IFN-Gamma Inhibits JC Virus Replication in Glial Cells by Suppressing T-Antigen Expression

DeSimone, Francesca I.; Sariyer, Rahsan; Otalora, Yolanda-Lopez; Yarandi, Shadan S; Craigie, Michael; Gordon, Jennifer; Sariyer, Ilker Kudret

doi:10.1371/journal.pone.0129694

Cited by 38 publications

(36 citation statements)

References 42 publications

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“…At various post-exposure time points, cells were harvested, and total RNA was extracted with an RNA extraction kit (RNeasy, Qiagen) according to the manufacturer’s instructions. RT-PCR reactions were performed as described previously (De Simone et al, 2015). After treatment with DNase I, followed by phenol/chloroform extraction and EtOH precipitation, cDNAs were synthesized using M-MuLV reverse transcriptase.…”

Section: Methodsmentioning

confidence: 99%

Alcohol-Mediated Missplicing of Mcl-1 Pre-mRNA is Involved in Neurotoxicity

Sariyer

DeSimone

Donadoni

et al. 2017

Alcohol Clin Exp Res

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Background Heavy and chronic ethanol (EtOH) exposure can cause significant structural and functional damage to the adult brain. The most devastating consequence of EtOH exposure is the neurotoxicity associated with the depletion of neurons. Regulation of splice variants in the brain can modulate protein functions, which may ultimately affect behaviors associated with alcohol dependence and EtOH-mediated neurotoxicity. Since alcohol consumption is associated with neurotoxicity, it is possible that altered splicing of survival and pro-survival factors during the development of alcoholism may contribute to the neurotoxicity. Methods Primary human neurons and a neuroblastoma cell line were exposed to different concentrations of EtOH for various time periods. Cell viability and neuronal marker expression were analyzed by MTT assay and immunoblotting, respectively. Effect of EtOH exposure on splicing regulatory protein expression and alternative splicing of candidate genes were analyzed by a biochemical approach. Transcriptional activity of SRSF1 gene was determined by reporter gene analysis. Results Our results suggest that EtOH exposure to neuronal cells at 25 mM and higher concentrations are detrimental. In addition, EtOH exposure caused a dramatic reduction in serine-arginine rich splicing factor 1 (SRSF1) expression levels. Furthermore, EtOH exposure led to pre-mRNA missplicing of Mcl-1, a pro-survival member of the Bcl-2 family, by downregulating the expression levels of serine/arginine rich splicing factor 1 (SRSF1). Moreover, ectopic expression of both SRSF1 and MCL-1L isoform was able to recover EtOH-mediated neurotoxicity. Conclusions Our results suggest that ethanol exposure can lead to pre-mRNA missplicing of Mcl-1 in neuronal cells. Our results indicate that ethanol exposure of neurons leads to a decrease in the ratio of Mcl-1L/Mcl-1S by favoring pro-apoptotic Mcl-1S splicing over anti-apoptotic Mcl-1L isoform suggesting that Mcl-1S may play a crucial role in neurotoxicity associated with alcohol consumption.

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Section: Methodsmentioning

confidence: 99%

Alcohol-Mediated Missplicing of Mcl-1 Pre-mRNA is Involved in Neurotoxicity

Sariyer

DeSimone

Donadoni

et al. 2017

Alcohol Clin Exp Res

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“…Each IFN induces a distinct ISG profile that includes both shared and unique members (56,57). These ISG proteins can exert, among numerous other activities, antiviral actions that can target vulnerabilities at all stages of the viral life cycle, including entry, transcription, translation, genome replication, assembly, and egress (reviewed in reference 58), although the described effects of most reports have concluded that the inhibition is largely at the level of viral transcription and translation, including studies on the structurally related polyomaviruses (59,60). The identified effector molecules in these studies include RNA-dependent protein kinase (PKR), adenosine deaminase (ADAR), guanylate-binding proteins (GBP1 and GBP2), and nitric oxide synthetase (NOS), although in other instances the observed effects are unascribed (61)(62)(63)(64)(65)(66)(67).…”

Section: Discussionmentioning

confidence: 99%

“…If indicated, the addition of NH 4 Cl was coincident with the addition of PsV to a final concentration of 20 mM. L2 was detected by Western blotting with the K5L2 antibody (47)(48)(49)(50)(51)(53)(54)(55)(56)(57)(58)(59)(60)(61)(62)(63)(64)(65)(66)(67).…”

Section: Methodsmentioning

confidence: 99%

“…The rabbit polyclonal antisera recognizing HPV16 and HPV45 capsids were previously described (81,82), as were the anti-L2 monoclonal antibodies K1L2 (L2 amino acids 64 to 81), used for immunofluorescent detection, and K5L2 (47)(48)(49)(50)(51)(53)(54)(55)(56)(57)(58)(59)(60)(61)(62)(63)(64)(65)(66)(67), used for Western blot detection, both kind gifts from Martin Müller (DKFZ, Heidelberg) (83). The anti-LAMP-1 monoclonal antibody (H4A3), developed by August et al, was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of NICHD and maintained by the Department of Biological Sciences at the University of Iowa (84).…”

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confidence: 99%

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Interferon Gamma Prevents Infectious Entry of Human Papillomavirus 16 via an L2-Dependent Mechanism

Day¹,

Thompson²,

Lowy³

et al. 2017

J Virol

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In this study, we report that gamma interferon (IFN-␥) treatment, but not IFN-␣, -␤, or -treatment, dramatically decreased infection of human papillomavirus 16 (HPV16) pseudovirus (PsV). In a survey of 20 additional HPV and animal papillomavirus types, we found that many, but not all, PsV types were also inhibited by IFN-␥. Microscopic and biochemical analyses of HPV16 PsV determined that the antiviral effect was exerted at the level of endosomal processing of the incoming capsid and depended on the JAK2/STAT1 pathway. In contrast to infection in the absence of IFN-␥, where L1 proteolytic products are produced during endosomal capsid processing and L2/DNA complexes segregate from L1 in the late endosome and travel to the nucleus, IFN-␥ treatment led to decreased L1 proteolysis and retention of L2 and the viral genome in the late endosome/lysosome. PsV sensitivity or resistance to IFN-␥ treatment was mapped to the L2 protein, as determined with infectious hybrid PsV, in which the L1 protein was derived from an IFN-␥-sensitive HPV type and the L2 protein from an IFN-␥-insensitive type or vice versa.IMPORTANCE A subset of HPV are the causative agents of many human cancers, most notably cervical cancer. This work describes the inhibition of infection of multiple HPV types, including oncogenic types, by treatment with IFN-␥, an antiviral cytokine that is released from stimulated immune cells. Exposure of cells to IFN-␥ has been shown to trigger the expression of proteins with broad antiviral effector functions, most of which act to prevent viral transcription or translation. Interestingly, in this study, we show that infection is blocked at the early step of virus entry into the host cell by retention of the minor capsid protein, L2, and the viral genome instead of trafficking into the nucleus. Thus, a novel antiviral mechanism for IFN-␥ has been revealed. KEYWORDS HPV, papillomavirus, interferon, IFN-␥, L2P apillomaviruses (PV) are members of a large group of nonenveloped DNA tumor viruses that infect epithelial tissues of a wide range of vertebrate species. A subset of oncogenic mucosal human PV (HPV) types are the causal agents of cervical cancer. These high-risk types, especially HPV16, are also linked to vulvar, vaginal, anal, and oropharyngeal cancers (1). The cascade of viral protein expression is integrally tied to epithelial differentiation, with virion production only occurring in the terminally differentiated upper layers (2). These features combine to allow circumvention of host innate and adaptive immune responses, as few proinflammatory signals are elicited during these early stages (3). However, most PV infections that induce overt hyperproliferation are eventually cleared with the apparent involvement of lymphocytic infiltrates (reviewed in reference 4). Also, HPV infections are often superimposed on colonization with other microbial agents, e.g., in the female reproductive tract (reviewed in reference 5). It is, therefore, of interest to determine if antiviral molecules that are known compon...

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“…Human peripheral blood mononuclear cells (PBMC) were isolated from the heparinized blood of buffy coats by Ficoll-Paque technique, induced with PMA and Ionomycin, and conditioned media from either uninduced or induced PBMCs were collected as described previously (De-Simone et al, 2015). Conditioned media were supplemented into growth media (50%) of primary human fetal glial (PHFG) cells transfected with reporter constructs or infected with JCV.…”

Section: Methodsmentioning

confidence: 99%

Immune suppression of JC virus gene expression is mediated by SRSF1

et al. 2016

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Progressive multifocal leukoemcephalopathy (PML) is a fatal demyelinating disease caused by the human neurotropic JC virus (JCV). JCV infects the majority of the human population during childhood and establishes a latent/persistent life-long infection. The virus reactivates under immunosuppressive conditions by unknown mechanisms, resulting in productive infection of oligodendrocytes in the central nervous system (CNS). Given the fact that the natural occurrence of PML is strongly associated with immunosuppression, the functional and molecular interaction between glial cells and neuroimmune signaling mediated by soluble immune mediators is likely to play a major role in reactivation of JCV and the progression of the lytic viral life cycle leading to the development of PML. In order to explore the effect of soluble immune mediators secreted by peripheral blood mononuclear cells (PBMCs) on JCV transcription, primary human fetal glial (PHFG) cells were treated with conditioned media from PBMCs. We observed a strong suppression of JCV early as well as late gene transcription in cells treated with conditioned media from induced PBMCs. Using a variety of virological and molecular biological approaches, we demonstrate that immune mediators secreted by PBMCs induce the expression of SRSF1, a strong inhibitor of JCV gene expression, and inhibit the replication of JCV. Our results show that down-regulation of SRSF1 in glial cells overcomes the suppression of JCV gene expression and its replication mediated by soluble immune mediators. These findings suggest the presence of a novel immune signaling pathway between glial cells and PBMCs that may control JCV gene expression during the course of viral reactivation.

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IFN-Gamma Inhibits JC Virus Replication in Glial Cells by Suppressing T-Antigen Expression

Cited by 38 publications

References 42 publications

Alcohol-Mediated Missplicing of Mcl-1 Pre-mRNA is Involved in Neurotoxicity

Alcohol-Mediated Missplicing of Mcl-1 Pre-mRNA is Involved in Neurotoxicity

Interferon Gamma Prevents Infectious Entry of Human Papillomavirus 16 via an L2-Dependent Mechanism

Immune suppression of JC virus gene expression is mediated by SRSF1

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