Inhibiting Human Parainfluenza Virus Infection by Preactivating the Cell Entry Mechanism

ABSTRACT Paramyxoviruses, specifically, the childhood pathogen human parainfluenza virus type 3, are internalized into host cells following fusion between the viral and target cell membranes. The receptor binding protein, hemagglutinin (HA)-neuraminidase (HN), and the fusion protein (F) facilitate v...

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Autores principales:	S. F. Bottom-Tanzer, K. Rybkina, J. N. Bell, C. A. Alabi, C. Mathieu, M. Lu, S. Biswas, M. Vasquez, M. Porotto, J. A. Melero, V. Más, A. Moscona
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	antiviral conformational antibody fusion activation paramyxovirus viral fusion viral glycoprotein antibody Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/673b990248d74e3c95fce6a92ebe590f
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id	oai:doaj.org-article:673b990248d74e3c95fce6a92ebe590f
record_format	dspace
institution	DOAJ
collection	DOAJ
language	EN
topic	antiviral conformational antibody fusion activation paramyxovirus viral fusion viral glycoprotein antibody Microbiology QR1-502
spellingShingle	antiviral conformational antibody fusion activation paramyxovirus viral fusion viral glycoprotein antibody Microbiology QR1-502 S. F. Bottom-Tanzer K. Rybkina J. N. Bell C. A. Alabi C. Mathieu M. Lu S. Biswas M. Vasquez M. Porotto J. A. Melero V. Más A. Moscona Inhibiting Human Parainfluenza Virus Infection by Preactivating the Cell Entry Mechanism
description	ABSTRACT Paramyxoviruses, specifically, the childhood pathogen human parainfluenza virus type 3, are internalized into host cells following fusion between the viral and target cell membranes. The receptor binding protein, hemagglutinin (HA)-neuraminidase (HN), and the fusion protein (F) facilitate viral fusion and entry into the cell through a coordinated process involving HN activation by receptor binding, which triggers conformational changes in the F protein to activate it to reach its fusion-competent state. Interfering with this process through premature activation of the F protein has been shown to be an effective antiviral strategy in vitro. Conformational changes in the F protein leading to adoption of the postfusion form of the protein—prior to receptor engagement of HN at the host cell membrane—render the virus noninfectious. We previously identified a small compound (CSC11) that implements this antiviral strategy through an interaction with HN, causing HN to activate F in an untimely process. To assess the functionality of such compounds, it is necessary to verify that the postfusion state of F has been achieved. As demonstrated by Melero and colleagues, soluble forms of the recombinant postfusion pneumovirus F proteins and of their six helix bundle (6HB) motifs can be used to generate postfusion-specific antibodies. We produced novel anti-HPIV3 F conformation-specific antibodies that can be used to assess the functionality of compounds designed to induce F activation. In this study, using systematic chemical modifications of CSC11, we synthesized a more potent derivative of this compound, CM9. Much like CSC11, CM9 causes premature triggering of the F protein through an interaction with HN prior to receptor engagement, thereby preventing fusion and subsequent infection. In addition to validating the potency of CM9 using plaque reduction, fusion inhibition, and binding avidity assays, we confirmed the transition to a postfusion conformation of F in the presence of CM9 using our novel anti-HPIV3 conformation-specific antibodies. We present both CM9 and these newly characterized postfusion antibodies as novel tools to explore and develop antiviral approaches. In turn, these advances in both our molecular toolset and our understanding of HN-F interaction will support development of more-effective antivirals. Combining the findings described here with our recently described physiologically relevant ex vivo system, we have the potential to inform the development of therapeutics to block viral infection. IMPORTANCE Paramyxoviruses, including human parainfluenza virus type 3, are internalized into host cells by fusion between viral and target cell membranes. The receptor binding protein, hemagglutinin-neuraminidase (HN), and the fusion protein (F) facilitate viral fusion and entry into cells through a process involving HN activation by receptor binding, which triggers conformational changes in F to activate it to reach its fusion-competent state. Interfering with this process through premature activation of the F protein may be an effective antiviral strategy in vitro. We identified and optimized small compounds that implement this antiviral strategy through an interaction with HN, causing HN to activate F in an untimely fashion. To address that mechanism, we produced novel anti-HPIV3 F conformation-specific antibodies that can be used to assess the functionality of compounds designed to induce F activation. Both the novel antiviral compounds that we present and these newly characterized postfusion antibodies are novel tools for the exploration and development of antiviral approaches.
format	article
author	S. F. Bottom-Tanzer K. Rybkina J. N. Bell C. A. Alabi C. Mathieu M. Lu S. Biswas M. Vasquez M. Porotto J. A. Melero V. Más A. Moscona
author_facet	S. F. Bottom-Tanzer K. Rybkina J. N. Bell C. A. Alabi C. Mathieu M. Lu S. Biswas M. Vasquez M. Porotto J. A. Melero V. Más A. Moscona
author_sort	S. F. Bottom-Tanzer
title	Inhibiting Human Parainfluenza Virus Infection by Preactivating the Cell Entry Mechanism
title_short	Inhibiting Human Parainfluenza Virus Infection by Preactivating the Cell Entry Mechanism
title_full	Inhibiting Human Parainfluenza Virus Infection by Preactivating the Cell Entry Mechanism
title_fullStr	Inhibiting Human Parainfluenza Virus Infection by Preactivating the Cell Entry Mechanism
title_full_unstemmed	Inhibiting Human Parainfluenza Virus Infection by Preactivating the Cell Entry Mechanism
title_sort	inhibiting human parainfluenza virus infection by preactivating the cell entry mechanism
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/673b990248d74e3c95fce6a92ebe590f
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spelling	oai:doaj.org-article:673b990248d74e3c95fce6a92ebe590f2021-11-15T15:55:14ZInhibiting Human Parainfluenza Virus Infection by Preactivating the Cell Entry Mechanism10.1128/mBio.02900-182150-7511https://doaj.org/article/673b990248d74e3c95fce6a92ebe590f2019-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02900-18https://doaj.org/toc/2150-7511ABSTRACT Paramyxoviruses, specifically, the childhood pathogen human parainfluenza virus type 3, are internalized into host cells following fusion between the viral and target cell membranes. The receptor binding protein, hemagglutinin (HA)-neuraminidase (HN), and the fusion protein (F) facilitate viral fusion and entry into the cell through a coordinated process involving HN activation by receptor binding, which triggers conformational changes in the F protein to activate it to reach its fusion-competent state. Interfering with this process through premature activation of the F protein has been shown to be an effective antiviral strategy in vitro. Conformational changes in the F protein leading to adoption of the postfusion form of the protein—prior to receptor engagement of HN at the host cell membrane—render the virus noninfectious. We previously identified a small compound (CSC11) that implements this antiviral strategy through an interaction with HN, causing HN to activate F in an untimely process. To assess the functionality of such compounds, it is necessary to verify that the postfusion state of F has been achieved. As demonstrated by Melero and colleagues, soluble forms of the recombinant postfusion pneumovirus F proteins and of their six helix bundle (6HB) motifs can be used to generate postfusion-specific antibodies. We produced novel anti-HPIV3 F conformation-specific antibodies that can be used to assess the functionality of compounds designed to induce F activation. In this study, using systematic chemical modifications of CSC11, we synthesized a more potent derivative of this compound, CM9. Much like CSC11, CM9 causes premature triggering of the F protein through an interaction with HN prior to receptor engagement, thereby preventing fusion and subsequent infection. In addition to validating the potency of CM9 using plaque reduction, fusion inhibition, and binding avidity assays, we confirmed the transition to a postfusion conformation of F in the presence of CM9 using our novel anti-HPIV3 conformation-specific antibodies. We present both CM9 and these newly characterized postfusion antibodies as novel tools to explore and develop antiviral approaches. In turn, these advances in both our molecular toolset and our understanding of HN-F interaction will support development of more-effective antivirals. Combining the findings described here with our recently described physiologically relevant ex vivo system, we have the potential to inform the development of therapeutics to block viral infection. IMPORTANCE Paramyxoviruses, including human parainfluenza virus type 3, are internalized into host cells by fusion between viral and target cell membranes. The receptor binding protein, hemagglutinin-neuraminidase (HN), and the fusion protein (F) facilitate viral fusion and entry into cells through a process involving HN activation by receptor binding, which triggers conformational changes in F to activate it to reach its fusion-competent state. Interfering with this process through premature activation of the F protein may be an effective antiviral strategy in vitro. We identified and optimized small compounds that implement this antiviral strategy through an interaction with HN, causing HN to activate F in an untimely fashion. To address that mechanism, we produced novel anti-HPIV3 F conformation-specific antibodies that can be used to assess the functionality of compounds designed to induce F activation. Both the novel antiviral compounds that we present and these newly characterized postfusion antibodies are novel tools for the exploration and development of antiviral approaches.S. F. Bottom-TanzerK. RybkinaJ. N. BellC. A. AlabiC. MathieuM. LuS. BiswasM. VasquezM. PorottoJ. A. MeleroV. MásA. MosconaAmerican Society for Microbiologyarticleantiviralconformational antibodyfusion activationparamyxovirusviral fusionviral glycoprotein antibodyMicrobiologyQR1-502ENmBio, Vol 10, Iss 1 (2019)

Inhibiting Human Parainfluenza Virus Infection by Preactivating the Cell Entry Mechanism

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