Engineering Recombinant Reoviruses To Display gp41 Membrane-Proximal External-Region Epitopes from HIV-1

ABSTRACT The gp41 membrane-proximal external region (MPER) is a target for broadly neutralizing antibody responses against human immunodeficiency virus type 1 (HIV-1). However, replication-defective virus vaccines currently under evaluation in clinical trials do not efficiently elicit MPER-specific...

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Autores principales:	Karl W. Boehme, Mine' Ikizler, Jason A. Iskarpatyoti, J. Denise Wetzel, Jordan Willis, James E. Crowe, Celia C. LaBranche, David C. Montefiori, Gregory J. Wilson, Terence S. Dermody
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Lenguaje:	EN
Publicado:	American Society for Microbiology 2016
Materias:	human immunodeficiency virus immunization live vector vaccines neutralizing antibodies reovirus Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/480e9507ca4949cba0f9935e73e5b20d
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spelling	oai:doaj.org-article:480e9507ca4949cba0f9935e73e5b20d2021-11-15T15:21:17ZEngineering Recombinant Reoviruses To Display gp41 Membrane-Proximal External-Region Epitopes from HIV-110.1128/mSphere.00086-162379-5042https://doaj.org/article/480e9507ca4949cba0f9935e73e5b20d2016-06-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00086-16https://doaj.org/toc/2379-5042ABSTRACT The gp41 membrane-proximal external region (MPER) is a target for broadly neutralizing antibody responses against human immunodeficiency virus type 1 (HIV-1). However, replication-defective virus vaccines currently under evaluation in clinical trials do not efficiently elicit MPER-specific antibodies. Structural modeling suggests that the MPER forms an α-helical coiled coil that is required for function and immunogenicity. To maintain the native MPER conformation, we used reverse genetics to engineer replication-competent reovirus vectors that displayed MPER sequences in the α-helical coiled-coil tail domain of viral attachment protein σ1. Sequences in reovirus strain type 1 Lang (T1L) σ1 were exchanged with sequences encoding HIV-1 strain Ba-L MPER epitope 2F5 or the entire MPER. Individual 2F5 or MPER substitutions were introduced at virion-proximal or virion-distal sites in the σ1 tail. Recombinant reoviruses containing heterologous HIV-1 sequences were viable and produced progeny yields comparable to those with wild-type virus. HIV-1 sequences were retained following 10 serial passages in cell culture, indicating that the substitutions were genetically stable. Recombinant viruses engineered to display the 2F5 epitope or full-length MPER in σ1 were recognized by purified 2F5 antibody. Inoculation of mice with 2F5-containing vectors or rabbits with 2F5- or MPER-containing vectors elicited anti-reovirus antibodies, but HIV-1-specific antibodies were not detected. Together, these findings indicate that heterologous sequences that form α-helices can functionally replace native sequences in the α-helical tail domain of reovirus attachment protein σ1. However, although these vectors retain native antigenicity, they were not immunogenic, illustrating the difficulty of experimentally inducing immune responses to this essential region of HIV-1. IMPORTANCE Vaccines to protect against HIV-1, the causative agent of AIDS, are not approved for use. Antibodies that neutralize genetically diverse strains of HIV-1 bind to discrete regions of the envelope glycoproteins, including the gp41 MPER. We engineered recombinant reoviruses that displayed MPER epitopes in attachment protein σ1 (REO-MPER vectors). The REO-MPER vectors replicated with wild-type efficiency, were genetically stable, and retained native antigenicity. However, we did not detect HIV-1-specific immune responses following inoculation of the REO-MPER vectors into small animals. This work provides proof of principle for engineering reovirus to express antigenic epitopes and illustrates the difficulty in eliciting MPER-specific immune responses.Karl W. BoehmeMine' IkizlerJason A. IskarpatyotiJ. Denise WetzelJordan WillisJames E. CroweCelia C. LaBrancheDavid C. MontefioriGregory J. WilsonTerence S. DermodyAmerican Society for Microbiologyarticlehuman immunodeficiency virusimmunizationlive vector vaccinesneutralizing antibodiesreovirusMicrobiologyQR1-502ENmSphere, Vol 1, Iss 3 (2016)
institution	DOAJ
collection	DOAJ
language	EN
topic	human immunodeficiency virus immunization live vector vaccines neutralizing antibodies reovirus Microbiology QR1-502
spellingShingle	human immunodeficiency virus immunization live vector vaccines neutralizing antibodies reovirus Microbiology QR1-502 Karl W. Boehme Mine' Ikizler Jason A. Iskarpatyoti J. Denise Wetzel Jordan Willis James E. Crowe Celia C. LaBranche David C. Montefiori Gregory J. Wilson Terence S. Dermody Engineering Recombinant Reoviruses To Display gp41 Membrane-Proximal External-Region Epitopes from HIV-1
description	ABSTRACT The gp41 membrane-proximal external region (MPER) is a target for broadly neutralizing antibody responses against human immunodeficiency virus type 1 (HIV-1). However, replication-defective virus vaccines currently under evaluation in clinical trials do not efficiently elicit MPER-specific antibodies. Structural modeling suggests that the MPER forms an α-helical coiled coil that is required for function and immunogenicity. To maintain the native MPER conformation, we used reverse genetics to engineer replication-competent reovirus vectors that displayed MPER sequences in the α-helical coiled-coil tail domain of viral attachment protein σ1. Sequences in reovirus strain type 1 Lang (T1L) σ1 were exchanged with sequences encoding HIV-1 strain Ba-L MPER epitope 2F5 or the entire MPER. Individual 2F5 or MPER substitutions were introduced at virion-proximal or virion-distal sites in the σ1 tail. Recombinant reoviruses containing heterologous HIV-1 sequences were viable and produced progeny yields comparable to those with wild-type virus. HIV-1 sequences were retained following 10 serial passages in cell culture, indicating that the substitutions were genetically stable. Recombinant viruses engineered to display the 2F5 epitope or full-length MPER in σ1 were recognized by purified 2F5 antibody. Inoculation of mice with 2F5-containing vectors or rabbits with 2F5- or MPER-containing vectors elicited anti-reovirus antibodies, but HIV-1-specific antibodies were not detected. Together, these findings indicate that heterologous sequences that form α-helices can functionally replace native sequences in the α-helical tail domain of reovirus attachment protein σ1. However, although these vectors retain native antigenicity, they were not immunogenic, illustrating the difficulty of experimentally inducing immune responses to this essential region of HIV-1. IMPORTANCE Vaccines to protect against HIV-1, the causative agent of AIDS, are not approved for use. Antibodies that neutralize genetically diverse strains of HIV-1 bind to discrete regions of the envelope glycoproteins, including the gp41 MPER. We engineered recombinant reoviruses that displayed MPER epitopes in attachment protein σ1 (REO-MPER vectors). The REO-MPER vectors replicated with wild-type efficiency, were genetically stable, and retained native antigenicity. However, we did not detect HIV-1-specific immune responses following inoculation of the REO-MPER vectors into small animals. This work provides proof of principle for engineering reovirus to express antigenic epitopes and illustrates the difficulty in eliciting MPER-specific immune responses.
format	article
author	Karl W. Boehme Mine' Ikizler Jason A. Iskarpatyoti J. Denise Wetzel Jordan Willis James E. Crowe Celia C. LaBranche David C. Montefiori Gregory J. Wilson Terence S. Dermody
author_facet	Karl W. Boehme Mine' Ikizler Jason A. Iskarpatyoti J. Denise Wetzel Jordan Willis James E. Crowe Celia C. LaBranche David C. Montefiori Gregory J. Wilson Terence S. Dermody
author_sort	Karl W. Boehme
title	Engineering Recombinant Reoviruses To Display gp41 Membrane-Proximal External-Region Epitopes from HIV-1
title_short	Engineering Recombinant Reoviruses To Display gp41 Membrane-Proximal External-Region Epitopes from HIV-1
title_full	Engineering Recombinant Reoviruses To Display gp41 Membrane-Proximal External-Region Epitopes from HIV-1
title_fullStr	Engineering Recombinant Reoviruses To Display gp41 Membrane-Proximal External-Region Epitopes from HIV-1
title_full_unstemmed	Engineering Recombinant Reoviruses To Display gp41 Membrane-Proximal External-Region Epitopes from HIV-1
title_sort	engineering recombinant reoviruses to display gp41 membrane-proximal external-region epitopes from hiv-1
publisher	American Society for Microbiology
publishDate	2016
url	https://doaj.org/article/480e9507ca4949cba0f9935e73e5b20d
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Engineering Recombinant Reoviruses To Display gp41 Membrane-Proximal External-Region Epitopes from HIV-1

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