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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 SZ, Snyder KA, Godwin A, Grewal R, Prichard JG, Kelley AS, Revie D - Virol. J. (2007)

Bottom Line: We report here the successful infection of a T-cell (CEM) by CIMM-HCV, HHV-6, and HIV-1.In addition, CIMM-HCV was present in the perinuclear space, suggesting their possible synthesis in the nucleus.All measurements were made on cultured cells and cell culture supernatants.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Basic Research, California Institute of Molecular Medicine, Ventura, California, USA. zaki@cimm.net

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.

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Cytopathic effects in CEM cells. This picture represents the commonly observed cytopathic effects in triply-infected CEM cells. Large cells are seen with increasing frequency in infected cultures.
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Figure 2: Cytopathic effects in CEM cells. This picture represents the commonly observed cytopathic effects in triply-infected CEM cells. Large cells are seen with increasing frequency in infected cultures.

Mentions: CEM cells were infected individually by each of the three viruses (Fig. 1). To test the infectivity of HHV-6A, CEM cells were infected with the virus (Figure 1, K1) [9,10]. Two methods were used to determine whether the cell culture was infected and actively producing HHV-6A particles. First, characteristic cytopathic effects (CPE) on the cells were observed. The cells have the appearance of a balloon, called "juicy cells" (Figure 2) [10]. Second, the cell culture supernatants were tested by PCR using the appropriate primer set (Table 2). A band of about 400 bp was seen after PCR analysis, indicating HHV-6A was replicating in these cells (Figure 3A, Lane 3).


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 SZ, Snyder KA, Godwin A, Grewal R, Prichard JG, Kelley AS, Revie D - Virol. J. (2007)

Cytopathic effects in CEM cells. This picture represents the commonly observed cytopathic effects in triply-infected CEM cells. Large cells are seen with increasing frequency in infected cultures.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2170440&req=5

Figure 2: Cytopathic effects in CEM cells. This picture represents the commonly observed cytopathic effects in triply-infected CEM cells. Large cells are seen with increasing frequency in infected cultures.
Mentions: CEM cells were infected individually by each of the three viruses (Fig. 1). To test the infectivity of HHV-6A, CEM cells were infected with the virus (Figure 1, K1) [9,10]. Two methods were used to determine whether the cell culture was infected and actively producing HHV-6A particles. First, characteristic cytopathic effects (CPE) on the cells were observed. The cells have the appearance of a balloon, called "juicy cells" (Figure 2) [10]. Second, the cell culture supernatants were tested by PCR using the appropriate primer set (Table 2). A band of about 400 bp was seen after PCR analysis, indicating HHV-6A was replicating in these cells (Figure 3A, Lane 3).

Bottom Line: We report here the successful infection of a T-cell (CEM) by CIMM-HCV, HHV-6, and HIV-1.In addition, CIMM-HCV was present in the perinuclear space, suggesting their possible synthesis in the nucleus.All measurements were made on cultured cells and cell culture supernatants.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Basic Research, California Institute of Molecular Medicine, Ventura, California, USA. zaki@cimm.net

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.

Show MeSH
Related in: MedlinePlus