG 显示出识别 HCV 肽的能力,其效价显著(12×10(3))。骆驼乳铁蛋白能够以 0.25-1.25mg/ml 的浓度抑制细胞内 HCV 复制。
从骆驼乳中分离的骆驼乳天然多克隆 IgG 可抑制 HCV 感染力,并对其合成肽显示出强烈的信号。乳铁蛋白抑制 HCV 感染力从 0.25mg/ml 开始。然而,α-乳白蛋白、人 IgG 和酪蛋白未能显示出任何针对 HCV 感染力的活性。