Further Characterization of the Capsule-Like Complex (CLC) Produced by Francisella tularensis Subspecies tularensis: Protective Efficacy and Similarity to Outer Membrane Vesicles

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dc.contributor.authorChampion, Anna E.en
dc.contributor.authorBandara, Aloka B.en
dc.contributor.authorMohapatra, Nrusinghen
dc.contributor.authorFulton, Kelly M.en
dc.contributor.authorTwine, Susan M.en
dc.contributor.authorInzana, Thomas J.en
dc.date.accessioned2018-07-24T17:01:37Zen
dc.date.available2018-07-24T17:01:37Zen
dc.date.issued2018-06-15en
dc.description.abstractFrancisella tularensis is the etiologic agent of tularemia, and subspecies tularensis (type A) is the most virulent subspecies. The live vaccine strain (LVS) of subspecies holarctica produces a capsule-like complex (CLC) that consists of a large variety of glycoproteins. Expression of the CLC is greatly enhanced when the bacteria are subcultured in and grown on chemically defined medium. Deletion of two genes responsible for CLC glycosylation in LVS results in an attenuated mutant that is protective against respiratory tularemia in a mouse model. We sought to further characterize the CLC composition and to determine if a type A CLC glycosylationmutant would be attenuated inmice. The CLCs isolated from LVS extracted with 0.5% phenol or 1M urea were similar, as determined by gel electrophoresis and Western blotting, but the CLC extracted with urea was more water-soluble. The CLC extracted with either 0.5% phenol or 1M urea from type A strains was also similar to the CLC of LVS in antigenic properties, electrophoretic profile, and by transmission electron microscopy (TEM). The solubility of the CLC could be further enhanced by fractionation with Triton X-114 followed by N-Lauroylsarcosine detergents; the largest (>250 kDa) molecular size component appeared to be an aggregate of smaller components. Outer membrane vesicles/tubules (OMV/T) isolated by differential centrifugation and micro-filtration appeared similar to the CLC by TEM, and many of the proteins present in the OMV/T were also identified in soluble and insoluble fractions of the CLC. Further investigation is warranted to assess the relationship between OMV/T and the CLC. The CLC conjugated to keyhole limpet hemocyanin or flagellin was highly protective against high-dose LVS intradermal challenge and partially protective against intranasal challenge. A protective response was associated with a significant rise in cytokines IL-12, IL-10, and IFN-γ. However, a type A CLC glycosylation mutant remained virulent in BALB/cmice, and immunization with the CLC did not protectmice against high dose respiratory challenge with type A strain SCHU S4.en
dc.description.sponsorshipNational Institute of Allergy and Infection Diseasesen
dc.description.sponsorshipNIAID: U54-AI57168en
dc.description.sponsorshipVirginia-Maryland College of Veterinary Medicineen
dc.description.sponsorshipTyler J. and Frances F. Young endowmenten
dc.description.sponsorshipCenter for Plant and Microbial Complex Carbohydratesen
dc.description.sponsorshipCCRC: DFFG02-93ER-20097en
dc.format.mimetypeapplication/pdfen
dc.identifier.doihttps://doi.org/10.3389/fcimb.2018.00182en
dc.identifier.urihttp://hdl.handle.net/10919/84311en
dc.identifier.volume8en
dc.language.isoenen
dc.publisherFrontiersen
dc.rightsCreative Commons Attribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectFrancisella tularensisen
dc.subjectvirulence factorsen
dc.subjectcapsule-like complexen
dc.subjectouter membrane vesiclesen
dc.subjectprotectionen
dc.subjectimmune responseen
dc.titleFurther Characterization of the Capsule-Like Complex (CLC) Produced by Francisella tularensis Subspecies tularensis: Protective Efficacy and Similarity to Outer Membrane Vesiclesen
dc.title.serialFrontiers in Cellular and Infection Microbiologyen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten

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