Interactions between Hepatitis C Virus and the Human Apolipoprotein H Acute Phase Protein: A Tool for a Sensitive Detection of the Virus.
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.
ApoH-Technologies, Faculté de Pharmacie, Université de Montpellier, Montpellier, France.
Ilias
I
Stefas
ApoH-Technologies, Faculté de Pharmacie, Université de Montpellier, Montpellier, France; Institut de Recherche pour le Développement, UMR-Ministère de la Défense 3, Laboratoire d'Immuno-Physiopathologie Moléculaire Comparée, Faculté de Pharmacie, Montpellier, France.
Sylvia
S
Tigrett
Institut de Recherche pour le Développement, UMR-Ministère de la Défense 3, Laboratoire d'Immuno-Physiopathologie Moléculaire Comparée, Faculté de Pharmacie, Montpellier, France.
Grégor
G
Dubois
GenExpress GmbH, Berlin, Germany.
Marco
M
Kaiser
ApoH-Technologies, Faculté de Pharmacie, Université de Montpellier, Montpellier, France.
Estelle
E
Lucarz
ApoH-Technologies, Faculté de Pharmacie, Université de Montpellier, Montpellier, France.
Delphine
D
Gobby
Immunoclin Corporation, Washington, DC, United States of America.
Dorothy
D
Bray
Robert Koch-Institute, Centre for Biological Threats and Special Pathogens, Highly Pathogenic Viruses, Berlin, Germany.
Heinz
H
Ellerbrok
Clinique d'Hépato-gastroentérologie, Centre Hospitalier Universitaire de Grenoble, IAB, INSERM U823, Grenoble, France.
Jean Pierre
JP
Zarski
Institut de Recherche pour le Développement, UMR-Ministère de la Défense 3, Laboratoire d'Immuno-Physiopathologie Moléculaire Comparée, Faculté de Pharmacie, Montpellier, France.
Francisco
F
Veas
D000209:Acute-Phase Proteins / Q000276:immunology / Q000378:metabolism*; D002499:Centrifugation, Density Gradient / Q000379:methods; D005188:False Negative Reactions; D016174:Hepacivirus / Q000276:immunology / Q000378:metabolism*; D006526:Hepatitis C / Q000175:diagnosis*; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods; D008854:Microscopy, Electron; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D053482:beta 2-Glycoprotein I / Q000276:immunology / Q000378:metabolism*
D000209:Acute-Phase Proteins; D053482:beta 2-Glycoprotein I