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  1. Article ; Online: Effect of High Hydrostatic Pressure Processing and Holder Pasteurization of Human Milk on Inactivation of Human Coronavirus 229E and Hepatitis E Virus.

    Bouquet, Peggy / Alexandre, Virginie / De Lamballerie, Marie / Ley, Delphine / Lesage, Jean / Goffard, Anne / Cocquerel, Laurence

    Viruses

    2023  Volume 15, Issue 7

    Abstract: In preterm infants, sterilized donor milk (DM) is frequently used for feeding when breast milk is lacking. Most human milk banks use the Holder pasteurization method (HoP) to ensure the microbiological safety of DM. However, this method degrades many ... ...

    Abstract In preterm infants, sterilized donor milk (DM) is frequently used for feeding when breast milk is lacking. Most human milk banks use the Holder pasteurization method (HoP) to ensure the microbiological safety of DM. However, this method degrades many bioactive factors and hormones. Recently, high hydrostatic pressure (HHP) processing, which preserves bioactive factors in human milk, has been proposed as an alternative method to ensure the safety of DM. Although HHP treatment has been shown to be effective for viral inactivation, the effect of HHP on viruses that may be present in the complex nutritional matrix of human milk has not yet been defined. In the present study, we compared the efficacy of two HHP protocols (4 cycles at 350 MPa at 38 °C designated as 4xHP350 treatment, and 1 cycle at 600 MPa at 20 °C designated as 1xHP600 treatment) with the HoP method on artificially virus-infected DM. For this purpose, we used human coronavirus 229E (HCoV-229E) and hepatitis E virus (HEV) as surrogate models for enveloped and non-enveloped viruses. Our results showed that HCoV-229E is inactivated by HHP and HoP treatment. In particular, the 4xHP350 protocol is highly effective in inactivating HCoV-229E. However, our results demonstrated a matrix effect of human milk on HCoV-229E inactivation. Furthermore, we demonstrated that HEV is stable to moderate pressure HHP treatment, but the milk matrix does not protect it from inactivation by the high-pressure HHP treatment of 600 MPa. Importantly, the complex nutritional matrix of human milk protects HEV from inactivation by HoP treatment. In conclusion, we demonstrated that HHP and HoP treatments do not lead to complete inactivation of both surrogate virus models, indicating that these treatments cannot guarantee total viral safety of donor milk.
    MeSH term(s) Infant ; Female ; Humans ; Infant, Newborn ; Milk, Human ; Pasteurization/methods ; Coronavirus 229E, Human ; Hepatitis E virus ; Hydrostatic Pressure ; Infant, Premature
    Language English
    Publishing date 2023-07-18
    Publishing country Switzerland
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2516098-9
    ISSN 1999-4915 ; 1999-4915
    ISSN (online) 1999-4915
    ISSN 1999-4915
    DOI 10.3390/v15071571
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Le virus de l’hépatite E - Un virus méconnu qui se dévoile.

    Ankavay, Maliki / Dubuisson, Jean / Cocquerel, Laurence

    Medecine sciences : M/S

    2019  Volume 34, Issue 12, Page(s) 1071–1078

    Abstract: The first cause of acute hepatitis in the world is due to the hepatitis E virus (HEV). This infection has long been considered as a problem only affecting developing countries. However, since the identification of zoonotic forms at the end of the last ... ...

    Title translation The hepatitis E virus, an unknown virus that reveals itself.
    Abstract The first cause of acute hepatitis in the world is due to the hepatitis E virus (HEV). This infection has long been considered as a problem only affecting developing countries. However, since the identification of zoonotic forms at the end of the last century, it has become clear that this infection also affects industrialized countries. The recent renewed interest in HEV has revealed some particularities in this virus. Indeed, although considered as a non-enveloped virus, the HEV viral particle is surrounded by a lipid membrane in the bloodstream. In addition, HEV secretes abundantly into the bloodstream non-infectious forms of its capsid protein that could serve as an immunological bait. This review summarizes recent advances on this virus for which the number of diagnosed cases increases every year.
    MeSH term(s) Animals ; Communicable Diseases, Emerging/epidemiology ; Communicable Diseases, Emerging/virology ; Developed Countries/statistics & numerical data ; Hepatitis E/epidemiology ; Hepatitis E/virology ; Hepatitis E virus/pathogenicity ; Hepatitis E virus/physiology ; History, 20th Century ; History, 21st Century ; Humans ; Zoonoses/epidemiology ; Zoonoses/virology
    Language French
    Publishing date 2019-01-09
    Publishing country France
    Document type Historical Article ; Journal Article ; Review
    ZDB-ID 632733-3
    ISSN 1958-5381 ; 0767-0974
    ISSN (online) 1958-5381
    ISSN 0767-0974
    DOI 10.1051/medsci/2018299
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: SRFBP1, an Additional Player in HCV Entry.

    Fénéant, Lucie / Cocquerel, Laurence

    Trends in microbiology

    2015  Volume 23, Issue 10, Page(s) 590–593

    Abstract: The tetraspanin CD81 dynamics and interactions with other proteins are essential for hepatitis C virus (HCV) entry. Recently, Gerold and collaborators used a proteomic approach and found the serum response factor binding protein 1 (SRFBP1) to be involved ...

    Abstract The tetraspanin CD81 dynamics and interactions with other proteins are essential for hepatitis C virus (HCV) entry. Recently, Gerold and collaborators used a proteomic approach and found the serum response factor binding protein 1 (SRFBP1) to be involved in a post-fusion entry process by interacting with CD81 upon HCV infection.
    MeSH term(s) Hepacivirus/physiology ; Humans ; Proteomics ; Transcription Factors/metabolism ; Virus Internalization
    Chemical Substances Transcription Factors
    Language English
    Publishing date 2015-10
    Publishing country England
    Document type Comment ; Journal Article
    ZDB-ID 1158963-2
    ISSN 1878-4380 ; 0966-842X
    ISSN (online) 1878-4380
    ISSN 0966-842X
    DOI 10.1016/j.tim.2015.08.003
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: The phosphatidylserine receptor TIM1 promotes infection of enveloped hepatitis E virus.

    Corneillie, Laura / Lemmens, Irma / Montpellier, Claire / Ferrié, Martin / Weening, Karin / Van Houtte, Freya / Hanoulle, Xavier / Cocquerel, Laurence / Amara, Ali / Tavernier, Jan / Meuleman, Philip

    Cellular and molecular life sciences : CMLS

    2023  Volume 80, Issue 11, Page(s) 326

    Abstract: The hepatitis E virus (HEV) is an underestimated RNA virus of which the viral life cycle and pathogenicity remain partially understood and for which specific antivirals are lacking. The virus exists in two forms: nonenveloped HEV that is shed in feces ... ...

    Abstract The hepatitis E virus (HEV) is an underestimated RNA virus of which the viral life cycle and pathogenicity remain partially understood and for which specific antivirals are lacking. The virus exists in two forms: nonenveloped HEV that is shed in feces and transmits between hosts; and membrane-associated, quasi-enveloped HEV that circulates in the blood. It is suggested that both forms employ different mechanisms for cellular entry and internalization but little is known about the exact mechanisms. Interestingly, the membrane of enveloped HEV is enriched with phosphatidylserine, a natural ligand for the T-cell immunoglobulin and mucin domain-containing protein 1 (TIM1) during apoptosis and involved in 'apoptotic mimicry', a process by which viruses hijack the apoptosis pathway to promote infection. We here investigated the role of TIM1 in the entry process of HEV. We determined that HEV infection with particles derived from culture supernatant, which are cloaked by host-derived membranes (eHEV), was significantly impaired after knockout of TIM1, whereas infection with intracellular HEV particles (iHEV) was unaffected. eHEV infection was restored upon TIM1 expression; and enhanced after ectopic TIM1 expression. The significance of TIM1 during entry was further confirmed by viral binding assay, and point mutations of the PS-binding pocket diminished eHEV infection. In addition, Annexin V, a PS-binding molecule also significantly reduced infection. Taken together, our findings support a role for TIM1 in eHEV-mediated cell entry, facilitated by the PS present on the viral membrane, a strategy HEV may use to promote viral spread throughout the infected body.
    MeSH term(s) Hepatitis E virus/genetics ; Hepatitis E virus/metabolism ; Virus Internalization ; Receptors, Cell Surface/metabolism ; Viruses
    Chemical Substances phosphatidylserine receptor ; Receptors, Cell Surface
    Language English
    Publishing date 2023-10-13
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 1358415-7
    ISSN 1420-9071 ; 1420-682X
    ISSN (online) 1420-9071
    ISSN 1420-682X
    DOI 10.1007/s00018-023-04977-4
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article: Hepatitis C virus alters the morphology and function of peroxisomes.

    Martin de Fourchambault, Esther / Callens, Nathalie / Saliou, Jean-Michel / Fourcot, Marie / Delos, Oceane / Barois, Nicolas / Thorel, Quentin / Ramirez, Santseharay / Bukh, Jens / Cocquerel, Laurence / Bertrand-Michel, Justine / Marot, Guillemette / Sebti, Yasmine / Dubuisson, Jean / Rouillé, Yves

    Frontiers in microbiology

    2023  Volume 14, Page(s) 1254728

    Abstract: Despite the introduction of effective treatments for hepatitis C in clinics, issues remain regarding the liver disease induced by chronic hepatitis C virus (HCV) infection. HCV is known to disturb the metabolism of infected cells, especially lipid ... ...

    Abstract Despite the introduction of effective treatments for hepatitis C in clinics, issues remain regarding the liver disease induced by chronic hepatitis C virus (HCV) infection. HCV is known to disturb the metabolism of infected cells, especially lipid metabolism and redox balance, but the mechanisms leading to HCV-induced pathogenesis are still poorly understood. In an APEX2-based proximity biotinylation screen, we identified ACBD5, a peroxisome membrane protein, as located in the vicinity of HCV replication complexes. Confocal microscopy confirmed the relocation of peroxisomes near HCV replication complexes and indicated that their morphology and number are altered in approximately 30% of infected Huh-7 cells. Peroxisomes are small versatile organelles involved among other functions in lipid metabolism and ROS regulation. To determine their importance in the HCV life cycle, we generated Huh-7 cells devoid of peroxisomes by inactivating the PEX5 and PEX3 genes using CRISPR/Cas9 and found that the absence of peroxisomes had no impact on replication kinetics or infectious titers of HCV strains JFH1 and DBN3a. The impact of HCV on peroxisomal functions was assessed using sub-genomic replicons. An increase of ROS was measured in peroxisomes of replicon-containing cells, correlated with a significant decrease of catalase activity with the DBN3a strain. In contrast, HCV replication had little to no impact on cytoplasmic and mitochondrial ROS, suggesting that the redox balance of peroxisomes is specifically impaired in cells replicating HCV. Our study provides evidence that peroxisome function and morphology are altered in HCV-infected cells.
    Language English
    Publishing date 2023-09-21
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2587354-4
    ISSN 1664-302X
    ISSN 1664-302X
    DOI 10.3389/fmicb.2023.1254728
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article: Processing and Subcellular Localization of the Hepatitis E Virus Replicase: Identification of Candidate Viral Factories.

    Metzger, Karoline / Bentaleb, Cyrine / Hervouet, Kévin / Alexandre, Virginie / Montpellier, Claire / Saliou, Jean-Michel / Ferrié, Martin / Camuzet, Charline / Rouillé, Yves / Lecoeur, Cécile / Dubuisson, Jean / Cocquerel, Laurence / Aliouat-Denis, Cécile-Marie

    Frontiers in microbiology

    2022  Volume 13, Page(s) 828636

    Abstract: Hepatitis E virus (HEV) is the major cause of acute hepatitis worldwide. HEV is a positive-sense RNA virus expressing three open reading frames (ORFs). ORF1 encodes the ORF1 non-structural polyprotein, the viral replicase which transcribes the full- ... ...

    Abstract Hepatitis E virus (HEV) is the major cause of acute hepatitis worldwide. HEV is a positive-sense RNA virus expressing three open reading frames (ORFs). ORF1 encodes the ORF1 non-structural polyprotein, the viral replicase which transcribes the full-length genome and a subgenomic RNA that encodes the structural ORF2 and ORF3 proteins. The present study is focused on the replication step with the aim to determine whether the ORF1 polyprotein is processed during the HEV lifecycle and to identify where the replication takes place inside the host cell. As no commercial antibody recognizes ORF1 in HEV-replicating cells, we aimed at inserting epitope tags within the ORF1 protein without impacting the virus replication efficacy. Two insertion sites located in the hypervariable region were thus selected to tolerate the V5 epitope while preserving HEV replication efficacy. Once integrated into the infectious full-length Kernow C-1 p6 strain, the V5 epitopes did neither impact the replication of genomic nor the production of subgenomic RNA. Also, the V5-tagged viral particles remained as infectious as the wildtype particles to Huh-7.5 cells. Next, the expression pattern of the V5-tagged ORF1 was compared in heterologous expression and replicative HEV systems. A high molecular weight protein (180 kDa) that was expressed in all three systems and that likely corresponds to the unprocessed form of ORF1 was detected up to 25 days after electroporation in the p6 cell culture system. Additionally, less abundant products of lower molecular weights were detected in both in cytoplasmic and nuclear compartments. Concurrently, the V5-tagged ORF1 was localized by confocal microscopy inside the cell nucleus but also as compact perinuclear substructures in which ORF2 and ORF3 proteins were detected. Importantly, using
    Language English
    Publishing date 2022-02-24
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2587354-4
    ISSN 1664-302X
    ISSN 1664-302X
    DOI 10.3389/fmicb.2022.828636
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  7. Article ; Online: CD81 and hepatitis C virus (HCV) infection.

    Fénéant, Lucie / Levy, Shoshana / Cocquerel, Laurence

    Viruses

    2014  Volume 6, Issue 2, Page(s) 535–572

    Abstract: Hepatitis C Virus (HCV) infection is a global public health problem affecting over 160 million individuals worldwide. Its symptoms include chronic hepatitis, liver cirrhosis and hepatocellular carcinoma. HCV is an enveloped RNA virus mainly targeting ... ...

    Abstract Hepatitis C Virus (HCV) infection is a global public health problem affecting over 160 million individuals worldwide. Its symptoms include chronic hepatitis, liver cirrhosis and hepatocellular carcinoma. HCV is an enveloped RNA virus mainly targeting liver cells and for which the initiation of infection occurs through a complex multistep process involving a series of specific cellular entry factors. This process is likely mediated through the formation of a tightly orchestrated complex of HCV entry factors at the plasma membrane. Among HCV entry factors, the tetraspanin CD81 is one of the best characterized and it is undoubtedly a key player in the HCV lifecycle. In this review, we detail the current knowledge on the involvement of CD81 in the HCV lifecycle, as well as in the immune response to HCV infection.
    MeSH term(s) Hepacivirus/physiology ; Host-Pathogen Interactions ; Humans ; Models, Biological ; Receptors, Virus/metabolism ; Tetraspanin 28/metabolism ; Virus Internalization
    Chemical Substances CD81 protein, human ; Receptors, Virus ; Tetraspanin 28
    Language English
    Publishing date 2014-02-06
    Publishing country Switzerland
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Review
    ZDB-ID 2516098-9
    ISSN 1999-4915 ; 1999-4915
    ISSN (online) 1999-4915
    ISSN 1999-4915
    DOI 10.3390/v6020535
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  8. Article ; Online: A ribavirin-induced ORF2 single-nucleotide variant produces defective hepatitis E virus particles with immune decoy function.

    Meister, Toni Luise / Brüggemann, Yannick / Nocke, Maximilian K / Ulrich, Rainer G / Schuhenn, Jonas / Sutter, Kathrin / Gömer, André / Bader, Verian / Winklhofer, Konstanze F / Broering, Ruth / Verhoye, Lieven / Meuleman, Philip / Vondran, Florian W R / Camuzet, Charline / Cocquerel, Laurence / Todt, Daniel / Steinmann, Eike

    Proceedings of the National Academy of Sciences of the United States of America

    2022  Volume 119, Issue 34, Page(s) e2202653119

    Abstract: Hepatitis E virus (HEV) is the causative agent of hepatitis E in humans and is the leading cause of enterically transmitted viral hepatitis worldwide. Ribavirin (RBV) is currently the only treatment option for many patients; however, cases of treatment ... ...

    Abstract Hepatitis E virus (HEV) is the causative agent of hepatitis E in humans and is the leading cause of enterically transmitted viral hepatitis worldwide. Ribavirin (RBV) is currently the only treatment option for many patients; however, cases of treatment failures or posttreatment relapses have been frequently reported. RBV therapy was shown to be associated with an increase in HEV genome heterogeneity and the emergence of distinct HEV variants. In this study, we analyzed the impact of eight patient-derived open reading frame 2 (ORF2) single-nucleotide variants (SNVs), which occurred under RBV treatment, on the replication cycle and pathogenesis of HEV. The parental HEV strain and seven ORF2 variants showed comparable levels of RNA replication in human hepatoma cells and primary human hepatocytes. However, a P79S ORF2 variant demonstrated reduced RNA copy numbers released in the supernatant and an impairment in the production of infectious particles. Biophysical and biochemical characterization revealed that this SNV caused defective, smaller HEV particles with a loss of infectiousness. Furthermore, the P79S variant displayed an altered subcellular distribution of the ORF2 protein and was able to interfere with antibody-mediated neutralization of HEV in a competition assay. In conclusion, an SNV in the HEV ORF2 could be identified that resulted in altered virus particles that were noninfectious in vitro and in vivo, but could potentially serve as immune decoys. These findings provide insights in understanding the biology of circulating HEV variants and may guide development of personalized antiviral strategies in the future.
    MeSH term(s) Cell Line, Tumor ; Hepatitis E virus/genetics ; Hepatitis E virus/physiology ; Hepatocytes/virology ; Humans ; Neoplasm Recurrence, Local/genetics ; Nucleotides ; RNA, Viral ; Ribavirin/pharmacology ; Viral Proteins/genetics ; Virus Replication
    Chemical Substances Nucleotides ; ORF2 protein, Hepatitis E virus ; RNA, Viral ; Viral Proteins ; Ribavirin (49717AWG6K)
    Language English
    Publishing date 2022-08-15
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 209104-5
    ISSN 1091-6490 ; 0027-8424
    ISSN (online) 1091-6490
    ISSN 0027-8424
    DOI 10.1073/pnas.2202653119
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  9. Article: Occludine, une clé de plus pour l'entrée du virus de l'hépatite C.

    Tews, Birke Andrea / Cocquerel, Laurence

    Medecine sciences : M/S

    2009  Volume 25, Issue 6-7, Page(s) 549–551

    Title translation Occludin, an additional key for hepatitis C virus entry.
    MeSH term(s) Hepacivirus/drug effects ; Hepatitis C/drug therapy ; Humans ; Membrane Proteins/physiology ; Membrane Proteins/therapeutic use ; Occludin ; Receptors, Virus/physiology
    Chemical Substances Membrane Proteins ; OCLN protein, human ; Occludin ; Receptors, Virus
    Language French
    Publishing date 2009-06
    Publishing country France
    Document type News
    ZDB-ID 632733-3
    ISSN 1958-5381 ; 0767-0974
    ISSN (online) 1958-5381
    ISSN 0767-0974
    DOI 10.1051/medsci/2009256-7549
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  10. Article ; Online: Hepatitis E Virus (HEV) Open Reading Frame 2 Antigen Kinetics in Human-Liver Chimeric Mice and Its Impact on HEV Diagnosis.

    Sayed, Ibrahim M / Verhoye, Lieven / Montpellier, Claire / Abravanel, Florence / Izopet, Jacques / Cocquerel, Laurence / Meuleman, Philip

    The Journal of infectious diseases

    2019  Volume 220, Issue 5, Page(s) 811–819

    Abstract: Background: Hepatitis E virus infection (HEV) is an emerging problem in developed countries. Diagnosis of HEV infection is based on the detection of HEV-specific antibodies, viral RNA, and/or antigen (Ag). Humanized mice were previously reported as a ... ...

    Abstract Background: Hepatitis E virus infection (HEV) is an emerging problem in developed countries. Diagnosis of HEV infection is based on the detection of HEV-specific antibodies, viral RNA, and/or antigen (Ag). Humanized mice were previously reported as a model for the study of HEV infection, but published data were focused on the quantification of viral RNA. However, the kinetics of HEV Ag expression during infection remains poorly understood.
    Methods: Plasma specimens and suspensions of fecal specimens from HEV-infected and ribavirin-treated humanized mice were analyzed using HEV antigen-specific enzyme-linked immunosorbent assay, reverse transcription-quantitative polymerase chain reaction analysis, density gradient analysis, and Western blotting.
    Result: Open reading frame 2 (ORF2) Ag was detected in both plasma and stool from HEV-infected mice, and levels increased over time. Contrary to HEV RNA, ORF2 Ag levels were higher in mouse plasma than in stool. Interestingly, ORF2 was detected in plasma from mice that tested negative for HEV RNA in plasma but positive for HEV RNA in stool and was detected after viral clearance in mice that were treated with ribavirin. Plasma density gradient analysis revealed the presence of the noninfectious glycosylated form of ORF2.
    Conclusion: ORF2 Ag can be used as a marker of active HEV infection and for assessment of the effect of antiviral therapy, especially when fecal samples are not available or molecular diagnostic tests are not accessible.
    MeSH term(s) Animals ; Disease Models, Animal ; Feces/virology ; Hepatitis Antigens/immunology ; Hepatitis E/diagnosis ; Hepatitis E/drug therapy ; Hepatitis E/immunology ; Hepatitis E virus/drug effects ; Hepatitis E virus/immunology ; Humans ; Kinetics ; Liver/virology ; Mice ; Mice, SCID ; RNA, Viral/isolation & purification ; Ribavirin ; Viral Proteins/immunology
    Chemical Substances Hepatitis Antigens ; ORF2 protein, Hepatitis E virus ; RNA, Viral ; Viral Proteins ; Ribavirin (49717AWG6K)
    Language English
    Publishing date 2019-03-27
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 3019-3
    ISSN 1537-6613 ; 0022-1899
    ISSN (online) 1537-6613
    ISSN 0022-1899
    DOI 10.1093/infdis/jiz171
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