Cargando…

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 cu...

Descripción completa

Detalles Bibliográficos
Autores principales: Stefas, Ilias, Tigrett, Sylvia, Dubois, Grégor, Kaiser, Marco, Lucarz, Estelle, Gobby, Delphine, Bray, Dorothy, Ellerbrok, Heinz, Zarski, Jean Pierre, Veas, Francisco
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4621047/
https://www.ncbi.nlm.nih.gov/pubmed/26502286
http://dx.doi.org/10.1371/journal.pone.0140900
_version_ 1782397387083350016
author Stefas, Ilias
Tigrett, Sylvia
Dubois, Grégor
Kaiser, Marco
Lucarz, Estelle
Gobby, Delphine
Bray, Dorothy
Ellerbrok, Heinz
Zarski, Jean Pierre
Veas, Francisco
author_facet Stefas, Ilias
Tigrett, Sylvia
Dubois, Grégor
Kaiser, Marco
Lucarz, Estelle
Gobby, Delphine
Bray, Dorothy
Ellerbrok, Heinz
Zarski, Jean Pierre
Veas, Francisco
author_sort Stefas, Ilias
collection PubMed
description 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.
format Online
Article
Text
id pubmed-4621047
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-46210472015-10-29 Interactions between Hepatitis C Virus and the Human Apolipoprotein H Acute Phase Protein: A Tool for a Sensitive Detection of the Virus Stefas, Ilias Tigrett, Sylvia Dubois, Grégor Kaiser, Marco Lucarz, Estelle Gobby, Delphine Bray, Dorothy Ellerbrok, Heinz Zarski, Jean Pierre Veas, Francisco PLoS One Research Article 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. Public Library of Science 2015-10-26 /pmc/articles/PMC4621047/ /pubmed/26502286 http://dx.doi.org/10.1371/journal.pone.0140900 Text en © 2015 Stefas et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Stefas, Ilias
Tigrett, Sylvia
Dubois, Grégor
Kaiser, Marco
Lucarz, Estelle
Gobby, Delphine
Bray, Dorothy
Ellerbrok, Heinz
Zarski, Jean Pierre
Veas, Francisco
Interactions between Hepatitis C Virus and the Human Apolipoprotein H Acute Phase Protein: A Tool for a Sensitive Detection of the Virus
title Interactions between Hepatitis C Virus and the Human Apolipoprotein H Acute Phase Protein: A Tool for a Sensitive Detection of the Virus
title_full Interactions between Hepatitis C Virus and the Human Apolipoprotein H Acute Phase Protein: A Tool for a Sensitive Detection of the Virus
title_fullStr Interactions between Hepatitis C Virus and the Human Apolipoprotein H Acute Phase Protein: A Tool for a Sensitive Detection of the Virus
title_full_unstemmed Interactions between Hepatitis C Virus and the Human Apolipoprotein H Acute Phase Protein: A Tool for a Sensitive Detection of the Virus
title_short Interactions between Hepatitis C Virus and the Human Apolipoprotein H Acute Phase Protein: A Tool for a Sensitive Detection of the Virus
title_sort interactions between hepatitis c virus and the human apolipoprotein h acute phase protein: a tool for a sensitive detection of the virus
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4621047/
https://www.ncbi.nlm.nih.gov/pubmed/26502286
http://dx.doi.org/10.1371/journal.pone.0140900
work_keys_str_mv AT stefasilias interactionsbetweenhepatitiscvirusandthehumanapolipoproteinhacutephaseproteinatoolforasensitivedetectionofthevirus
AT tigrettsylvia interactionsbetweenhepatitiscvirusandthehumanapolipoproteinhacutephaseproteinatoolforasensitivedetectionofthevirus
AT duboisgregor interactionsbetweenhepatitiscvirusandthehumanapolipoproteinhacutephaseproteinatoolforasensitivedetectionofthevirus
AT kaisermarco interactionsbetweenhepatitiscvirusandthehumanapolipoproteinhacutephaseproteinatoolforasensitivedetectionofthevirus
AT lucarzestelle interactionsbetweenhepatitiscvirusandthehumanapolipoproteinhacutephaseproteinatoolforasensitivedetectionofthevirus
AT gobbydelphine interactionsbetweenhepatitiscvirusandthehumanapolipoproteinhacutephaseproteinatoolforasensitivedetectionofthevirus
AT braydorothy interactionsbetweenhepatitiscvirusandthehumanapolipoproteinhacutephaseproteinatoolforasensitivedetectionofthevirus
AT ellerbrokheinz interactionsbetweenhepatitiscvirusandthehumanapolipoproteinhacutephaseproteinatoolforasensitivedetectionofthevirus
AT zarskijeanpierre interactionsbetweenhepatitiscvirusandthehumanapolipoproteinhacutephaseproteinatoolforasensitivedetectionofthevirus
AT veasfrancisco interactionsbetweenhepatitiscvirusandthehumanapolipoproteinhacutephaseproteinatoolforasensitivedetectionofthevirus