Clearance of Measles Virus from Persistently Infected Cells by Short Hairpin RNA

Zinke, Michael; Kendl, Sabine; Singethan, Katrin; Fehrholz, Markus; Reuter, Dajana; Rennick, Linda J.; Herold, Marco J.; Schneider-Schaulies, J.

doi:10.1128/jvi.00846-09

Cited by 12 publications

(15 citation statements)

References 41 publications

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“…However, this model has not been further developed or used in recent years. Instead, an in vitro culture system of different CNS cell types is being increasingly used to study the mechanisms involved in morbillivirus CNS persistence and to explore therapeutic strategies [67,68,69]. …”

Section: Neurologic Complicationsmentioning

confidence: 99%

Morbillivirus Experimental Animal Models: Measles Virus Pathogenesis Insights from Canine Distemper Virus

Budaszewski

Messling

2016

Viruses

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Morbilliviruses share considerable structural and functional similarities. Even though disease severity varies among the respective host species, the underlying pathogenesis and the clinical signs are comparable. Thus, insights gained with one morbillivirus often apply to the other members of the genus. Since the Canine distemper virus (CDV) causes severe and often lethal disease in dogs and ferrets, it is an attractive model to characterize morbillivirus pathogenesis mechanisms and to evaluate the efficacy of new prophylactic and therapeutic approaches. This review compares the cellular tropism, pathogenesis, mechanisms of persistence and immunosuppression of the Measles virus (MeV) and CDV. It then summarizes the contributions made by studies on the CDV in dogs and ferrets to our understanding of MeV pathogenesis and to vaccine and drugs development.

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Section: Neurologic Complicationsmentioning

confidence: 99%

Morbillivirus Experimental Animal Models: Measles Virus Pathogenesis Insights from Canine Distemper Virus

Budaszewski

Messling

2016

Viruses

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“…RNA interference has been described to be active against MV infections [70,130,131] and is principally applicable against the intracellularly replicating viral RNPs in SSPE brains. In our own studies [70,133], we found that silencing of the viral RNP-mRNAs was highly efficient in reducing viral messenger and genomic RNA, whereas siRNA-mediated knockdown of the M protein not only enhanced cell-cell fusion, but also increased the levels of both, viral messenger and genomic RNAs by a factor of 2 to 2.5. These findings support older findings that the M protein acts as a negative regulator of the viral polymerase [132], a mechanism that probably contributes to the efficient replication and spread of M-defective RNPs in SSPE brains.…”

Section: Approaches To a Specific Therapymentioning

confidence: 93%

“…These findings support older findings that the M protein acts as a negative regulator of the viral polymerase [132], a mechanism that probably contributes to the efficient replication and spread of M-defective RNPs in SSPE brains. Recently, the efficacy of lentiviral vector-constructs, which integrate after transduction into the cellular genome and permanently express antiviral short hairpin RNAs (shRNA), was tested using persistently MV-infected NT2 cells (human teratocarcinoma cells that can be terminally differentiated into neuronal cells by retinoic acid treatment) [133]. ShRNA against N, P and L -mRNAs led to a reduction of viral proteins below detectable levels in a high percentage of transduced cells and complete virus elimination within 2 weeks [133].…”

Section: Approaches To a Specific Therapymentioning

confidence: 99%

“…Recently, the efficacy of lentiviral vector-constructs, which integrate after transduction into the cellular genome and permanently express antiviral short hairpin RNAs (shRNA), was tested using persistently MV-infected NT2 cells (human teratocarcinoma cells that can be terminally differentiated into neuronal cells by retinoic acid treatment) [133]. ShRNA against N, P and L -mRNAs led to a reduction of viral proteins below detectable levels in a high percentage of transduced cells and complete virus elimination within 2 weeks [133]. This demonstrated in tissue culture that persistently MV-infected neural cells can be cured by transduction of lentiviral vectors mediating long lasting expression of virus-specific shRNA.…”

Section: Approaches To a Specific Therapymentioning

confidence: 99%

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Measles virus infection of the CNS: human disease, animal models, and approaches to therapy

Reuter

Schneider-Schaulies

2010

Med Microbiol Immunol

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Viral infections of the central nervous system(CNS) mostly represent clinically important, often life-threatening complications of systemic viral infections. After acute measles, CNS complications may occur early (acute postinfectious measles encephalitis, APME) or after years of viral persistence (subacute sclerosing panencephalitis, SSPE). In spite of a presumably functional cell-mediated immunity and high antiviral antibody titers, an immunological control of the CNS infection is not achieved in patients suffering from SSPE. There is still no specific therapy for acute complications and persistent MV infections of the CNS. Hamsters, rats, and (genetically unmodified and modified) mice have been used as model systems to study mechanisms of MV-induced CNS infections. Functional CD4+ and CD8+ T cells together with IFN-gamma are required to overcome the infection. With the help of recombinant measles viruses and mice expressing endogenous or transgenic receptors, interesting aspects such as receptor-dependent viral spread and viral determinants of virulence have been investigated. However, many questions concerning the lack of efficient immune control in the CNS are still open. Recent research opened new perspectives using specific antivirals such as short interfering RNA (siRNA) or small molecule inhibitors. Inspite of obvious hurdles, these treatments are the most promising approaches to future therapies.

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“…Cloning of lentiviral expression plasmids and production of pseudotyped particles. For shRNA expression, we used the vector F6gW-DsRed, also expressing DsRed2 as a control for transduction efficiency, as described previously (57). For cloning of shRNA-expressing vectors, we selected three siRNA sequences from published mRNA sequences using the program provided by Block-iT RNAi designer (Invitrogen) and used DNA oligonucleotides for cloning into pF6gW-DsRed (the sequences of all primers used can be provided on request).…”

Section: Cells and Virusesmentioning

confidence: 99%

APOBEC3G-Regulated Host Factors Interfere with Measles Virus Replication: Role of REDD1 and Mammalian TORC1 Inhibition

Tiwarekar

Wohlfahrt

Fehrholz

et al. 2018

J Virol

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We found earlier that ectopic expression of the cytidine deaminase APOBEC3G (A3G) in Vero cells inhibits measles virus (MV), respiratory syncytial virus, and mumps virus, while the mechanism of inhibition remained unclear. A microarray analysis revealed that in A3G-transduced Vero cells, several cellular transcripts were differentially expressed, suggesting that A3G regulates the expression of host factors. One of the most upregulated host cell factors, REDD1 (regulated in development and DNA damage response-1, also called DDIT4), reduced MV replication ∼10-fold upon overexpression in Vero cells. REDD1 is an endogenous inhibitor of mTORC1 (mammalian target of rapamycin complex-1), the central regulator of cellular metabolism. Interestingly, rapamycin reduced the MV replication similarly to REDD1 overexpression, while the combination of both did not lead to further inhibition, suggesting that the same pathway is affected. REDD1 silencing in A3G-expressing Vero cells abolished the inhibitory effect of A3G. In addition, silencing of A3G led to reduced REDD1 expression, confirming that its expression is regulated by A3G. In primary human peripheral blood lymphocytes (PBL), expression of A3G and REDD1 was found to be stimulated by phytohemagglutinin (PHA) and interleukin-2. Small interfering RNA (siRNA)-mediated depletion of A3G in PHA-stimulated PBL reduced REDD1 expression and increased viral titers, which corroborates our findings in Vero cells. Silencing of REDD1 also increased viral titers, confirming the antiviral role of REDD1. Finally, pharmacological inhibition of mTORC1 by rapamycin in PHA-stimulated PBL reduced viral replication to the level found in unstimulated lymphocytes, indicating that mTORC1 activity supports MV replication as a proviral host factor. Knowledge about host factors supporting or restricting virus replication is required for a deeper understanding of virus-cell interactions and may eventually provide the basis for therapeutic intervention. This work was undertaken predominantly to explain the mechanism of A3G-mediated inhibition of MV, a negative-strand RNA virus that is not affected by the deaminase activity of A3G acting on single-stranded DNA. We found that A3G regulates the expression of several cellular proteins, which influences the capacity of the host cell to replicate MV. One of these, REDD1, which modulates the cellular metabolism in a central position by regulating the kinase complex mTORC1, was identified as the major cellular factor impairing MV replication. These findings show interesting aspects of the function of A3G and the dependence of the MV replication on the metabolic state of the cell. Interestingly, pharmacological inhibition of mTORC1 can be utilized to inhibit MV replication in Vero cells and primary human peripheral blood lymphocytes.

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Clearance of Measles Virus from Persistently Infected Cells by Short Hairpin RNA

Cited by 12 publications

References 41 publications

Morbillivirus Experimental Animal Models: Measles Virus Pathogenesis Insights from Canine Distemper Virus

Morbillivirus Experimental Animal Models: Measles Virus Pathogenesis Insights from Canine Distemper Virus

Measles virus infection of the CNS: human disease, animal models, and approaches to therapy

APOBEC3G-Regulated Host Factors Interfere with Measles Virus Replication: Role of REDD1 and Mammalian TORC1 Inhibition

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