siRNA-Mediated Simultaneous Regulation of the Cellular Innate Immune Response and Human Respiratory Syncytial Virus Replication

Human respiratory syncytial virus (HRSV) infection is a common cause of severe lower respiratory tract diseases such as bronchiolitis and pneumonia. Both virus replication and the associated inflammatory immune response are believed to be behind these pathologies. So far, no vaccine or effective treatment is available for this viral infection. With the aim of finding new strategies to counteract HRSV replication and modulate the immune response, specific small interfering RNAs (siRNAs) were generated targeting the mRNA coding for the viral fusion (F) protein or nucleoprotein (N), or for two proteins involved in intracellular immune signaling, which are named tripartite motif-containing protein 25 (TRIM25) and retinoic acid-inducible gene-I (RIG-I). Furthermore, two additional bispecific siRNAs were designed that silenced F and TRIM25 (TRIM25/HRSV-F) or N and RIG-I (RIG-I/HRSV-N) simultaneously. All siRNAs targeting N or F, but not those silencing TRIM25 or RIG-I alone, significantly reduced viral titers. However, while siRNAs targeting F inhibited only the expression of the F mRNA and protein, the siRNAs targeting N led to a general inhibition of viral mRNA and protein expression. The N-targeting siRNAs also induced a drastic decrease in the expression of genes of the innate immune response. These results show that both virus replication and the early innate immune response can be regulated by targeting distinct viral products with siRNAs, which may be related to the different role of each protein in the life cycle of the virus.

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Platelets Modulate Innate Immune Response Against Human Respiratory Syncytial Virus In Vitro

Viral Immunology ◽

10.1089/vim.2016.0161 ◽

2017 ◽

Vol 30 (8) ◽

pp. 576-581 ◽

Cited By ~ 5

Author(s):

Vesla I. Kullaya ◽

Quirijn de Mast ◽

Andre van der Ven ◽

Hicham elMoussaoui ◽

Gibson Kibiki ◽

...

Keyword(s):

Immune Response ◽

Respiratory Syncytial Virus ◽

Innate Immune Response ◽

Human Respiratory Syncytial Virus ◽

Innate Immune ◽

Syncytial Virus

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Human Respiratory Syncytial Virus Nucleoprotein and Inclusion Bodies Antagonize the Innate Immune Response Mediated by MDA5 and MAVS

Journal of Virology ◽

10.1128/jvi.00215-12 ◽

2012 ◽

Vol 86 (15) ◽

pp. 8245-8258 ◽

Cited By ~ 74

Author(s):

A. W. Lifland ◽

J. Jung ◽

E. Alonas ◽

C. Zurla ◽

J. E. Crowe ◽

...

Keyword(s):

Immune Response ◽

Respiratory Syncytial Virus ◽

Innate Immune Response ◽

Inclusion Bodies ◽

Human Respiratory Syncytial Virus ◽

Innate Immune ◽

Syncytial Virus

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Human metapneumovirus induces more severe disease and stronger innate immune response in BALB/c mice as compared with respiratory syncytial virus

Respiratory Research ◽

10.1186/1465-9921-8-6 ◽

2007 ◽

Vol 8 (1) ◽

Cited By ~ 29

Author(s):

Barbara Huck ◽

Dieter Neumann-Haefelin ◽

Annette Schmitt-Graeff ◽

Markus Weckmann ◽

Jörg Mattes ◽

...

Keyword(s):

Immune Response ◽

Respiratory Syncytial Virus ◽

Innate Immune Response ◽

Severe Disease ◽

Human Metapneumovirus ◽

Innate Immune ◽

Syncytial Virus

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Bromodomain Containing Protein 4 (BRD4) Regulates Expression of its Interacting Coactivators in the Innate Response to Respiratory Syncytial Virus

Frontiers in Molecular Biosciences ◽

10.3389/fmolb.2021.728661 ◽

2021 ◽

Vol 8 ◽

Author(s):

Xiaofang Xu ◽

Morgan Mann ◽

Dianhua Qiao ◽

Yi Li ◽

Jia Zhou ◽

...

Keyword(s):

Immune Response ◽

Extracellular Matrix ◽

Transcription Factors ◽

Respiratory Syncytial Virus ◽

Innate Immune Response ◽

Airway Epithelial Cells ◽

Innate Immune ◽

Airway Epithelial ◽

Gene Regulatory ◽

Syncytial Virus

Bromodomain-containing protein 4 plays a central role in coordinating the complex epigenetic component of the innate immune response. Previous studies implicated BRD4 as a component of a chromatin-modifying complex that is dynamically recruited to a network of protective cytokines by binding activated transcription factors, polymerases, and histones to trigger their rapid expression via transcriptional elongation. Our previous study extended our understanding of the airway epithelial BRD4 interactome by identifying over 100 functionally important coactivators and transcription factors, whose association is induced by respiratory syncytial virus (RSV) infection. RSV is an etiological agent of recurrent respiratory tract infections associated with exacerbations of chronic obstructive pulmonary disease. Using a highly selective small-molecule BRD4 inhibitor (ZL0454) developed by us, we extend these findings to identify the gene regulatory network dependent on BRD4 bromodomain (BD) interactions. Human small airway epithelial cells were infected in the absence or presence of ZL0454, and gene expression profiling was performed. A highly reproducible dataset was obtained which indicated that BRD4 mediates both activation and repression of RSV-inducible gene regulatory networks controlling cytokine expression, interferon (IFN) production, and extracellular matrix remodeling. Index genes of functionally significant clusters were validated independently. We discover that BRD4 regulates the expression of its own gene during the innate immune response. Interestingly, BRD4 activates the expression of NFκB/RelA, a coactivator that binds to BRD4 in a BD-dependent manner. We extend this finding to show that BRD4 also regulates other components of its functional interactome, including the Mediator (Med) coactivator complex and the SWI/SNF-related, matrix-associated, actin-dependent regulator of chromatin (SMARC) subunits. To provide further insight into mechanisms for BRD4 in RSV expression, we mapped 7,845 RSV-inducible Tn5 transposase peaks onto the BRD4-dependent gene bodies. These were located in promoters and introns of cytostructural and extracellular matrix (ECM) formation genes. These data indicate that BRD4 mediates the dynamic response of airway epithelial cells to RNA infection by modulating the expression of its coactivators, controlling the expression of host defense mechanisms and remodeling genes through changes in promoter accessibility.

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p53-responsive TLR8 SNP enhances human innate immune response to respiratory syncytial virus

Journal of Clinical Investigation ◽

10.1172/jci128626 ◽

2019 ◽

Vol 129 (11) ◽

pp. 4875-4884 ◽

Cited By ~ 8

Author(s):

Daniel Menendez ◽

Joyce Snipe ◽

Jacqui Marzec ◽

Cynthia L. Innes ◽

Fernando P. Polack ◽

...

Keyword(s):

Immune Response ◽

Respiratory Syncytial Virus ◽

Innate Immune Response ◽

Innate Immune ◽

Syncytial Virus

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Respiratory Syncytial virus NS1 protein targets the transactivator binding domain of MED25

10.1101/2021.11.19.469356 ◽

2021 ◽

Author(s):

Vincent Basse ◽

Jiawei Dong ◽

Andressa Peres de Oliveira ◽

Pierre-Olivier Vidalain ◽

Frédéric Tangy ◽

...

Keyword(s):

Immune Response ◽

Respiratory Syncytial Virus ◽

Rna Polymerase Ii ◽

Innate Immune Response ◽

Innate Immune ◽

Transactivation Domain ◽

Ns1 Protein ◽

Syncytial Virus ◽

Acid Domain

Respiratory syncytial virus has evolved a unique strategy to evade host immune response by coding for two non-structural proteins NS1 and NS2. Recently it was shown that in infected cells, nuclear NS1 could be involved in transcription regulation of host genes linked to innate immune response, via an interaction with chromatin and the Mediator complex. Here we identified the MED25 Mediator subunit as an NS1 interactor in a yeast two-hybrid screen. We demonstrate that NS1 directly interacts with MED25 in vitro and in cellula, and that this interaction involves the C-terminal α3 helix of NS1 and the MED25 ACID domain. More specifically we showed by NMR that the NS1 α3 sequence primarily binds to the MED25 ACID H2 face, which is a transactivation domain (TAD) binding site for transcription regulators such as ATF6α, a master regulator of ER stress response activated upon viral infection. Moreover, we found out that the NS1 α3 helix could compete with ATF6α TAD binding to MED25. This finding points to a mechanism of NS1 interfering with innate immune response by impairing recruitment by cellular TADs of the Mediator via MED25 and hence transcription of specific genes by RNA polymerase II.

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Depletion of TAX1BP1 amplifies innate immune responses during respiratory syncytial virus infection

Journal of Virology ◽

10.1128/jvi.00912-21 ◽

2021 ◽

Author(s):

Delphyne Descamps ◽

Andressa Peres de Oliveira ◽

Lorène Gonnin ◽

Sarah Madrières ◽

Jenna Fix ◽

...

Keyword(s):

Immune Response ◽

Respiratory Syncytial Virus ◽

Respiratory Infections ◽

The Elderly ◽

Cellular Protein ◽

Innate Immune ◽

Efficient Treatment ◽

Rsv Infection ◽

Syncytial Virus ◽

Nucleoprotein N

Respiratory syncytial virus (RSV) is the main cause of acute respiratory infections in young children, and also has a major impact on the elderly and immunocompromised people. In the absence of a vaccine or efficient treatment, a better understanding of RSV interactions with the host antiviral response during infection is needed. Previous studies revealed that cytoplasmic inclusion bodies (IBs) where viral replication and transcription occur could play a major role in the control of innate immunity during infection by recruiting cellular proteins involved in the host antiviral response. We recently showed that the morphogenesis of IBs relies on a liquid-liquid phase separation mechanism depending on the interaction between viral nucleoprotein (N) and phosphoprotein (P). These scaffold proteins are expected to play a central role in the recruitment of cellular proteins to IBs. Here, we performed a yeast two-hybrid screen using RSV N protein as a bait, and identified the cellular protein TAX1BP1 as a potential partner of this viral protein. This interaction was validated by pulldown and immunoprecipitation assays. We showed that TAX1BP1 suppression has only a limited impact on RSV infection in cell cultures. However, RSV replication is decreased in TAX1BP1-deficient mice (TAX1BP1 KO ), whereas the production of inflammatory and antiviral cytokines is enhanced. In vitro infection of wild-type or TAX1BP1 KO alveolar macrophages confirmed that the innate immune response to RSV infection is enhanced in the absence of TAX1BP1. Altogether, our results suggest that RSV could hijack TAX1BP1 to restrain the host immune response during infection. Importance Respiratory syncytial virus (RSV), which is the leading cause of lower respiratory tract illness in infants, still remains a medical problem in the absence of vaccine or efficient treatment. This virus is also recognized as a main pathogen in the elderly and immunocompromised people, and the occurrence of co-infections (with other respiratory viruses and bacteria) amplifies the risks of developing respiratory distress. In this context, a better understanding of the pathogenesis associated to viral respiratory infections, which depends on both viral replication and the host immune response, is needed. The present study reveals that the cellular protein TAX1BP1, which interacts with the RSV nucleoprotein N, participates in the control of the innate immune response during RSV infection, suggesting that N-TAX1BP1 interaction represents a new target for the development of antivirals.

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