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. In this paper, 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 demonstratingmore » the vaccine potential of glycOMVs. Finally, 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:

Chen, Linxiao  ^[1];

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Valentine, Jenny L. ^[1]; Huang, Chung-Jr ^[1]; Endicott, Christine E. ^[1]; Moeller, Tyler D. ^[1]; Rasmussen, Jed A. ^[2]; Fletcher, Joshua R. ^[3]; Boll, Joseph M. ^[4]; Rosenthal, Joseph A. ^[5]; Dobruchowska, Justyna ^[6]; Wang, Zhirui ^[6]; Heiss, Christian ^[6]; Azadi, Parastoo ^[6]; Putnam, David ^[7]; Trent, M. Stephen ^[4]; Jones, Bradley D. ^[8]; DeLisa, Matthew P. ^[7]

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+ Show Author Affiliations

1. Robert Frederick Smith School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY 14853,
2. Department of Microbiology, University of Iowa, Iowa City, IA 52242,
3. Genetics Program, University of Iowa, Iowa City, IA 52242,
4. Department of Molecular Biosciences, The University of Texas at Austin, Austin, TX 78712,, Department of Infectious Diseases, The University of Georgia College of Veterinary Medicine, Athens, GA 30602,
5. Nancy E. and Peter C. Meinig School of Biomedical Engineering, Cornell University, Ithaca, NY 14853,
6. Complex Carbohydrate Research Center, The University of Georgia, Athens, GA 30602
7. Robert Frederick Smith School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY 14853,, Nancy E. and Peter C. Meinig School of Biomedical Engineering, Cornell University, Ithaca, NY 14853,
8. Department of Microbiology, University of Iowa, Iowa City, IA 52242,, Genetics Program, University of Iowa, Iowa City, IA 52242,

Publication Date:

2016-06-06

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); 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