Characterization of Inner and Outer Membrane Proteins from Francisella tularensis Strains LVS and Schu S4 and Identification of Potential Subunit Vaccine Candidates

Deborah M B Post; Bram Slütter; Birgit Schilling; Aroon T Chande; Jed A Rasmussen; Bradley D Jones; Alexandria K D'Souza; Lorri M Reinders; John T Harty; Bradford W Gibson; Michael A Apicella

doi:10.1128/mBio.01592-17

Back

Characterization of Inner and Outer Membrane Proteins from Francisella tularensis Strains LVS and Schu S4 and Identification of Potential Subunit Vaccine Candidates

Journal article

Open access

Peer reviewed

Characterization of Inner and Outer Membrane Proteins from Francisella tularensis Strains LVS and Schu S4 and Identification of Potential Subunit Vaccine Candidates

Deborah M B Post, Bram Slütter, Birgit Schilling, Aroon T Chande, Jed A Rasmussen, Bradley D Jones, Alexandria K D'Souza, Lorri M Reinders, John T Harty, Bradford W Gibson, …

mBio, Vol.8(5), p.e01592-17

10/10/2017

DOI: 10.1128/mBio.01592-17

PMCID: PMC5635693

PMID: 29018123

Files and links (1)

url

https://doi.org/10.1128/mBio.01592-17View

Published (Version of record) Open Access

Abstract

is the causative agent of tularemia and a potential bioterrorism agent. In the present study, we isolated, identified, and quantified the proteins present in the membranes of the virulent type A strain, Schu S4, and the attenuated type B strain, LVS (live vaccine strain). Spectral counting of mass spectrometric data showed enrichment for membrane proteins in both strains. Mice vaccinated with whole LVS membranes encapsulated in poly (lactic-co-glycolic acid) (PLGA) nanoparticles containing the adjuvant polyinosinic-polycytidylic acid [poly(I·C)] showed significant protection against a challenge with LVS compared to the results seen with naive mice or mice vaccinated with either membranes or poly(I·C) alone. The PLGA-encapsulated Schu S4 membranes with poly(I·C) alone did not significantly protect mice from a lethal intraperitoneal challenge with Schu S4; however, this vaccination strategy provided protection from LVS challenge. Mice that received the encapsulated Schu S4 membranes followed by a booster of LVS bacteria showed significant protection with respect to a lethal Schu S4 challenge compared to control mice. Western blot analyses of the sera from the Schu S4-vaccinated mice that received an LVS booster showed four immunoreactive bands. One of these bands from the corresponding one-dimensional (1D) SDS-PAGE experiment represented capsule. The remaining bands were excised, digested with trypsin, and analyzed using mass spectrometry. The most abundant proteins present in these immunoreactive samples were an outer membrane OmpA-like protein, FopA; the type IV pilus fiber building block protein; a hypothetical membrane protein; and lipoproteins LpnA and Lpp3. These proteins should serve as potential targets for future recombinant protein vaccination studies. The low infectious dose, the high potential mortality/morbidity rates, and the ability to be disseminated as an aerosol make a potential agent for bioterrorism. These characteristics led the Centers for Disease Control (CDC) to classify as a Tier 1 pathogen. Currently, there is no vaccine approved for general use in the United States.

Polylactic Acid-Polyglycolic Acid Copolymer

Vaccines, Subunit - genetics

Vaccination

Tularemia - prevention & control

Francisella tularensis - chemistry

Nanoparticles

Mass Spectrometry

Tularemia - immunology

Poly I-C - immunology

Macrophages - immunology

Macrophages - microbiology

Disease Models, Animal

Francisella tularensis - immunology

Membrane Proteins - isolation & purification

Adjuvants, Immunologic

Lactic Acid

Membrane Proteins - genetics

Polyglycolic Acid

Membrane Proteins - immunology

Vaccines, Subunit - immunology

Vaccines, Attenuated - immunology

Animals

Bacterial Vaccines - immunology

Membrane Proteins - chemistry

Proteomics

Francisella tularensis - pathogenicity

Mice

Mice, Inbred BALB C

Details

Title: Subtitle: Characterization of Inner and Outer Membrane Proteins from Francisella tularensis Strains LVS and Schu S4 and Identification of Potential Subunit Vaccine Candidates
Creators: Deborah M B Post - Buck Institute for Research on Aging, Novato, California, USA dpost@buckinstitute.org michael-apicella@uiowa.edu
Bram Slütter - Department of Microbiology, the University of Iowa, Iowa City, Iowa, USA
Birgit Schilling - Buck Institute for Research on Aging, Novato, California, USA
Aroon T Chande - Department of Microbiology, the University of Iowa, Iowa City, Iowa, USA
Jed A Rasmussen - Department of Microbiology, the University of Iowa, Iowa City, Iowa, USA
Bradley D Jones - Department of Microbiology, the University of Iowa, Iowa City, Iowa, USA
Alexandria K D'Souza - Buck Institute for Research on Aging, Novato, California, USA
Lorri M Reinders - Buck Institute for Research on Aging, Novato, California, USA
John T Harty - Department of Microbiology, the University of Iowa, Iowa City, Iowa, USA
Bradford W Gibson - Department of Pharmaceutical Chemistry, University of California, San Francisco, California, USA
Michael A Apicella - Department of Microbiology, the University of Iowa, Iowa City, Iowa, USA dpost@buckinstitute.org michael-apicella@uiowa.edu
Resource Type: Journal article
Publication Details: mBio, Vol.8(5), p.e01592-17
DOI: 10.1128/mBio.01592-17
PMID: 29018123
PMCID: PMC5635693
NLM abbreviation: mBio
ISSN: 2161-2129
eISSN: 2150-7511
Publisher: United States
Grant note: S10 OD016281 / NIH HHS P01 AI044642 / NIAID NIH HHS
Language: English
Date published: 10/10/2017
Academic Unit: Microbiology and Immunology; Pathology; Internal Medicine
Record Identifier: 9984046801602771

Metrics

20 Record Views

17 Times Cited - Web of Science