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      Immunogenicity and protective efficacy against Treponema pallidum in New Zealand rabbits immunized with plasmid DNA encoding flagellin

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          Abstract

          Plasmid DNA encoding flagellin FlaB3 was used as a vaccination candidate for the evaluation of immunogenicity and protection against Treponema pallidum subsp. pallidum dissemination. First, intramuscular injection of the flagellin encoded by the plasmid DNA into New Zealand rabbits elicited both humoral and cellular immune responses. Total IgG production increased in response to flagellin. In addition, serum IFN-γ secretion and CD8+ cells were substantially greater in the rabbits immunized with the plasmid encoding flagellin FlaB3 than those in the rabbits immunized with recombinant flagellin. The flagellin encoded by the plasmid DNA induced significant upregulation of serum IL-6 and IL-8 compared to that of the control rabbits. Subsequently, intradermal challenge of the vaccinated New Zealand rabbits with 1 × 10 7 T. pallidum resulted in a significant reduction of the bacterial organ burden in the blood, liver, spleen, and testicles in the flagellin plasmid DNA-vaccinated rabbits. Furthermore, the histopathological analysis demonstrated that the rabbits immunized with the plasmid DNA-encoded flagellin (FlaB3) showed better immune protection. These findings provide evidence that plasmid DNA-encoded flagellin (FlaB3) may be useful as a potential immunization route for future development of a vaccine to inhibit T. pallidum dissemination in related animals.

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          Most cited references45

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          Biological basis for syphilis.

          Syphilis is a chronic sexually transmitted disease caused by Treponema pallidum subsp. pallidum. Clinical manifestations separate the disease into stages; late stages of disease are now uncommon compared to the preantibiotic era. T. pallidum has an unusually small genome and lacks genes that encode many metabolic functions and classical virulence factors. The organism is extremely sensitive to environmental conditions and has not been continuously cultivated in vitro. Nonetheless, T. pallidum is highly infectious and survives for decades in the untreated host. Early syphilis lesions result from the host's immune response to the treponemes. Bacterial clearance and resolution of early lesions results from a delayed hypersensitivity response, although some organisms escape to cause persistent infection. One factor contributing to T. pallidum's chronicity is the paucity of integral outer membrane proteins, rendering intact organisms virtually invisible to the immune system. Antigenic variation of TprK, a putative surface-exposed protein, is likely to contribute to immune evasion. T. pallidum remains exquisitely sensitive to penicillin, but macrolide resistance has recently been identified in a number of geographic regions. The development of a syphilis vaccine, thus far elusive, would have a significant positive impact on global health.
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            Evasion of Toll-like receptor 5 by flagellated bacteria.

            Toll-like receptor 5 (TLR5) recognizes an evolutionarily conserved site on bacterial flagellin that is required for flagellar filament assembly and motility. The alpha and epsilon Proteobacteria, including the important human pathogens Campylobacter jejuni, Helicobacter pylori, and Bartonella bacilliformis, require flagellar motility to efficiently infect mammalian hosts. In this study, we demonstrate that these bacteria make flagellin molecules that are not recognized by TLR5. We map the site responsible for TLR5 evasion to amino acids 89-96 of the N-terminal D1 domain, which is centrally positioned within the previously defined TLR5 recognition site. Salmonella flagellin is strongly recognized by TLR5, but mutating residues 89-96 to the corresponding H. pylori flaA sequence abolishes TLR5 recognition and also destroys bacterial motility. To preserve bacterial motility, alpha and epsilon Proteobacteria possess compensatory amino acid changes in other regions of the flagellin molecule, and we engineer a mutant form of Salmonella flagellin that evades TLR5 but retains motility. These results suggest that TLR5 evasion is critical for the survival of this subset of bacteria at mucosal sites in animals and raise the intriguing possibility that flagellin receptors provided the selective force to drive the evolution of these unique subclasses of bacterial flagellins.
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              Bacterial flagellins: mediators of pathogenicity and host immune responses in mucosa.

              Flagella contribute to the virulence of pathogenic bacteria through chemotaxis, adhesion to and invasion of host surfaces. Flagellin is the structural protein that forms the major portion of flagellar filaments. Thus, flagellin consists of a conserved domain that is widespread in bacterial species and is dedicated to filament polymerization. Conversely, mammalian hosts detect the conserved domain on flagellin monomers through Toll-like receptor (TLR) 5, which triggers proinflammatory and adaptive immune responses. This review describes the relationships among flagellin molecular structure, bacterial virulence and host defenses, with special emphasis on mucosal tissues.
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                Author and article information

                Contributors
                +86-13707340050 , yimouwu@sina.com
                Journal
                Emerg Microbes Infect
                Emerg Microbes Infect
                Emerging Microbes & Infections
                Nature Publishing Group UK (London )
                2222-1751
                7 November 2018
                7 November 2018
                2018
                : 7
                : 177
                Affiliations
                [1 ]ISNI 0000 0001 0266 8918, GRID grid.412017.1, Institute of Pathogenic Biology, Medical College, , University of South China, Hunan Province Cooperative Innovation Center for Molecular Target New Drug Study, ; Hengyang, 421001 China
                [2 ]ISNI 0000 0004 1798 5993, GRID grid.413432.3, Clinical Laboratory, , The Second Affiliated Hospital of University of South China, ; Hengyang, 421001 China
                [3 ]ISNI 0000 0001 0266 8918, GRID grid.412017.1, Department of Experimental Zoology, Medical College, , University of South China, ; Hengyang, 421001 China
                Article
                176
                10.1038/s41426-018-0176-0
                6220273
                30405111
                65a82c2c-e2d2-4964-a07f-8a2215f0dd9b
                © The Author(s) 2018

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 23 April 2018
                : 10 September 2018
                : 13 September 2018
                Funding
                Funded by: FundRef https://doi.org/10.13039/501100001809, National Natural Science Foundation of China (National Science Foundation of China);
                Award ID: 81471576, 81702046, 81701577, and 81373230
                Award ID: 81471576, 81702046, 81701577, and 81373230
                Award ID: 81471576, 81702046, 81701577, and 81373230
                Award ID: 81471576, 81702046, 81701577, and 81373230
                Award ID: 81471576, 81702046, 81701577, and 81373230
                Award ID: 81471576, 81702046, 81701577, and 81373230
                Award ID: 81471576, 81702046, 81701577, and 81373230
                Award ID: 81471576, 81702046, 81701577, and 81373230
                Award ID: 81471576, 81702046, 81701577, and 81373230
                Award ID: 81471576, 81702046, 81701577, and 81373230
                Award ID: 81471576, 81702046, 81701577, and 81373230
                Award Recipient :
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                © The Author(s) 2018

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