Respiratory Syncytial Virus Fusion Protein-Induced Toll-Like Receptor 4 (TLR4) Signaling Is Inhibited by the TLR4 Antagonists <named-content content-type="genus-species">Rhodobacter sphaeroides</named-content> Lipopolysaccharide and Eritoran (E5564) and Requires Direct Interaction with MD-2

ABSTRACT Respiratory syncytial virus (RSV) is a leading cause of infant mortality worldwide. Toll-like receptor 4 (TLR4), a signaling receptor for structurally diverse microbe-associated molecular patterns, is activated by the RSV fusion (F) protein and by bacterial lipopolysaccharide (LPS) in a CD1...

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Autores principales:	Prasad Rallabhandi, Rachel L. Phillips, Marina S. Boukhvalova, Lioubov M. Pletneva, Kari Ann Shirey, Theresa L. Gioannini, Jerrold P. Weiss, Jesse C. Chow, Lynn D. Hawkins, Stefanie N. Vogel, Jorge C. G. Blanco
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2012
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/79ea6ff9ae2848cdaa97e9031ac10480
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spelling	oai:doaj.org-article:79ea6ff9ae2848cdaa97e9031ac104802021-11-15T15:39:09ZRespiratory Syncytial Virus Fusion Protein-Induced Toll-Like Receptor 4 (TLR4) Signaling Is Inhibited by the TLR4 Antagonists <named-content content-type="genus-species">Rhodobacter sphaeroides</named-content> Lipopolysaccharide and Eritoran (E5564) and Requires Direct Interaction with MD-210.1128/mBio.00218-122150-7511https://doaj.org/article/79ea6ff9ae2848cdaa97e9031ac104802012-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00218-12https://doaj.org/toc/2150-7511ABSTRACT Respiratory syncytial virus (RSV) is a leading cause of infant mortality worldwide. Toll-like receptor 4 (TLR4), a signaling receptor for structurally diverse microbe-associated molecular patterns, is activated by the RSV fusion (F) protein and by bacterial lipopolysaccharide (LPS) in a CD14-dependent manner. TLR4 signaling by LPS also requires the presence of an additional protein, MD-2. Thus, it is possible that F protein-mediated TLR4 activation relies on MD-2 as well, although this hypothesis has not been formally tested. LPS-free RSV F protein was found to activate NF-κB in HEK293T transfectants that express wild-type (WT) TLR4 and CD14, but only when MD-2 was coexpressed. These findings were confirmed by measuring F-protein-induced interleukin 1β (IL-1β) mRNA in WT versus MD-2−/− macrophages, where MD-2−/− macrophages failed to show IL-1β expression upon F-protein treatment, in contrast to the WT. Both Rhodobacter sphaeroides LPS and synthetic E5564 (eritoran), LPS antagonists that inhibit TLR4 signaling by binding a hydrophobic pocket in MD-2, significantly reduced RSV F-protein-mediated TLR4 activity in HEK293T-TLR4–CD14–MD-2 transfectants in a dose-dependent manner, while TLR4-independent NF-κB activation by tumor necrosis factor alpha (TNF-α) was unaffected. In vitro coimmunoprecipitation studies confirmed a physical interaction between native RSV F protein and MD-2. Further, we demonstrated that the N-terminal domain of the F1 segment of RSV F protein interacts with MD-2. These data provide new insights into the importance of MD-2 in RSV F-protein-mediated TLR4 activation. Thus, targeting the interaction between MD-2 and RSV F protein may potentially lead to novel therapeutic approaches to help control RSV-induced inflammation and pathology. IMPORTANCE This study shows for the first time that the fusion (F) protein of respiratory syncytial virus (RSV), a major cause of bronchiolitis and death, particularly in infants and young children, physically interacts with the Toll-like receptor 4 (TLR4) coreceptor, MD-2, through its N-terminal domain. We show that F protein-induced TLR4 activation can be blocked by lipid A analog antagonists. This observation provides a strong experimental rationale for testing such antagonists in animal models of RSV infection for potential use in people.Prasad RallabhandiRachel L. PhillipsMarina S. BoukhvalovaLioubov M. PletnevaKari Ann ShireyTheresa L. GioanniniJerrold P. WeissJesse C. ChowLynn D. HawkinsStefanie N. VogelJorge C. G. BlancoAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 3, Iss 4 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Prasad Rallabhandi Rachel L. Phillips Marina S. Boukhvalova Lioubov M. Pletneva Kari Ann Shirey Theresa L. Gioannini Jerrold P. Weiss Jesse C. Chow Lynn D. Hawkins Stefanie N. Vogel Jorge C. G. Blanco Respiratory Syncytial Virus Fusion Protein-Induced Toll-Like Receptor 4 (TLR4) Signaling Is Inhibited by the TLR4 Antagonists <named-content content-type="genus-species">Rhodobacter sphaeroides</named-content> Lipopolysaccharide and Eritoran (E5564) and Requires Direct Interaction with MD-2
description	ABSTRACT Respiratory syncytial virus (RSV) is a leading cause of infant mortality worldwide. Toll-like receptor 4 (TLR4), a signaling receptor for structurally diverse microbe-associated molecular patterns, is activated by the RSV fusion (F) protein and by bacterial lipopolysaccharide (LPS) in a CD14-dependent manner. TLR4 signaling by LPS also requires the presence of an additional protein, MD-2. Thus, it is possible that F protein-mediated TLR4 activation relies on MD-2 as well, although this hypothesis has not been formally tested. LPS-free RSV F protein was found to activate NF-κB in HEK293T transfectants that express wild-type (WT) TLR4 and CD14, but only when MD-2 was coexpressed. These findings were confirmed by measuring F-protein-induced interleukin 1β (IL-1β) mRNA in WT versus MD-2−/− macrophages, where MD-2−/− macrophages failed to show IL-1β expression upon F-protein treatment, in contrast to the WT. Both Rhodobacter sphaeroides LPS and synthetic E5564 (eritoran), LPS antagonists that inhibit TLR4 signaling by binding a hydrophobic pocket in MD-2, significantly reduced RSV F-protein-mediated TLR4 activity in HEK293T-TLR4–CD14–MD-2 transfectants in a dose-dependent manner, while TLR4-independent NF-κB activation by tumor necrosis factor alpha (TNF-α) was unaffected. In vitro coimmunoprecipitation studies confirmed a physical interaction between native RSV F protein and MD-2. Further, we demonstrated that the N-terminal domain of the F1 segment of RSV F protein interacts with MD-2. These data provide new insights into the importance of MD-2 in RSV F-protein-mediated TLR4 activation. Thus, targeting the interaction between MD-2 and RSV F protein may potentially lead to novel therapeutic approaches to help control RSV-induced inflammation and pathology. IMPORTANCE This study shows for the first time that the fusion (F) protein of respiratory syncytial virus (RSV), a major cause of bronchiolitis and death, particularly in infants and young children, physically interacts with the Toll-like receptor 4 (TLR4) coreceptor, MD-2, through its N-terminal domain. We show that F protein-induced TLR4 activation can be blocked by lipid A analog antagonists. This observation provides a strong experimental rationale for testing such antagonists in animal models of RSV infection for potential use in people.
format	article
author	Prasad Rallabhandi Rachel L. Phillips Marina S. Boukhvalova Lioubov M. Pletneva Kari Ann Shirey Theresa L. Gioannini Jerrold P. Weiss Jesse C. Chow Lynn D. Hawkins Stefanie N. Vogel Jorge C. G. Blanco
author_facet	Prasad Rallabhandi Rachel L. Phillips Marina S. Boukhvalova Lioubov M. Pletneva Kari Ann Shirey Theresa L. Gioannini Jerrold P. Weiss Jesse C. Chow Lynn D. Hawkins Stefanie N. Vogel Jorge C. G. Blanco
author_sort	Prasad Rallabhandi
title	Respiratory Syncytial Virus Fusion Protein-Induced Toll-Like Receptor 4 (TLR4) Signaling Is Inhibited by the TLR4 Antagonists <named-content content-type="genus-species">Rhodobacter sphaeroides</named-content> Lipopolysaccharide and Eritoran (E5564) and Requires Direct Interaction with MD-2
title_short	Respiratory Syncytial Virus Fusion Protein-Induced Toll-Like Receptor 4 (TLR4) Signaling Is Inhibited by the TLR4 Antagonists <named-content content-type="genus-species">Rhodobacter sphaeroides</named-content> Lipopolysaccharide and Eritoran (E5564) and Requires Direct Interaction with MD-2
title_full	Respiratory Syncytial Virus Fusion Protein-Induced Toll-Like Receptor 4 (TLR4) Signaling Is Inhibited by the TLR4 Antagonists <named-content content-type="genus-species">Rhodobacter sphaeroides</named-content> Lipopolysaccharide and Eritoran (E5564) and Requires Direct Interaction with MD-2
title_fullStr	Respiratory Syncytial Virus Fusion Protein-Induced Toll-Like Receptor 4 (TLR4) Signaling Is Inhibited by the TLR4 Antagonists <named-content content-type="genus-species">Rhodobacter sphaeroides</named-content> Lipopolysaccharide and Eritoran (E5564) and Requires Direct Interaction with MD-2
title_full_unstemmed	Respiratory Syncytial Virus Fusion Protein-Induced Toll-Like Receptor 4 (TLR4) Signaling Is Inhibited by the TLR4 Antagonists <named-content content-type="genus-species">Rhodobacter sphaeroides</named-content> Lipopolysaccharide and Eritoran (E5564) and Requires Direct Interaction with MD-2
title_sort	respiratory syncytial virus fusion protein-induced toll-like receptor 4 (tlr4) signaling is inhibited by the tlr4 antagonists <named-content content-type="genus-species">rhodobacter sphaeroides</named-content> lipopolysaccharide and eritoran (e5564) and requires direct interaction with md-2
publisher	American Society for Microbiology
publishDate	2012
url	https://doaj.org/article/79ea6ff9ae2848cdaa97e9031ac10480
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Respiratory Syncytial Virus Fusion Protein-Induced Toll-Like Receptor 4 (TLR4) Signaling Is Inhibited by the TLR4 Antagonists <named-content content-type="genus-species">Rhodobacter sphaeroides</named-content> Lipopolysaccharide and Eritoran (E5564) and Requires Direct Interaction with MD-2

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