Isolation of a heterogeneous population of temperature-sensitive mutants of measles virus from persistently infected human lymphoblastoid cell lines.

Two human lymphoblastoid B-cell lines, WI-L2 and 8866, were infected with the Edmonston strain of measles virus at a multiplicity of infection of 10(-6), and stable persistent infections were established. By immunofluorescence and electron microscopy, the vast majority of cells from both cell lines were expressing viral antigens and releasing virion-like particles. However, very little infectious virus could be detected at 37 degrees C, either by an infectious centers assay or by titration of supernates from persistently infected cultures. When cultures were shifted to 31 degrees C, the cells released a population of virus that was temperature-sensitive. Clonal analysis of supernatant virus at 31 degrees C revealed a highly heterogeneous population of temperature-sensitive mutants, differing in plating efficiency ratios, thermolability, and antigen production at the nonpermissive temperature. Factors such as interferon, defective interfering particles, and extracellular virus do not appear to be important in maintaining the persistent carrier state. These studies have important implications for persistent infections of lymphoid cells in vivo, and the slow neurological diseases associated with measles, subacute sclerosing panencephalitis, and multiple sclerosis.

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Temperature-sensitive mutants isolated from hamster and canine cell lines persistently infected with Newcastle disease virus.

Journal of Virology ◽

10.1128/jvi.16.5.1332-1336.1975 ◽

1975 ◽

Vol 16 (5) ◽

pp. 1332-1336 ◽

Cited By ~ 13

Author(s):

J S Youngner ◽

D O Quagliana

Keyword(s):

Newcastle Disease Virus ◽

Disease Virus ◽

Cell Lines ◽

Newcastle Disease ◽

Temperature Sensitive ◽

Persistently Infected ◽

Temperature Sensitive Mutants

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Temperature-sensitive mutants of measles virus produced from persistently infected HeLa cells

Archives of Virology ◽

10.1007/bf01314853 ◽

1977 ◽

Vol 53 (1-2) ◽

pp. 121-132 ◽

Cited By ~ 12

Author(s):

R. C. Armen ◽

J. F. Evermann ◽

A. L. Truant ◽

C. A. Laughlin ◽

J. V. Hallum

Keyword(s):

Hela Cells ◽

Measles Virus ◽

Temperature Sensitive ◽

Persistently Infected ◽

Temperature Sensitive Mutants

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Type I Interferon acts as a major barrier to the establishment of infectious bursal disease virus (IBDV) persistent infections

Journal of Virology ◽

10.1128/jvi.02017-20 ◽

2020 ◽

pp. JVI.02017-20

Author(s):

Laura Broto ◽

Nicolás Romero ◽

Fernando Méndez ◽

Elisabet Diaz-Beneitez ◽

Oscar Candelas-Rivera ◽

...

Keyword(s):

Disease Virus ◽

Cell Lines ◽

Infectious Bursal Disease Virus ◽

Infectious Bursal Disease ◽

Type I ◽

Persistently Infected ◽

Persistent Infections ◽

Avian Pathogen ◽

Bursal Disease ◽

Infected Cells

Infectious bursal disease virus (IBDV), the best characterized member of the Birnaviridae family, is a highly relevant avian pathogen causing both acute and persistent infections in different avian hosts. Here, we describe the establishment of clonal, long-term, productive persistent IBDV infections in DF-1 chicken embryonic fibroblasts. Although virus yields in persistently-infected cells are exceedingly lower than those detected in acutely infected cells, the replication fitness of viruses isolated from persistently-infected cells is higher than that of the parental virus. Persistently-infected DF-1 and IBDV-cured cell lines derived from them do not respond to type I interferon (IFN). High-throughput genome sequencing revealed that this defect is due to mutations affecting the IFNα/β receptor subunit 2 (IFNAR2) gene resulting in the expression of IFNAR2 polypeptides harbouring large C-terminal deletions that abolish the signalling capacity of IFNα/β receptor complex. Ectopic expression of a recombinant chicken IFNAR2 gene efficiently rescues IFNα responsiveness. IBDV-cured cell lines derived from persistently infected cells exhibit a drastically enhanced susceptibility to establishing new persistent IBDV infections. Additionally, experiments carried out with human HeLa cells lacking the IFNAR2 gene fully recapitulate results obtained with DF-1 cells, exhibiting a highly enhanced capacity to both survive the acute IBDV infection phase and to support the establishment of persistent IBDV infections. Results presented here show that the inactivation of the JAK-STAT signalling pathway significantly reduces the apoptotic response induced by the infection, hence facilitating the establishment and maintenance of IBDV persistent infections.IMPORTANCE Members of the Birnaviridae family, including infectious bursal disease virus (IBDV), exhibit a dual behaviour, causing acute infections that are often followed by the establishment of life-long persistent asymptomatic infections. Indeed, persistently infected specimens might act as efficient virus reservoirs, hence potentially contributing to virus dissemination. Despite the key importance of this biological trait, information about mechanisms triggering IBDV persistency is negligible. Our report evidences the capacity of IBDV, a highly relevant avian pathogen, to establishing long-term, productive, persistent infections in both avian and human cell lines. Data presented here provide novel and direct evidence about the crucial role of type I IFNs on the fate of IBDV-infected cells and their contribution to controlling the establishment of IBDV persistent infections. The use of cell lines unable to respond to type I IFNs opens a promising venue to unveiling additional factors contributing to IBDV persistency.

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Type I Interferon acts as a major barrier to the establishment of infectious bursal disease virus (IBDV) persistent infections

10.1101/2020.10.09.333161 ◽

2020 ◽

Author(s):

Laura Broto ◽

Nicolás Romero ◽

Fernando Méndez ◽

Elisabet Diaz-Beneitez ◽

Oscar Candelas-Rivera ◽

...

Keyword(s):

Disease Virus ◽

Cell Lines ◽

Infectious Bursal Disease Virus ◽

Infectious Bursal Disease ◽

Type I ◽

Persistently Infected ◽

Persistent Infections ◽

Avian Pathogen ◽

Bursal Disease ◽

Infected Cells

ABSTRACTInfectious bursal disease virus (IBDV), the best characterized member of the Birnaviridae family, is a highly relevant avian pathogen causing both acute and persistent infections in different avian hosts. Here, we describe the establishment of clonal, long-term, productive persistent IBDV infections in DF-1 chicken embryonic fibroblasts. Although virus yields in persistently-infected cells are exceedingly lower than those detected in acutely infected cells, the replication fitness of viruses isolated from persistently-infected cells is higher than that of the parental virus. Persistently-infected DF-1 and IBDV-cured cell lines derived from them do not respond to type I interferon (IFN). High-throughput genome sequencing revealed that this defect is due to mutations affecting the IFNα/β receptor subunit 2 (IFNAR2) gene resulting in the expression of IFNAR2 polypeptides harbouring large C-terminal deletions that abolish the signalling capacity of IFNα/β receptor complex. Ectopic expression of a recombinant chicken IFNAR2 gene efficiently rescues IFNα responsiveness. IBDV-cured cell lines derived from persistently infected cells exhibit a drastically enhanced proneness to establishing new persistent IBDV infections. Additionally, experiments carried out with human HeLa cells lacking the IFNAR2 gene fully recapitulate results obtained with DF-1 cells, exhibiting a highly enhanced capacity to both survive the acute IBDV infection phase and to support the establishment of persistent IBDV infections. Results presented here show that the inactivation of the JAK-STAT signalling pathway significantly reduces the apoptotic response induced by the infection, hence facilitating the establishment and maintenance of IBDV persistent infections.IMPORTANCEMembers of the Birnaviridae family, including infectious bursal disease virus (IBDV), exhibit a dual behaviour, causing acute infections that are often followed by the establishment of life-long persistent asymptomatic infections. Indeed, persistently infected specimens might act as efficient virus reservoirs, hence potentially contributing to virus dissemination. Despite the key importance of this biological trait, information about mechanisms triggering IBDV persistency is negligible. Our report evidences the capacity of IBDV, a highly relevant avian pathogen, to establishing long-term, productive, persistent infections in both avian and human cell lines. Data presented here provide novel and direct evidence about the crucial role of type I IFNs on the fate of IBDV-infected cells and their contribution to controlling the establishment of IBDV persistent infections. The use of cell lines unable to respond to type I IFNs opens a promising venue to unveiling additional factors contributing to IBDV persistency.

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