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  1. Article ; Online: Analysis of the 5'UTR of HCV genotype 3 grown in vitro in human B cells, T cells, and macrophages

    Prichard John G / Alberti Michael O / Revie Dennis / Kelley Ann S / Salahuddin S Zaki

    Virology Journal, Vol 7, Iss 1, p

    2010  Volume 155

    Abstract: Abstract Background Previously, we have reported the isolation and molecular characterization of human Hepatitis C virus genotype 1 (HCV-1) from infected patients. We are now reporting an analysis of HCV obtained from patients infected with HCV genotype ... ...

    Abstract Abstract Background Previously, we have reported the isolation and molecular characterization of human Hepatitis C virus genotype 1 (HCV-1) from infected patients. We are now reporting an analysis of HCV obtained from patients infected with HCV genotype 3 (HCV-3) as diagnosed by clinical laboratories. Results HCV was cultured in vitro using our system. HCV RNA was isolated from patients' blood and from HCV cultured in various cell types for up to three months. The 5'UTR of these isolates were used for comparisons. Results revealed a number of sequence changes as compared to the serum RNA. The HCV RNA produced efficiently by infected macrophages, B-cells, and T-cells had sequences similar to HCV-1, which suggests that selection of the variants was performed at the level of macrophages. Virus with sequences similar to HCV-1 replicated better in macrophages than HCV having a 5'UTR similar to HCV-3. Conclusions Although HCV-3 replicates in cell types such as B-cells, T-cells, and macrophages, it may require a different primary cell type for the same purpose. Therefore, in our opinion, HCV-3 does not replicate efficiently in macrophages, and patients infected with HCV-3 may contain a population of HCV-1 in their blood.
    Keywords Microbiology ; QR1-502 ; Science ; Q ; DOAJ:Microbiology ; DOAJ:Biology ; DOAJ:Biology and Life Sciences ; Medicine (General) ; R5-920 ; Medicine ; R ; DOAJ:Medicine (General) ; DOAJ:Health Sciences
    Subject code 360
    Language English
    Publishing date 2010-07-01T00:00:00Z
    Publisher BioMed Central
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  2. Article ; Online: Analysis of the 5'UTR of HCV genotype 3 grown in vitro in human B cells, T cells, and macrophages

    Revie, Dennis / Alberti, Michael O / Prichard, John G / Kelley, Ann S / Salahuddin, S. Zaki

    Virology journal. 2010 Dec., v. 7, no. 1 p.155-155

    2010  

    Abstract: BACKGROUND: Previously, we have reported the isolation and molecular characterization of human Hepatitis C virus genotype 1 (HCV-1) from infected patients. We are now reporting an analysis of HCV obtained from patients infected with HCV genotype 3 (HCV-3) ...

    Abstract BACKGROUND: Previously, we have reported the isolation and molecular characterization of human Hepatitis C virus genotype 1 (HCV-1) from infected patients. We are now reporting an analysis of HCV obtained from patients infected with HCV genotype 3 (HCV-3) as diagnosed by clinical laboratories. RESULTS: HCV was cultured in vitro using our system. HCV RNA was isolated from patients' blood and from HCV cultured in various cell types for up to three months. The 5'UTR of these isolates were used for comparisons. Results revealed a number of sequence changes as compared to the serum RNA. The HCV RNA produced efficiently by infected macrophages, B-cells, and T-cells had sequences similar to HCV-1, which suggests that selection of the variants was performed at the level of macrophages. Virus with sequences similar to HCV-1 replicated better in macrophages than HCV having a 5'UTR similar to HCV-3. CONCLUSIONS: Although HCV-3 replicates in cell types such as B-cells, T-cells, and macrophages, it may require a different primary cell type for the same purpose. Therefore, in our opinion, HCV-3 does not replicate efficiently in macrophages, and patients infected with HCV-3 may contain a population of HCV-1 in their blood.
    Keywords 5' untranslated regions ; B-lymphocytes ; Hepatitis C virus ; T-lymphocytes ; genotype ; human cell lines ; macrophages ; virus replication
    Language English
    Dates of publication 2010-12
    Size p. 155.
    Publishing place BioMed Central
    Document type Article ; Online
    ZDB-ID 2160640-7
    ISSN 1743-422X
    ISSN 1743-422X
    DOI 10.1186/1743-422X-7-155
    Database NAL-Catalogue (AGRICOLA)

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  3. Article ; Online: Analysis of the 5'UTR of HCV genotype 3 grown in vitro in human B cells, T cells, and macrophages.

    Revie, Dennis / Alberti, Michael O / Prichard, John G / Kelley, Ann S / Salahuddin, S Zaki

    Virology journal

    2010  Volume 7, Page(s) 155

    Abstract: Background: Previously, we have reported the isolation and molecular characterization of human Hepatitis C virus genotype 1 (HCV-1) from infected patients. We are now reporting an analysis of HCV obtained from patients infected with HCV genotype 3 (HCV- ... ...

    Abstract Background: Previously, we have reported the isolation and molecular characterization of human Hepatitis C virus genotype 1 (HCV-1) from infected patients. We are now reporting an analysis of HCV obtained from patients infected with HCV genotype 3 (HCV-3) as diagnosed by clinical laboratories.
    Results: HCV was cultured in vitro using our system. HCV RNA was isolated from patients' blood and from HCV cultured in various cell types for up to three months. The 5'UTR of these isolates were used for comparisons. Results revealed a number of sequence changes as compared to the serum RNA. The HCV RNA produced efficiently by infected macrophages, B-cells, and T-cells had sequences similar to HCV-1, which suggests that selection of the variants was performed at the level of macrophages. Virus with sequences similar to HCV-1 replicated better in macrophages than HCV having a 5'UTR similar to HCV-3.
    Conclusions: Although HCV-3 replicates in cell types such as B-cells, T-cells, and macrophages, it may require a different primary cell type for the same purpose. Therefore, in our opinion, HCV-3 does not replicate efficiently in macrophages, and patients infected with HCV-3 may contain a population of HCV-1 in their blood.
    MeSH term(s) 5' Untranslated Regions ; B-Lymphocytes/virology ; Base Sequence ; Genotype ; Hepacivirus/classification ; Hepacivirus/genetics ; Hepacivirus/growth & development ; Hepacivirus/isolation & purification ; Hepatitis C/virology ; Humans ; Macrophages/virology ; Molecular Sequence Data ; Phylogeny ; Sequence Alignment ; T-Lymphocytes/virology ; Virus Replication
    Chemical Substances 5' Untranslated Regions
    Language English
    Publishing date 2010-07-13
    Publishing country England
    Document type Journal Article
    ISSN 1743-422X
    ISSN (online) 1743-422X
    DOI 10.1186/1743-422X-7-155
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: The simultaneous presence and expression of human hepatitis C virus (HCV), human herpesvirus-6 (HHV-6), and human immunodeficiency virus-1 (HIV-1) in a single human T-cell

    Prichard John G / Grewal Renu / Godwin Andre / Snyder Katherine A / Salahuddin S Zaki / Kelley Ann S / Revie Dennis

    Virology Journal, Vol 4, Iss 1, p

    2007  Volume 106

    Abstract: Abstract We have developed a system that isolates and replicates HCV in vitro . These isolates are called CIMM-HCV. This system has made it possible to analyze the biology, nature, and extent of HCV variability, among other things. Individuals that are ... ...

    Abstract Abstract We have developed a system that isolates and replicates HCV in vitro . These isolates are called CIMM-HCV. This system has made it possible to analyze the biology, nature, and extent of HCV variability, among other things. Individuals that are infected with HIV-1 are often also infected with HCV and HHV-6. In addition to HCV, our lab has systems for replicating HIV-1 and HHV-6. We asked whether all these viruses could infect the same cells. We report here the successful infection of a T-cell (CEM) by CIMM-HCV, HHV-6, and HIV-1. PCR analyses demonstrated that the CEM cells were productively infected by HHV-6A. RT-PCR showed that the same cell culture was positive for HCV and HIV-1. Co-infection of a T-cell by all three viruses was confirmed by transmission electron microscopy (TEM). All these viruses are highly cytolytic; therefore, triply-infected cells were short lived. However, HIV-1 and HCV co-infected cells unexpectedly lasted for several weeks. Viral replication was unhindered and the phenomenon of 'dominance' was not observed in our experiments. In addition, CIMM-HCV was present in the perinuclear space, suggesting their possible synthesis in the nucleus. This report is based entirely on viruses produced in vitro in our laboratories. As part of the determinations of host ranges of these viruses, studies were designed to demonstrate the infection of a single cell by these viruses and to study the consequences of this phenomenon. All measurements were made on cultured cells and cell culture supernatants.
    Keywords Microbiology ; QR1-502 ; Science ; Q ; DOAJ:Microbiology ; DOAJ:Biology ; DOAJ:Biology and Life Sciences ; Medicine (General) ; R5-920 ; Medicine ; R ; DOAJ:Medicine (General) ; DOAJ:Health Sciences
    Subject code 570
    Language English
    Publishing date 2007-10-01T00:00:00Z
    Publisher BioMed Central
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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    Inter-library loan at ZB MED

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  5. Article ; Online: The simultaneous presence and expression of human hepatitis C virus (HCV), human herpesvirus-6 (HHV-6), and human immunodeficiency virus-1 (HIV-1) in a single human T-cell.

    Salahuddin, S Zaki / Snyder, Katherine A / Godwin, Andre / Grewal, Renu / Prichard, John G / Kelley, Ann S / Revie, Dennis

    Virology journal

    2007  Volume 4, Page(s) 106

    Abstract: We have developed a system that isolates and replicates HCV in vitro. These isolates are called CIMM-HCV. This system has made it possible to analyze the biology, nature, and extent of HCV variability, among other things. Individuals that are infected ... ...

    Abstract We have developed a system that isolates and replicates HCV in vitro. These isolates are called CIMM-HCV. This system has made it possible to analyze the biology, nature, and extent of HCV variability, among other things. Individuals that are infected with HIV-1 are often also infected with HCV and HHV-6. In addition to HCV, our lab has systems for replicating HIV-1 and HHV-6. We asked whether all these viruses could infect the same cells. We report here the successful infection of a T-cell (CEM) by CIMM-HCV, HHV-6, and HIV-1. PCR analyses demonstrated that the CEM cells were productively infected by HHV-6A. RT-PCR showed that the same cell culture was positive for HCV and HIV-1. Co-infection of a T-cell by all three viruses was confirmed by transmission electron microscopy (TEM). All these viruses are highly cytolytic; therefore, triply-infected cells were short lived. However, HIV-1 and HCV co-infected cells unexpectedly lasted for several weeks. Viral replication was unhindered and the phenomenon of 'dominance' was not observed in our experiments. In addition, CIMM-HCV was present in the perinuclear space, suggesting their possible synthesis in the nucleus. This report is based entirely on viruses produced in vitro in our laboratories. As part of the determinations of host ranges of these viruses, studies were designed to demonstrate the infection of a single cell by these viruses and to study the consequences of this phenomenon. All measurements were made on cultured cells and cell culture supernatants.
    MeSH term(s) Cell Culture Techniques ; HIV-1/genetics ; HIV-1/physiology ; Hepacivirus/genetics ; Hepacivirus/physiology ; Herpesvirus 6, Human/genetics ; Herpesvirus 6, Human/physiology ; Humans ; Microscopy, Electron, Transmission ; T-Lymphocytes/ultrastructure ; T-Lymphocytes/virology ; Virion/growth & development ; Virus Replication/genetics
    Language English
    Publishing date 2007-10-24
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 1743-422X
    ISSN (online) 1743-422X
    DOI 10.1186/1743-422X-4-106
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article ; Online: Transmission of human hepatitis C virus from patients in secondary cells for long term culture

    Geer Cheryl / Khan Rafat / Chelyapov Nickolas / Bayles David / Braich Ravi S / Revie Dennis / Reisman Richard / Kelley Ann S / Prichard John G / Salahuddin S Zaki

    Virology Journal, Vol 2, Iss 1, p

    2005  Volume 37

    Abstract: Abstract Infection by human hepatitis C virus (HCV) is the principal cause of post-transfusion hepatitis and chronic liver diseases worldwide. A reliable in vitro culture system for the isolation and analysis of this virus is not currently available, and, ...

    Abstract Abstract Infection by human hepatitis C virus (HCV) is the principal cause of post-transfusion hepatitis and chronic liver diseases worldwide. A reliable in vitro culture system for the isolation and analysis of this virus is not currently available, and, as a consequence, HCV pathogenesis is poorly understood. We report here the first robust in vitro system for the isolation and propagation of HCV from infected donor blood. This system involves infecting freshly prepared macrophages with HCV and then transmission of macrophage-adapted virus into freshly immortalized B-cells from human fetal cord blood. Using this system, newly isolated HCV have been replicated in vitro in continuous cultures for over 130 weeks. These isolates were also transmitted by cell-free methods into different cell types, including B-cells, T-cells and neuronal precursor cells. These secondarily infected cells also produced in vitro transmissible infectious virus. Replication of HCV-RNA was validated by RT-PCR analysis and by in situ hybridization. Although nucleic acid sequencing of the HCV isolate reported here indicates that the isolate is probably of type 1a, other HCV types have also been isolated using this system. Western blot analysis shows the synthesis of major HCV structural proteins. We present here, for the first time, a method for productively growing HCV in vitro for prolonged periods of time. This method allows studies related to understanding the replication process, viral pathogenesis, and the development of anti-HCV drugs and vaccines.
    Keywords Microbiology ; QR1-502 ; Science ; Q ; DOAJ:Microbiology ; DOAJ:Biology ; DOAJ:Biology and Life Sciences ; Medicine (General) ; R5-920 ; Medicine ; R ; DOAJ:Medicine (General) ; DOAJ:Health Sciences
    Subject code 570
    Language English
    Publishing date 2005-04-01T00:00:00Z
    Publisher BioMed Central
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

    Full text online

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    Inter-library loan at ZB MED

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  7. Article ; Online: Transmission of human hepatitis C virus from patients in secondary cells for long term culture.

    Revie, Dennis / Braich, Ravi S / Bayles, David / Chelyapov, Nickolas / Khan, Rafat / Geer, Cheryl / Reisman, Richard / Kelley, Ann S / Prichard, John G / Salahuddin, S Zaki

    Virology journal

    2005  Volume 2, Page(s) 37

    Abstract: Infection by human hepatitis C virus (HCV) is the principal cause of post-transfusion hepatitis and chronic liver diseases worldwide. A reliable in vitro culture system for the isolation and analysis of this virus is not currently available, and, as a ... ...

    Abstract Infection by human hepatitis C virus (HCV) is the principal cause of post-transfusion hepatitis and chronic liver diseases worldwide. A reliable in vitro culture system for the isolation and analysis of this virus is not currently available, and, as a consequence, HCV pathogenesis is poorly understood. We report here the first robust in vitro system for the isolation and propagation of HCV from infected donor blood. This system involves infecting freshly prepared macrophages with HCV and then transmission of macrophage-adapted virus into freshly immortalized B-cells from human fetal cord blood. Using this system, newly isolated HCV have been replicated in vitro in continuous cultures for over 130 weeks. These isolates were also transmitted by cell-free methods into different cell types, including B-cells, T-cells and neuronal precursor cells. These secondarily infected cells also produced in vitro transmissible infectious virus. Replication of HCV-RNA was validated by RT-PCR analysis and by in situ hybridization. Although nucleic acid sequencing of the HCV isolate reported here indicates that the isolate is probably of type 1a, other HCV types have also been isolated using this system. Western blot analysis shows the synthesis of major HCV structural proteins. We present here, for the first time, a method for productively growing HCV in vitro for prolonged periods of time. This method allows studies related to understanding the replication process, viral pathogenesis, and the development of anti-HCV drugs and vaccines.
    MeSH term(s) B-Lymphocytes/virology ; Cells, Cultured ; Hepacivirus/physiology ; Hepatitis C/virology ; Humans ; Macrophages/virology ; Neurons/virology ; RNA, Viral/metabolism ; Stem Cells/virology ; T-Lymphocytes/virology ; Virus Cultivation/methods ; Virus Replication
    Chemical Substances RNA, Viral
    Language English
    Publishing date 2005-04-19
    Publishing country England
    Document type Journal Article ; Research Support, U.S. Gov't, Non-P.H.S.
    ZDB-ID 2160640-7
    ISSN 1743-422X ; 1743-422X
    ISSN (online) 1743-422X
    ISSN 1743-422X
    DOI 10.1186/1743-422X-2-37
    Database MEDical Literature Analysis and Retrieval System OnLINE

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