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Title: Outer membrane vesicles displaying engineered glycotopes elicit protective antibodies

Abstract

The O-antigen polysaccharide (O-PS) component of lipopolysaccharides on the surface of gram-negative bacteria is both a virulence factor and a B-cell antigen. Antibodies elicited by O-PS often confer protection against infection; therefore, O-PS glycoconjugate vaccines have proven useful against a number of different pathogenic bacteria. However, conventional methods for natural extraction or chemical synthesis of O-PS are technically demanding, inefficient, and expensive. Here, we describe an alternative methodology for producing glycoconjugate vaccines whereby recombinant O-PS biosynthesis is coordinated with vesiculation in laboratory strains of Escherichia coli to yield glycosylated outer membrane vesicles (glycOMVs) decorated with pathogen-mimetic glycotopes. Using this approach, glycOMVs corresponding to eight different pathogenic bacteria were generated. For example, expression of a 17-kb O-PS gene cluster from the highly virulent Francisella tularensis subsp. tularensis (type A) strain Schu S4 in hypervesiculating E. coli cells yielded glycOMVs that displayed F. tularensis O-PS. Immunization of BALB/c mice with glycOMVs elicited significant titers of O-PS–specific serum IgG antibodies as well as vaginal and bronchoalveolar IgA antibodies. Importantly, glycOMVs significantly prolonged survival upon subsequent challenge with F. tularensis Schu S4 and provided complete protection against challenge with two different F. tularensis subsp. holarctica (type B) live vaccine strains, thereby demonstrating the vaccinemore » potential of glycOMVs. Given the ease with which recombinant glycotopes can be expressed on OMVs, the strategy described here could be readily adapted for developing vaccines against many other bacterial pathogens.« less

Authors:
ORCiD logo; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Univ. of Georgia, Athens, GA (United States); Cornell Univ., Ithaca, NY (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22); National Science Foundation (NSF); National Inst. of Health (NIH) (United States); New York State Office of Science, Technology and Academic Research (NYSTAR) (United States); US Army Research Office (ARO); Midwest Regional Center of Excellence (MRCE) for Biodefense and Emerging Infectious Disease Research (United States)
Contributing Org.:
Univ. of Iowa, Iowa City, IA (United States); Univ. of Texas, Austin, TX (United States)
OSTI Identifier:
1256136
Alternate Identifier(s):
OSTI ID: 1349046
Grant/Contract Number:  
FG02-93ER20097; DMR-1120296; CBET 1159581; CBET 1264701; GM088905-01; EB005669-01; AI044642; AI057160; GM008629; AI064184; AI076322; GM10349010; W911NF-12-1-0390
Resource Type:
Published Article
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Volume: 113 Journal Issue: 26; Journal ID: ISSN 0027-8424
Publisher:
Proceedings of the National Academy of Sciences
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; O-antigen polysaccharide; anti-glycan antibodies; glycan; glycoconjugate vaccine; humoral immune response

Citation Formats

Chen, Linxiao, Valentine, Jenny L., Huang, Chung-Jr, Endicott, Christine E., Moeller, Tyler D., Rasmussen, Jed A., Fletcher, Joshua R., Boll, Joseph M., Rosenthal, Joseph A., Dobruchowska, Justyna, Wang, Zhirui, Heiss, Christian, Azadi, Parastoo, Putnam, David, Trent, M. Stephen, Jones, Bradley D., and DeLisa, Matthew P. Outer membrane vesicles displaying engineered glycotopes elicit protective antibodies. United States: N. p., 2016. Web. doi:10.1073/pnas.1518311113.
Chen, Linxiao, Valentine, Jenny L., Huang, Chung-Jr, Endicott, Christine E., Moeller, Tyler D., Rasmussen, Jed A., Fletcher, Joshua R., Boll, Joseph M., Rosenthal, Joseph A., Dobruchowska, Justyna, Wang, Zhirui, Heiss, Christian, Azadi, Parastoo, Putnam, David, Trent, M. Stephen, Jones, Bradley D., & DeLisa, Matthew P. Outer membrane vesicles displaying engineered glycotopes elicit protective antibodies. United States. doi:10.1073/pnas.1518311113.
Chen, Linxiao, Valentine, Jenny L., Huang, Chung-Jr, Endicott, Christine E., Moeller, Tyler D., Rasmussen, Jed A., Fletcher, Joshua R., Boll, Joseph M., Rosenthal, Joseph A., Dobruchowska, Justyna, Wang, Zhirui, Heiss, Christian, Azadi, Parastoo, Putnam, David, Trent, M. Stephen, Jones, Bradley D., and DeLisa, Matthew P. Mon . "Outer membrane vesicles displaying engineered glycotopes elicit protective antibodies". United States. doi:10.1073/pnas.1518311113.
@article{osti_1256136,
title = {Outer membrane vesicles displaying engineered glycotopes elicit protective antibodies},
author = {Chen, Linxiao and Valentine, Jenny L. and Huang, Chung-Jr and Endicott, Christine E. and Moeller, Tyler D. and Rasmussen, Jed A. and Fletcher, Joshua R. and Boll, Joseph M. and Rosenthal, Joseph A. and Dobruchowska, Justyna and Wang, Zhirui and Heiss, Christian and Azadi, Parastoo and Putnam, David and Trent, M. Stephen and Jones, Bradley D. and DeLisa, Matthew P.},
abstractNote = {The O-antigen polysaccharide (O-PS) component of lipopolysaccharides on the surface of gram-negative bacteria is both a virulence factor and a B-cell antigen. Antibodies elicited by O-PS often confer protection against infection; therefore, O-PS glycoconjugate vaccines have proven useful against a number of different pathogenic bacteria. However, conventional methods for natural extraction or chemical synthesis of O-PS are technically demanding, inefficient, and expensive. Here, we describe an alternative methodology for producing glycoconjugate vaccines whereby recombinant O-PS biosynthesis is coordinated with vesiculation in laboratory strains of Escherichia coli to yield glycosylated outer membrane vesicles (glycOMVs) decorated with pathogen-mimetic glycotopes. Using this approach, glycOMVs corresponding to eight different pathogenic bacteria were generated. For example, expression of a 17-kb O-PS gene cluster from the highly virulent Francisella tularensis subsp. tularensis (type A) strain Schu S4 in hypervesiculating E. coli cells yielded glycOMVs that displayed F. tularensis O-PS. Immunization of BALB/c mice with glycOMVs elicited significant titers of O-PS–specific serum IgG antibodies as well as vaginal and bronchoalveolar IgA antibodies. Importantly, glycOMVs significantly prolonged survival upon subsequent challenge with F. tularensis Schu S4 and provided complete protection against challenge with two different F. tularensis subsp. holarctica (type B) live vaccine strains, thereby demonstrating the vaccine potential of glycOMVs. Given the ease with which recombinant glycotopes can be expressed on OMVs, the strategy described here could be readily adapted for developing vaccines against many other bacterial pathogens.},
doi = {10.1073/pnas.1518311113},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 26,
volume = 113,
place = {United States},
year = {2016},
month = {6}
}

Journal Article:
Free Publicly Available Full Text
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DOI: 10.1073/pnas.1518311113

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Cited by: 11 works
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Works referenced in this record:

T Cell-Independent Antigens Type 2
journal, April 1995


Exploitation of bacterial N -linked glycosylation to develop a novel recombinant glycoconjugate vaccine against Francisella tularensis
journal, May 2013

  • Cuccui, Jon; Thomas, Rebecca M.; Moule, Madeleine G.
  • Open Biology, Vol. 3, Issue 5
  • DOI: 10.1098/rsob.130002

Biosynthesis and Assembly of Capsular Polysaccharides in Escherichia coli
journal, June 2006


γδ T Cells Provide an Early Source of Interferon γ in Tumor Immunity
journal, August 2003

  • Gao, Yunfei; Yang, Wancai; Pan, Meng
  • The Journal of Experimental Medicine, Vol. 198, Issue 3
  • DOI: 10.1084/jem.20030584

Working toward the Future: Insights into Francisella tularensis Pathogenesis and Vaccine Development
journal, November 2009

  • Pechous, R. D.; McCarthy, T. R.; Zahrt, T. C.
  • Microbiology and Molecular Biology Reviews, Vol. 73, Issue 4
  • DOI: 10.1128/MMBR.00028-09

Conjugate vaccines
journal, October 2003


Discovery of new biosynthetic pathways: the lipid A story
journal, October 2008

  • Raetz, Christian R. H.; Guan, Ziqiang; Ingram, Brian O.
  • Journal of Lipid Research, Vol. 50, Issue Supplement
  • DOI: 10.1194/jlr.R800060-JLR200

Incorporation of Heterologous Outer Membrane and Periplasmic Proteins into Escherichia coli Outer Membrane Vesicles
journal, October 2003

  • Kesty, Nicole C.; Kuehn, Meta J.
  • Journal of Biological Chemistry, Vol. 279, Issue 3
  • DOI: 10.1074/jbc.M307628200

Biological Functions and Biogenesis of Secreted Bacterial Outer Membrane Vesicles
journal, October 2010


Recent developments in bacterial protein glycan coupling technology and glycoconjugate vaccine design
journal, April 2012

  • Terra, V. S.; Mills, D. C.; Yates, L. E.
  • Journal of Medical Microbiology, Vol. 61, Issue Pt_7
  • DOI: 10.1099/jmm.0.039438-0

Lipopolysaccharide Endotoxins
journal, June 2002


Role of antibody to lipopolysaccharide in protection against low- and high-virulence strains of Francisella tularensis
journal, August 2001


Microbial biosynthesis of designer outer membrane vesicles
journal, October 2014


Expression cloning of Yersinia enterocolitica 0 : 3 rfb gene cluster in Escherichia coli K12
journal, January 1991


Carbohydrate vaccines: developing sweet solutions to sticky situations?
journal, April 2010

  • Astronomo, Rena D.; Burton, Dennis R.
  • Nature Reviews Drug Discovery, Vol. 9, Issue 4
  • DOI: 10.1038/nrd3012

Release of the type I secreted alpha-haemolysin via outer membrane vesicles from Escherichia coli
journal, January 2006


Vaccines based on the cell surface carbohydrates of pathogenic bacteria
journal, June 2005


T-cell–B-cell cooperation
journal, April 2004

  • Mitchison, N. A.
  • Nature Reviews Immunology, Vol. 4, Issue 4
  • DOI: 10.1038/nri1334

Cognate Stimulatory B-Cell–T-Cell Interactions Are Critical for T-Cell Help Recruited by Glycoconjugate Vaccines
journal, December 1999


Escherichia coli tol-pal Mutants Form Outer Membrane Vesicles
journal, September 1998


Immunology of bacterial polysaccharide antigens
journal, November 2003


Adjuvant properties of meningococcal outer membrane vesicles and the use of adjuvants in Neisseria meningitidis protein vaccines
journal, March 2011

  • Sanders, Holly; Feavers, Ian M.
  • Expert Review of Vaccines, Vol. 10, Issue 3
  • DOI: 10.1586/erv.11.10

Recombinant outer membrane vesicles carrying Chlamydia muridarum HtrA induce antibodies that neutralize chlamydial infection in vitro
journal, January 2013

  • Bartolini, Erika; Ianni, Elvira; Frigimelica, Elisabetta
  • Journal of Extracellular Vesicles, Vol. 2, Issue 1
  • DOI: 10.3402/jev.v2i0.20181

Delivery of foreign antigens by engineered outer membrane vesicle vaccines
journal, January 2010

  • Chen, D. J.; Osterrieder, N.; Metzger, S. M.
  • Proceedings of the National Academy of Sciences, Vol. 107, Issue 7
  • DOI: 10.1073/pnas.0805532107

Structure of the O-antigen of Francisella tularensis strain 15
journal, July 1991

  • Vinogradov, Eugeny V.; Shashkov, Alexander S.; Knirel, Yuriy A.
  • Carbohydrate Research, Vol. 214, Issue 2
  • DOI: 10.1016/0008-6215(91)80036-M

Modulating the innate immune response by combinatorial engineering of endotoxin
journal, January 2013

  • Needham, Brittany D.; Carroll, Sean M.; Giles, David K.
  • Proceedings of the National Academy of Sciences, Vol. 110, Issue 4
  • DOI: 10.1073/pnas.1218080110

Identification, Characterization and Immunogenicity of an O-Antigen Capsular Polysaccharide of Francisella tularensis
journal, July 2010


B Cell Mitogenic Properties of Thymus-independent Antigens
journal, September 1973


Tularaemia: bioterrorism defence renews interest in Francisella tularensis
journal, December 2004

  • Oyston, Petra C. F.; Sjöstedt, Anders; Titball, Richard W.
  • Nature Reviews Microbiology, Vol. 2, Issue 12
  • DOI: 10.1038/nrmicro1045

Production of glycoprotein vaccines in Escherichia coli
journal, January 2010

  • Ihssen, Julian; Kowarik, Michael; Dilettoso, Sandro
  • Microbial Cell Factories, Vol. 9, Issue 1
  • DOI: 10.1186/1475-2859-9-61

Production of Secretory and Extracellular N-Linked Glycoproteins in Escherichia coli
journal, December 2010

  • Fisher, Adam C.; Haitjema, Charles H.; Guarino, Cassandra
  • Applied and Environmental Microbiology, Vol. 77, Issue 3
  • DOI: 10.1128/AEM.01901-10

Characterization of bacteria using its O-antigen with surface-enhanced Raman scattering
journal, January 2010

  • Osorio-Román, I. O.; Aroca, R. F.; Astudillo, J.
  • The Analyst, Vol. 135, Issue 8
  • DOI: 10.1039/c0an00061b

Pathogenicity and molecular genetics of O-specific side-chain lipopolysaccharides of Escherichia coli
journal, July 1992

  • Valvano, Miguel A.
  • Canadian Journal of Microbiology, Vol. 38, Issue 7
  • DOI: 10.1139/m92-117

Immunologic Memory Induced by a Glycoconjugate Vaccine in a Murine Adoptive Lymphocyte Transfer Model
journal, May 1998


How Bacterial Carbohydrates Influence the Adaptive Immune System
journal, March 2010


Somatic antigens of Shigella: Structure of the O-specific polysaccharide chain of the Shigella dysenteriae type 7 lipopolysaccharide
journal, August 1988


A Novel Role for Plasmin-Mediated Degradation of Opsonizing Antibody in the Evasion of Host Immunity by Virulent, but Not Attenuated, Francisella tularensis
journal, September 2009

  • Crane, Deborah D.; Warner, Shayna L.; Bosio, Catharine M.
  • The Journal of Immunology, Vol. 183, Issue 7
  • DOI: 10.4049/jimmunol.0901655

Engineering N-linked protein glycosylation with diverse O antigen lipopolysaccharide structures in Escherichia coli
journal, February 2005

  • Feldman, M. F.; Wacker, M.; Hernandez, M.
  • Proceedings of the National Academy of Sciences, Vol. 102, Issue 8
  • DOI: 10.1073/pnas.0500044102

Innate B cell helpers reveal novel types of antibody responses
journal, January 2013

  • Vinuesa, Carola G.; Chang, Pheh-Ping
  • Nature Immunology, Vol. 14, Issue 2
  • DOI: 10.1038/ni.2511

A systematic nomenclature for carbohydrate fragmentations in FAB-MS/MS spectra of glycoconjugates
journal, January 1988

  • Domon, Bruno; Costello, Catherine E.
  • Glycoconjugate Journal, Vol. 5, Issue 4
  • DOI: 10.1007/BF01049915

Evaluation of the establishment of herd immunity in the population by means of serological surveys and vaccination coverage
journal, February 2012

  • Plans-Rubió, Pedro
  • Human Vaccines & Immunotherapeutics, Vol. 8, Issue 2
  • DOI: 10.4161/hv.18444

Putting endotoxin to work for us: Monophosphoryl lipid A as a safe and effective vaccine adjuvant
journal, August 2008


Substrate specificity of bacterial oligosaccharyltransferase suggests a common transfer mechanism for the bacterial and eukaryotic systems
journal, April 2006

  • Wacker, M.; Feldman, M. F.; Callewaert, N.
  • Proceedings of the National Academy of Sciences, Vol. 103, Issue 18
  • DOI: 10.1073/pnas.0509207103

Engineered Bacterial Outer Membrane Vesicles with Enhanced Functionality
journal, June 2008

  • Kim, Jae-Young; Doody, Anne M.; Chen, David J.
  • Journal of Molecular Biology, Vol. 380, Issue 1
  • DOI: 10.1016/j.jmb.2008.03.076

Successful Protection against Tularemia in C57BL/6 Mice Is Correlated with Expansion of Francisella tularensis-Specific Effector T Cells
journal, November 2014

  • Griffin, Amanda J.; Crane, Deborah D.; Wehrly, Tara D.
  • Clinical and Vaccine Immunology, Vol. 22, Issue 1
  • DOI: 10.1128/CVI.00648-14

New vaccine platform?
journal, March 2010

  • Flemming, Alexandra
  • Nature Reviews Drug Discovery, Vol. 9, Issue 3
  • DOI: 10.1038/nrd3117

Francisella tularensis Schu S4 Lipopolysaccharide Core Sugar and O-Antigen Mutants Are Attenuated in a Mouse Model of Tularemia
journal, January 2014

  • Rasmussen, Jed A.; Post, Deborah M. B.; Gibson, Bradford W.
  • Infection and Immunity, Vol. 82, Issue 4
  • DOI: 10.1128/IAI.01640-13

Expression of Heterologous Antigens in Commensal Neisseria spp.: Preservation of Conformational Epitopes with Vaccine Potential
journal, October 2004