Abstract
The Hepatitis C virus (HCV) infection exhibits a high global prevalence frequently associated with hepatocellular carcinoma, taking years to develop. Despite the standardization of highly sensitive HCV quantitative RT-PCR (qRT-PCR) detection methods, false-negative diagnoses may be generated with current methods, mainly due to the presence of PCR inhibitors and/or low viral loads in the patient’s sample. These false-negative diagnoses impact both public health systems, in developing countries, and an in lesser extent, in developed countries, including both the risk of virus transmission during organ transplantation and/or blood transfusion and the quality of the antiviral treatment monitoring. To adopt an appropriate therapeutic strategy to improve the patient’s prognosis, it is urgent to increase the HCV detection sensitivity. Based upon previous studies on HBV, we worked on the capacity of the scavenger acute phase protein, Apolipoprotein H (ApoH) to interact with HCV. Using different approaches, including immunoassays, antibody-inhibition, oxidation, ultracentrifugation, electron microscopy and RT-PCR analyses, we demonstrated specific interactions between HCV particles and ApoH. Moreover, when using a two-step HCV detection process, including capture of HCV by ApoH-coated nanomagnetic beads and a home-made real-time HCV-RT-PCR, we confirmed the presence of HCV for all samples from a clinical collection of HCV-seropositive patients exhibiting an RT-PCR COBAS® TaqMan® HCV Test, v2.0 (COBAS)-positive result. In contrast, for HCV-seropositive patients with either low HCV-load as determined with COBAS or exhibiting HCV-negative COBAS results, the addition of the two-step ApoH-HCV-capture and HCV-detection process was able to increase the sensitivity of HCV detection or more interestingly, detect in a genotype sequence-independent manner, a high-proportion (44%) of HCV/RNA-positive among the COBAS HCV-negative patients. Thus, the immune interaction between ApoH and HCV could be used as a sample preparation tool to enrich and/or cleanse HCV patient’s samples to enhance the detection sensitivity of HCV and therefore significantly reduce the numbers of false-negative HCV diagnosis results.
Highlights
Until the recent introduction of hepatitis C virus (HCV) screening tests, this viral infection has represented the major cause of blood transfusion-associated hepatitis [1]
We have evidenced that the scavenger acute phase protein Apolipoprotein H (ApoH) was able to capture RNA-containing HCV particles from infected patients’ sera independently of the tested genotypes
This binding was specific as an anti-ApoH MAb was able to almost totally abrogate this interaction, whereas no effect was observed in the presence of α1 acid-glycoprotein, another acute phase plasma protein
Summary
Until the recent introduction of hepatitis C virus (HCV) screening tests, this viral infection has represented the major cause of blood transfusion-associated hepatitis [1]. More than 70% of the HCV-infected individuals develop a chronic infection considered as a major cause of liver cirrhosis and hepatocellular carcinoma [3]. Other lympho-proliferative disorders may be associated with HCV infection, including mixed cryoglobulinemia (MC) and Non-Hodgkin’s lymphoma (NHL) [4]. The HCV pathogenesis is not well understood, viral infection progresses slowly and often ends in chronic diseases. HCV mainly targets the liver cells [5], but this virus may replicate in extra-hepatic cells such as T, B and monocyte cell subsets from chronically infected individuals [6]
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