Abstract
Multiple TonB dependent transporters (TBDTs) contribute to bacterial virulence due to the importance roles that their substrates play in bacterial growth, and possess vaccine potential. A putative TBDT, YncD, had been identified as one of in vivo induced antigens during human infection of typhoid fever, and is required for the pathogenicity of Salmonella enterica Serovar Typhi. The present study was aimed to determine the function and immunogenicity of YncD. Homologous recombination method was used to construct an yncD-deletion mutant and cirA-iroN-fepA-deletion mutant from the wild-type S. Typhi Ty2. The growth of mutants and the wild-type strain were assessed in iron-deficient medium, as well as in human macrophage cells. Recombinant YncD protein was expressed and purified using Ni-NTA affinity chromatography and anion exchange. A mouse model was then used to evaluate the immunogenicity and protection efficacy of the recombinant YncD. Antibody levels, serum bactericidal efficiency, passive immune protection, opsonophagocysis were assayed to analyse the immunoprotection mechanism of the recombinant YncD. Our results showed that YncD is associated with the iron-uptake of S. Typhi. The yncD-deletion mutant displayed impaired growth in iron-deficient medium, comparable to that the cirA-iroN-fepA-deletion mutant did. The mutation of yncD markedly decreased bacterial growth within human macrophage cells. Moreover, subcutaneous immunization of mice with recombinant YncD elicited high levels of specific anti-YncD IgG, IgG1 and IgG2a, which protected the immunized mice against the intraperitoneal challenge of S. Typhi, and decreased bacterial burdens in the livers and spleens of the infected mice. Passive immunization using the immunized sera also efficiently protected the mice from the challenge of S. Typhi. Moreover, the immunized sera enhanced in vitro bactericidal activity of complement, and opsonophagocytosis. Our results showed that YncD displays a role in the iron-uptake of S. Typhi and possesses immunogenicity.
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References
Boulton IC, Yost MK, Anderson JE, Cornelissen CN (2000) Identification of discrete domains within gonococcal transferrin-binding protein A that are necessary for ligand binding and iron uptake functions. Infect Immun 68(12):6988–6996
Cao J, Chen D, Xu W, Chen T, Xu S, Luo J et al (2007) Enhanced protection against pneumococcal infection elicited by immunization with the combination of PspA, PspC, and ClpP. Vaccine 25(27):4996–5005
Cornelissen CN, Anderson JE, Boulton IC, Sparling PF (2000) Antigenic and sequence diversity in gonococcal transferrin-binding protein A. Infect Immun 68(8):4725–4735
de Reuse H, Vinella D, Cavazza C (2013) Common themes and unique proteins for the uptake and trafficking of nickel, a metal essential for the virulence of Helicobacter pylori. Front Cell Infect Microbiol 3:94
Goel VK, Kapil A (2001) Monoclonal antibodies against the iron regulated outer membrane proteins of Acinetobacter baumannii are bactericidal. BMC Microbiol 1:16
Hagan EC, Mobley HL (2007) Uropathogenic Escherichia coli outer membrane antigens expressed during urinary tract infection. Infect Immun 75(8):3941–3949
Hennigar SR, McClung JP (2016) Nutritional immunity: starving pathogens of Trace minerals. Am J Lifestyle Med 10(3):170–173
Hu Y, Cong Y, Li S, Rao X, Wang G, Hu F (2009) Identification of in vivo induced protein antigens of Salmonella enterica Serovar Typhi during human infection. Sci China C Life Sci 52(10):942–948
Kingsley RA, Reissbrodt R, Rabsch W, Ketley JM, Tsolis RM, Everest P et al (1999) Ferrioxamine-mediated Iron(III) utilization by Salmonella enterica. Appl Environ Microbiol 65(4):1610–1618
Krewulak KD, Vogel HJ (2011) TonB or not TonB: is that the question? Biochem Cell Biol 89(2):87–97
Kumar P, Sannigrahi S, Tzeng YL (2012) The Neisseria meningitidis ZnuD zinc receptor contributes to interactions with epithelial cells and supports heme utilization when expressed in Escherichia coli. Infect Immun 80(2):657–667
Leduc I, Banks KE, Fortney KR, Patterson KB, Billings SD, Katz BP et al (2008) Evaluation of the repertoire of the TonB-dependent receptors of Haemophilus ducreyi for their role in virulence in humans. J Infect Dis 197(8):1103–1109
Lu F, Miao S, Tu J, Ni X, Xing L, Yu H et al (2013) The role of TonB-dependent receptor TbdR1 in Riemerella anatipestifer in iron acquisition and virulence. Vet Microbiol 167(3–4):713–718
Masri HP, Cornelissen CN (2002) Specific ligand binding attributable to individual epitopes of gonococcal transferrin binding protein A. Infect Immun 70(2):732–740
Mastropasqua MC, D’Orazio M, Cerasi M, Pacello F, Gismondi A, Canini A et al (2017) Growth of Pseudomonas aeruginosa in zinc poor environments is promoted by a nicotianamine-related metallophore. Mol Microbiol 106(4):543–561
Mellata M, Mitchell NM, Schodel F, Curtiss RR, Pier GB (2016) Novel vaccine antigen combinations elicit protective immune responses against Escherichia coli sepsis. Vaccine 34(5):656–662
Mobley HL, Alteri CJ (2015) Development of a vaccine against Escherichia coli urinary tract infections. Pathogens. ;5(1)
Ni Z, Chen Y, Ong E, He Y (2017) Antibiotic resistance determinant-focused Acinetobacter baumannii vaccine designed using reverse vaccinology. Int J Mol Sci. ;18(2)
Nielubowicz GR, Smith SN, Mobley HL (2008) Outer membrane antigens of the uropathogen Proteus mirabilis recognized by the humoral response during experimental murine urinary tract infection. Infect Immun 76(9):4222–4231
Nikaido H (2003) Molecular basis of bacterial outer membrane permeability revisited. Microbiol Mol Biol Rev 67(4):593–656
Noinaj N, Guillier M, Barnard TJ, Buchanan SK (2010) TonB-dependent transporters: regulation, structure, and function. Annu Rev Microbiol 64:43–60
Rabsch W, Methner U, Voigt W, Tschape H, Reissbrodt R, Williams PH (2003) Role of receptor proteins for enterobactin and 2,3-dihydroxybenzoylserine in virulence of Salmonella enterica. Infect Immun 71(12):6953–6961
Schalk IJ, Mislin GL, Brillet K (2012) Structure, function and binding selectivity and stereoselectivity of siderophore-iron outer membrane transporters. Curr Top Membr 69:37–66
Schauer K, Rodionov DA, de Reuse H (2008) New substrates for TonB-dependent transport: do we only see the ‘tip of the iceberg’? Trends Biochem Sci 33(7):330–338
Sen B, Meeker A, Ramakrishnan G (2010) The fslE homolog, FTL_0439 (fupA/B), mediates siderophore-dependent iron uptake in Francisella tularensis LVS. Infect Immun 78(10):4276–4285
Sen-Kilic E, Blackwood CB, Boehm DT, Witt WT, Malkowski AC, Bevere JR et al (2019) Intranasal peptide-based FpvA-KLH conjugate vaccine protects mice from Pseudomonas aeruginosa Acute Murine Pneumonia. Front Immunol 10:2497
Solis N, Cordwell SJ (2011) Current methodologies for proteomics of bacterial surface-exposed and cell envelope proteins. Proteomics 11(15):3169–3189
Stork M, Bos MP, Jongerius I, de Kok N, Schilders I, Weynants VE et al (2010) An outer membrane receptor of Neisseria meningitidis involved in zinc acquisition with vaccine potential. PLoS Pathog 6:e1000969
Stork M, Grijpstra J, Bos MP, Manas Torres C, Devos N, Poolman JT et al (2013) Zinc piracy as a mechanism of Neisseria meningitidis for evasion of nutritional immunity. PLoS Pathog 9(10):e1003733
Sun YY, Sun L (2015) Pseudomonas fluorescens: iron-responsive proteins and their involvement in host infection. Vet Microbiol 176(3–4):309–320
Wandersman C, Delepelaire P (2004) Bacterial iron sources: from siderophores to hemophores. Annu Rev Microbiol 58:611–647
Wang JY, Pasetti MF, Noriega FR, Anderson RJ, Wasserman SS, Galen JE et al (2001) Construction, genotypic and phenotypic characterization, and immunogenicity of attenuated DeltaguaBA Salmonella enterica Serovar Typhi strain CVD 915. Infect Immun 69(8):4734–4741
Wang J, Xiong K, Pan Q, He W, Cong Y (2020) Application of TonB-Dependent transporters in Vaccine Development of Gram-negative Bacteria. Front Cell Infect Microbiol 10:589115
Wellawa DH, Allan B, White AP, Koster W (2020) Iron-Uptake systems of Chicken-Associated Salmonella Serovars and their role in colonizing the avian host. Microorganisms. ;8(8)
Wieser A, Romann E, Magistro G, Hoffmann C, Norenberg D, Weinert K et al (2010) A multiepitope subunit vaccine conveys protection against extraintestinal pathogenic Escherichia coli in mice. Infect Immun 78(8):3432–3442
Williams PH, Rabsch W, Methner U, Voigt W, Tschape H, Reissbrodt R (2006) Catecholate receptor proteins in Salmonella enterica: role in virulence and implications for vaccine development. Vaccine 24(18):3840–3844
Xiong K, Chen Z, Xiang G, Wang J, Rao X, Hu F et al (2012) Deletion of yncD gene in Salmonella enterica ssp. Enterica Serovar Typhi leads to attenuation in mouse model. FEMS Microbiol Lett 328(1):70–77
Xiong K, Chen Z, Zhu C, Li J, Hu X, Rao X et al (2015) Safety and immunogenicity of an attenuated Salmonella enterica serovar Paratyphi A vaccine candidate. Int J Med Microbiol 305(6):563–571
Xiong K, Zhu C, Chen Z, Zheng C, Tan Y, Rao X et al (2017) Vi capsular polysaccharide produced by recombinant Salmonella enterica Serovar Paratyphi A confers immunoprotection against infection by Salmonella enterica Serovar Typhi. Front Cell Infect Microbiol 7:135
Zeng L, Wang D, Hu N, Zhu Q, Chen K, Dong K et al (2017) A Novel Pan-genome Reverse Vaccinology Approach employing a negative-selection strategy for Screening Surface-exposed antigens against leptospirosis. Front Microbiol 8:396
Acknowledgements
This work was supported by the National Natural Science Foundation of China (Grant number 31500116, 81974299), GuangDong Basic and Applied Basic Research Foundation (Grant number 2023A1515010938) and Dongguan Science and Technology Commissioner Project (20231800500552). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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Conceptualization, X.R. and Y.C.; methodology, K.X., L.D., X.R., and Y.C.; validation, L.D., Z.L., H.G., J.C., M.H.; investigation, L.D., Z.L., H.G., J.C., M.H., and K.X.; data curation, X.R. and Y.C.; writing—original draft preparation, K.X. and Y.C.; writing—review and editing, X.R., H.G., and Y.C.; supervision, Y.C.; project administration, X.R. and Y.C.; funding acquisition, K.X. and Y.C. All authors have read and agreed to the published version of the manuscript.
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Xiong, K., Deng, L., Li, Z. et al. A TonB dependent transporter YncD of Salmonella enterica Serovar Typhi possesses vaccine potential. World J Microbiol Biotechnol 40, 131 (2024). https://doi.org/10.1007/s11274-024-03937-9
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DOI: https://doi.org/10.1007/s11274-024-03937-9