Convergent Evolution of Neutralizing Antibodies to <named-content content-type="genus-species">Staphylococcus aureus</named-content> γ-Hemolysin C That Recognize an Immunodominant Primary Sequence-Dependent B-Cell Epitope

ABSTRACT Staphylococcus aureus infection is a major public health threat in part due to the spread of antibiotic resistance and repeated failures to develop a protective vaccine. Infection is associated with production of virulence factors that include exotoxins that attack host barriers and cellula...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	David N. Hernandez, Kayan Tam, Bo Shopsin, Emily E. Radke, Karen Law, Timothy Cardozo, Victor J. Torres, Gregg J. Silverman
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	B cell epitope Staphylococcus aureus antibody function bacteriophage display biotechnology epitope Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/5fd5c02cfeea46f7987a4235a59bedfc
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:5fd5c02cfeea46f7987a4235a59bedfc
record_format	dspace
spelling	oai:doaj.org-article:5fd5c02cfeea46f7987a4235a59bedfc2021-11-15T15:56:47ZConvergent Evolution of Neutralizing Antibodies to <named-content content-type="genus-species">Staphylococcus aureus</named-content> γ-Hemolysin C That Recognize an Immunodominant Primary Sequence-Dependent B-Cell Epitope10.1128/mBio.00460-202150-7511https://doaj.org/article/5fd5c02cfeea46f7987a4235a59bedfc2020-06-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00460-20https://doaj.org/toc/2150-7511ABSTRACT Staphylococcus aureus infection is a major public health threat in part due to the spread of antibiotic resistance and repeated failures to develop a protective vaccine. Infection is associated with production of virulence factors that include exotoxins that attack host barriers and cellular defenses, such as the leukocidin (Luk) family of bicomponent pore-forming toxins. To investigate the structural basis of antibody-mediated functional inactivation of Luk toxins, we generated a panel of murine monoclonal antibodies (MAbs) that neutralize host cell killing by the γ-hemolysin HlgCB. By biopanning these MAbs against a phage-display library of random Luk peptide fragments, we identified a small subregion within the rim domain of HlgC as the epitope for all the MAbs. Within the native holotoxin, this subregion folds into a conserved β-hairpin structure, with exposed key residues, His252 and Tyr253, required for antibody binding. On the basis of the phage-display results and molecular modeling, a 15-amino-acid synthetic peptide representing the minimal epitope on HlgC (HlgC241-255) was designed, and preincubation with this peptide blocked antibody-mediated HIgCB neutralization. Immunization of mice with HlgC241-255 or the homologous LukS246-260 subregion peptide elicited serum antibodies that specifically recognized the native holotoxin subunits. Furthermore, serum IgG from patients who were convalescent for invasive S. aureus infection showed neutralization of HlgCB toxin activity ex vivo, which recognized the immunodominant HlgC241-255 peptide and was dependent on His252 and Tyr253 residues. We have thus validated an efficient, rapid, and scalable experimental workflow for identification of immunodominant and immunogenic leukotoxin-neutralizing B-cell epitopes that can be exploited for new S. aureus-protective vaccines and immunotherapies.David N. HernandezKayan TamBo ShopsinEmily E. RadkeKaren LawTimothy CardozoVictor J. TorresGregg J. SilvermanAmerican Society for MicrobiologyarticleB cell epitopeStaphylococcus aureusantibody functionbacteriophage displaybiotechnologyepitopeMicrobiologyQR1-502ENmBio, Vol 11, Iss 3 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	B cell epitope Staphylococcus aureus antibody function bacteriophage display biotechnology epitope Microbiology QR1-502
spellingShingle	B cell epitope Staphylococcus aureus antibody function bacteriophage display biotechnology epitope Microbiology QR1-502 David N. Hernandez Kayan Tam Bo Shopsin Emily E. Radke Karen Law Timothy Cardozo Victor J. Torres Gregg J. Silverman Convergent Evolution of Neutralizing Antibodies to <named-content content-type="genus-species">Staphylococcus aureus</named-content> γ-Hemolysin C That Recognize an Immunodominant Primary Sequence-Dependent B-Cell Epitope
description	ABSTRACT Staphylococcus aureus infection is a major public health threat in part due to the spread of antibiotic resistance and repeated failures to develop a protective vaccine. Infection is associated with production of virulence factors that include exotoxins that attack host barriers and cellular defenses, such as the leukocidin (Luk) family of bicomponent pore-forming toxins. To investigate the structural basis of antibody-mediated functional inactivation of Luk toxins, we generated a panel of murine monoclonal antibodies (MAbs) that neutralize host cell killing by the γ-hemolysin HlgCB. By biopanning these MAbs against a phage-display library of random Luk peptide fragments, we identified a small subregion within the rim domain of HlgC as the epitope for all the MAbs. Within the native holotoxin, this subregion folds into a conserved β-hairpin structure, with exposed key residues, His252 and Tyr253, required for antibody binding. On the basis of the phage-display results and molecular modeling, a 15-amino-acid synthetic peptide representing the minimal epitope on HlgC (HlgC241-255) was designed, and preincubation with this peptide blocked antibody-mediated HIgCB neutralization. Immunization of mice with HlgC241-255 or the homologous LukS246-260 subregion peptide elicited serum antibodies that specifically recognized the native holotoxin subunits. Furthermore, serum IgG from patients who were convalescent for invasive S. aureus infection showed neutralization of HlgCB toxin activity ex vivo, which recognized the immunodominant HlgC241-255 peptide and was dependent on His252 and Tyr253 residues. We have thus validated an efficient, rapid, and scalable experimental workflow for identification of immunodominant and immunogenic leukotoxin-neutralizing B-cell epitopes that can be exploited for new S. aureus-protective vaccines and immunotherapies.
format	article
author	David N. Hernandez Kayan Tam Bo Shopsin Emily E. Radke Karen Law Timothy Cardozo Victor J. Torres Gregg J. Silverman
author_facet	David N. Hernandez Kayan Tam Bo Shopsin Emily E. Radke Karen Law Timothy Cardozo Victor J. Torres Gregg J. Silverman
author_sort	David N. Hernandez
title	Convergent Evolution of Neutralizing Antibodies to <named-content content-type="genus-species">Staphylococcus aureus</named-content> γ-Hemolysin C That Recognize an Immunodominant Primary Sequence-Dependent B-Cell Epitope
title_short	Convergent Evolution of Neutralizing Antibodies to <named-content content-type="genus-species">Staphylococcus aureus</named-content> γ-Hemolysin C That Recognize an Immunodominant Primary Sequence-Dependent B-Cell Epitope
title_full	Convergent Evolution of Neutralizing Antibodies to <named-content content-type="genus-species">Staphylococcus aureus</named-content> γ-Hemolysin C That Recognize an Immunodominant Primary Sequence-Dependent B-Cell Epitope
title_fullStr	Convergent Evolution of Neutralizing Antibodies to <named-content content-type="genus-species">Staphylococcus aureus</named-content> γ-Hemolysin C That Recognize an Immunodominant Primary Sequence-Dependent B-Cell Epitope
title_full_unstemmed	Convergent Evolution of Neutralizing Antibodies to <named-content content-type="genus-species">Staphylococcus aureus</named-content> γ-Hemolysin C That Recognize an Immunodominant Primary Sequence-Dependent B-Cell Epitope
title_sort	convergent evolution of neutralizing antibodies to <named-content content-type="genus-species">staphylococcus aureus</named-content> γ-hemolysin c that recognize an immunodominant primary sequence-dependent b-cell epitope
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/5fd5c02cfeea46f7987a4235a59bedfc
work_keys_str_mv	AT davidnhernandez convergentevolutionofneutralizingantibodiestonamedcontentcontenttypegenusspeciesstaphylococcusaureusnamedcontentghemolysincthatrecognizeanimmunodominantprimarysequencedependentbcellepitope AT kayantam convergentevolutionofneutralizingantibodiestonamedcontentcontenttypegenusspeciesstaphylococcusaureusnamedcontentghemolysincthatrecognizeanimmunodominantprimarysequencedependentbcellepitope AT boshopsin convergentevolutionofneutralizingantibodiestonamedcontentcontenttypegenusspeciesstaphylococcusaureusnamedcontentghemolysincthatrecognizeanimmunodominantprimarysequencedependentbcellepitope AT emilyeradke convergentevolutionofneutralizingantibodiestonamedcontentcontenttypegenusspeciesstaphylococcusaureusnamedcontentghemolysincthatrecognizeanimmunodominantprimarysequencedependentbcellepitope AT karenlaw convergentevolutionofneutralizingantibodiestonamedcontentcontenttypegenusspeciesstaphylococcusaureusnamedcontentghemolysincthatrecognizeanimmunodominantprimarysequencedependentbcellepitope AT timothycardozo convergentevolutionofneutralizingantibodiestonamedcontentcontenttypegenusspeciesstaphylococcusaureusnamedcontentghemolysincthatrecognizeanimmunodominantprimarysequencedependentbcellepitope AT victorjtorres convergentevolutionofneutralizingantibodiestonamedcontentcontenttypegenusspeciesstaphylococcusaureusnamedcontentghemolysincthatrecognizeanimmunodominantprimarysequencedependentbcellepitope AT greggjsilverman convergentevolutionofneutralizingantibodiestonamedcontentcontenttypegenusspeciesstaphylococcusaureusnamedcontentghemolysincthatrecognizeanimmunodominantprimarysequencedependentbcellepitope
_version_	1718427091236552704

Convergent Evolution of Neutralizing Antibodies to <named-content content-type="genus-species">Staphylococcus aureus</named-content> γ-Hemolysin C That Recognize an Immunodominant Primary Sequence-Dependent B-Cell Epitope

Ejemplares similares