Improved reactivity of hepatitis C virus core protein epitopes in a conformational antigen-presenting system
Improved reactivity of hepatitis C virus core protein epitopes in a conformational antigen-presenting system
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DOI：
10.1128/cdli.4.2.117-121.1997
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发表时间：
1997-03-01
期刊：
影响因子：
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通讯作者：
Tisminetzky, SG
中科院分区：
文献类型：
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作者：
Buratti, E;DiMichele, M;Tisminetzky, SG
最近的研究已经鉴定了核衣壳蛋白N-末端部分的几个表位，这些表位主要被丙型肝炎病毒（HCV）感染患者的血清识别。被这些抗体识别的序列的表征和它们的反应性的评价主要用合成肽进行。然而，当免疫应答针对构象表位时，合成肽作为抗原是众所周知的不可靠的。为了改善HCV感染患者中抗体应答的检测，我们评估了HCV核心蛋白三个免疫优势区域的反应性（残基1至20、21至40和32至46）以构象特异性方式展示在鸡群屋病毒（FHV）衣壳蛋白的表面上。用这些蛋白质分析94份抗HCV酶联免疫吸附试验阳性的血清样品，然后与相应的合成肽得到的结果进行了比较，与肽和FHV蛋白最反应的序列是从残基1至20的区域，证实了这些残基的展示的低构象要求。另一方面，已经报道的残基32至46的构象性质与当由FHV重组蛋白展示时观察到的其高反应性和由其相应的合成肽展示的低反应性一致。最后，对于展示残基21至40的区域的嵌合蛋白观察到的高反应性，与用合成肽获得的结果相反，还表明该序列含有一个或多个构象表位，其结构不能用合成肽正确模拟。
Recent studies have identified several epitopes in the N-terminal portion of the nucleocapsid protein which are predominantly recognized by sera of patients infected with hepatitis C virus (HCV). The characterization of the sequences recognized by these antibodies and the evaluation of their reactivities have been performed mainly with synthetic peptides, However, synthetic peptides are notoriously unreliable as antigens when the immune response is directed against conformational epitopes, In order to improve the detection of antibody responses in HCV-infected patients, we have evaluated the reactivities of three immunodominant regions of the HCV core protein (residues 1 to 20, 21 to 40, and 32 to 46) displayed in a conformation-specific manner on the surface of the Flock House virus (FHV) capsid protein. The results obtained with these proteins in the analysis of 94 serum samples positive by anti-HCV enzyme-linked immunosorbent assay were then compared with those obtained with the corresponding synthetic peptides, The sequence most reactive both with the peptide and with the FHV protein was the region from residues 1 to 20, confirming the low conformational requirements for the display of these residues. On the other hand, the already reported conformational nature of residues 32 to 46 is in keeping with its observed high reactivity when displayed by the FHV recombinant protein and with the low reactivity displayed by its corresponding synthetic peptide, Finally, the high reactivity observed for the chimeric protein displaying the region from residues 21 to 40, as opposed to the results obtained with the synthetic peptide, also suggests that this sequence contains one or more conformational epitopes whose structures cannot be mimicked correctly with synthetic peptides.