Lu Yinping, Xu Yang, Yang Dongliang, Kemper Thekla, Roggendorf Michael, Lu Mengji
Institut für Virologie, Universitätsklinikum Essen, Hufeland str. 55, 45122 Essen, Germany.
Cytokine. 2008 Feb;41(2):127-35. doi: 10.1016/j.cyto.2007.11.002. Epub 2007 Dec 21.
Interferon (IFN)-alpha and -beta are important antiviral mediators. IFN-alpha is widely used for the treatment of chronic hepatitis B. In our previous studies, a subtype of woodchuck IFN-alpha (wIFN-alpha) was characterized and has been shown to be active in suppressing the replication of woodchuck hepatitis virus (WHV) in vitro and vivo. Here, we refined the analysis of the IFN-alpha/beta system of the woodchuck and studied the expression of wIFN-alpha/beta in peripheral blood lymphocytes (PBLs) from naïve and WHV-infected woodchucks. A number of wIFN-alpha genes were sequenced and could be classified into 10 subtypes and 3 pseudotypes. The biological activity of different subtypes of wIFN-alpha was demonstrated by their ability to protect woodchuck cells against encephalomyocarditis virus infection and to induce MxA expression in woodchuck cells. Additionally, a partial sequence of wIFN-beta was characterized. A subtyping method for wIFN-alpha based on restriction length polymorphism analysis was developed. Further, the expression of wIFN by woodchuck PBLs after stimulation with polyI/C was investigated. The maximal production of wIFN by woodchuck PBLs occurred within the first 48 h after addition poly I/C. The wIFN-alpha subtypes 1, 4, and 5 were found to be produced by poly I/C-stimulated woodchuck PBLs, indicating a selective expression of wIFN-alpha subtypes. PBLs from chronically WHV-infected woodchucks showed a reduced ability to produce wIFN when stimulated with poly I/C. The results suggest that woodchucks with chronic WHV infection have impaired immunological responses to poly I/C.
干扰素（IFN）-α和-β是重要的抗病毒介质。IFN-α被广泛用于治疗慢性乙型肝炎。在我们之前的研究中，土拨鼠IFN-α（wIFN-α）的一个亚型已被鉴定，并且已证明其在体外和体内抑制土拨鼠肝炎病毒（WHV）复制方面具有活性。在此，我们完善了对土拨鼠IFN-α/β系统的分析，并研究了wIFN-α/β在未感染和感染WHV的土拨鼠外周血淋巴细胞（PBL）中的表达。对多个wIFN-α基因进行了测序，可分为10个亚型和3个假型。不同亚型的wIFN-α的生物活性通过其保护土拨鼠细胞免受脑心肌炎病毒感染以及诱导土拨鼠细胞中Mx A表达的能力得以证明。此外，还鉴定了wIFN-β的部分序列。开发了一种基于限制性长度多态性分析的wIFN-α亚型分型方法。此外，还研究了用聚肌胞苷酸（polyI/C）刺激后土拨鼠PBL中wIFN的表达。土拨鼠PBL产生wIFN的最大值出现在添加polyI/C后的前48小时内。发现polyI/C刺激后土拨鼠PBL产生wIFN-α亚型1、4和5，表明wIFN-α亚型存在选择性表达。来自慢性感染WHV的土拨鼠的PBL在用polyI/C刺激时产生wIFN的能力降低。结果表明，慢性感染WHV的土拨鼠对polyI/C的免疫反应受损。
