Background: HCV Alternate Reading Frame Protein (ARFP) is a frameshift product of HCV-core encoding. Here, we characterized specific anti-ARFP antibodies in Liver Transplant Candidate (LTC) and chronic HCV-infected patients.

Methods: The ARFP gene was cloned and the recombinant protein was purified using Nickel chromatography and confirmed by western blotting. ELISA was developed using recombinant core-1a, core- 1b, ARFP-1a protein, and 99-residue synthetic ARFP 1b peptide. By several Bioinformatics tools, general properties, immunogenic epitopes, and structures of these proteins were obtained.

Results: The seroprevalence of anti-core and anti-ARFP antibodies was 100% in LTC patients, but only 75.2% and 94.3% of chronic patients had evidence of anti-ARFP and anti-core antibodies, respectively. In-silico results demonstrated physicochemical features, antigen properties and potential interactors that could describe progression toward advanced liver disease.

Conclusion: As the first report, the prevalence of anti-ARFP antibodies in LTC patients is of the order of 100% and titer of anti-ARFP antibody was significantly higher in LTC patients compared to chronic individuals, suggesting the possible role of ARFP in the progression toward advanced liver disease. In addition, docking analysis determined several interactor proteins such as prefoldin 2, cathepsin B, vitronectin, and angiotensinogen that have an important role in progression to chronic infection and liver disease development.

背景：HCV替代阅读框蛋白（ARFP）是HCV核心编码的移码产物。在这里，我们表征了肝移植候选（LTC）和慢性HCV感染患者的特异性抗ARFP抗体。

方法：克隆ARFP基因，用镍色谱法纯化重组蛋白，并经western blotting证实。使用重组核心-1a，核心-1b，ARFP-1a蛋白和99个残基的合成ARFP 1b肽开发了ELISA。通过几种生物信息学工具，可以获得这些蛋白质的一般性质，免疫原性表位和结构。

结果：在LTC患者中，抗核心抗体和抗ARFP抗体的血清阳性率为100％，但分别只有75.2％和94.3％的慢性患者具有抗ARFP和抗核心抗体的证据。硅内结果表明，其理化特性，抗原特性和潜在的相互作用因子可描述晚期肝病的进展。

结论：作为第一份报告，与慢性个体相比，LTC患者中抗ARFP抗体的患病率约为100％，并且抗ARFP抗体的滴度在LTC患者中显着更高，这表明ARFP可能在慢性肾病中发挥作用。向晚期肝病发展。此外，对接分析确定了几种相互作用蛋白，例如前折叠素2，组织蛋白酶B，玻连蛋白和血管紧张素原，它们在向慢性感染和肝病发展的过程中具有重要作用。