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PLOS ONE  2013 

Shigella flexneri 3a Outer Membrane Protein C Epitope Is Recognized by Human Umbilical Cord Sera and Associated with Protective Activity

DOI: 10.1371/journal.pone.0070539

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Abstract:

Shigella flexneri 3a is one of the five major strains of the Shigella genus responsible for dysentery, especially among children, in regions of high poverty and poor sanitation. The outer membrane proteins (OMP) of this bacterium elicit immunological responses and are considered a prime target for vaccine development. When injected into mice they elicit a protective immunological response against a lethal dose of the pathogen. The OMPs from S. flexneri 3a were isolated and resolved by two-dimension-SDS-PAGE. Two 38-kDa spots were of particular interest since in our earlier studies OMPs of such molecular mass were found to interact with umbilical cord sera. These two spots were identified as OmpC by ESI-MS/MS spectrometry. By DNA sequencing, the ompC gene from S. flexneri 3a was identical to ompC from S. flexneri 2a [Gene Bank: 24113600]. A 3D model of OmpC was built and used to predict B-cell type (discontinuous) antigenic epitopes. Six epitopes bearing the highest score were selected and the corresponding peptides were synthesized. Only the peptides representing loop V of OmpC reacted strongly with the umbilical cord serum immunoglobulins. To determine which amino acids are essential for the antigenic activity of the epitope, the loop V was scanned with a series of dodecapeptides. The peptide RYDERY was identified as a minimal sequence for the loop V epitope. Truncation at either the C- or N-terminus rendered this peptide inactive. Apart from C-terminal tyrosine, substitution of each of the remaining five amino acids with glycine, led to a precipitous loss of immunological activity. This peptide may serve as a ligand in affinity chromatography of OmpC-specific antibodies and as a component of a vaccine designed to boost human immune defenses against enterobacterial infections.

References

[1]  Black RE, Cousens S, Johnson HL, Lawn JE, Rudan I, et al. (2010) Global, regional, and national causes of child mortality in 2008: a systematic analysis. Lancet 375: 1969–1987.
[2]  Kotloff KL, Winickoff JP, Ivanoff B, Clemens JD, Swerdlow DL, et al. (1997) Global burden of Shigella infections: implications for vaccine development and implementation of control strategies. Bull World Health Organ 77: 651–66.
[3]  Peng X, Luo W, Zhang J, Wang S, Lin S (2002) Rapid detection of Shigella species in environmental sewage by an immunocapture PCR with universal primers. Appl Environ Microbiol 68: 2580–2583.
[4]  Paterson DL (2006) Resistance in gram-negative bacteria: Enterobacteriaceae. Am J Med 119: S20–S28.
[5]  Anders K, Li A, Zhao ZC, Karlsson K, Minh NB, et al. (1995) Safety and immunogenicity of the auxotrophic Shigella flexneri 2a vaccine SFL1070 with a deleted aroD gene in adult Swedish volunteers. Vaccine 13: 88–89.
[6]  Coster TS, Hoge CW, VanDeVerg LL, Hartman AB, Oaks EV, et al. (1999) Vaccination against shigellosis with attenuated Shigella flexneri 2a strain SC602. Infect Immun 67: 3437–3443.
[7]  Katz DE, Coster TS, Wolf MK, Trespalacios FC, Cohen D, et al. (2004) Two studies evaluating the safety and immunogenicity of a live, attenuated Shigella flexneri 2a vaccine (SC602) and excretion of vaccine organisms in North American volunteers. Infect Immun 72: 923–930.
[8]  Ranallo RT, Fonseka CP, Cassels F, Srinivasan J, Venkatesan MM (2005) Construction and characterization of bivalent Shigella flexneri 2a vaccine strains SC608(pCFAI) and SC608(pCFAI/LTB) that express antigens from enterotoxigenic Escherichia coli. Infect Immun 73: 258–267.
[9]  Mukhopadhaya A, Mahalanabis D, Khanam J, Chakrabarti MK (2003) Protective efficacy of oral immunization with heat-killed Shigella flexneri 2a in animal model: study of cross protection, immune response and antigenic recognition. Vaccine 21: 3043–3050.
[10]  Mukhopadhaya A, Mahalanabis D, Chakrabarti MK (2006) Role of Shigella flexneri 2a 34 kDa outer membrane protein in induction of protective immune response. Vaccine 24: 6028–6036.
[11]  Jennison AV, Verma NK (2004) Shigella flexneri infection: pathogenesis and vaccine development. FEMS Microbiol Rev 28: 43–58.
[12]  Kotloff KL, Simon JK, Pasetti MF, Sztein MB, Wooden SL, et al. (2007) Safety and Immunogenicity of CVD 1208S, a Live, Oral _guaBA _sen _set Shigella flexneri 2a Vaccine Grown on Animal-Free Media. Human Vaccine 3: 268–275.
[13]  Simon JK, Maciel M Jr, Weld ED, Wahid R, Pasetti MF, et al. (2011) Antigen-specific IgA B memory cell responses to Shigella antigens elicited in volunteers immunized with live attenuated Shigella flexneri 2a oral vaccine candidates. Clin Immunol 139: 185–192.
[14]  Biswas T (2000) Role of porin of Shigella dysenteriae type 1 in modulation of lipopolysaccharide mediated nitric oxide and interleukin-1 release by murine peritoneal macrophages. FEMS Immunol. Med. Microbiol. 29: 129–136.
[15]  Sansonetti PJ (2001) Microbes and microbial toxins: paradigms for microbial mucosal interactions III. Shigellosis: from symptoms to molecular pathogenesis. Am J Physiol Gastrointest Liver Physiol 280: G319–G323.
[16]  Koebnik R, Locher KP, Van Gelder P (2000) Structure and function of bacterial outer membrane proteins: barrels and nutshell. Mol Microbiol 37: 239–253.
[17]  Lin J, Huang S, Zhang Q (2002) Outer membrane proteins: key players for bacterial adaptation in host niches. Microbes Infect 4: 325–331.
[18]  Adamus G, Mulczyk M, Witkowska D, Romanowska E (1980) Protection against keratoconjunctivitis shigellosa induced by immunization with outer membrane proteins of Shigella spp. Infect Immun 30: 321–324.
[19]  Mulczyk M, Adamus G, Witkowska D, Romanowska E (1981) Studies on virulence of Shigella flexneri. Protective effect of outer membrane proteins. Arch Immunol Ther Exp 29: 85–90.
[20]  Witkowska D, Czarny A, Mulczyk M (1986) Humoral response in mice immunized with outer membrane proteins of Shigella flexneri. Arch Immunol Ther Exp 34: 499–504.
[21]  Witkowska D, Czarny A, Mulczyk M (1992) Humoral response in mice immunized with outer membrane proteins of Hafnia alvei. Protective activity of anti-OMP-antibodies. Arch Immunol Ther Exp 40: 301–304.
[22]  Witkowska D, Mleczko J, Mulczyk M (1985) Transfer of immunity by means of spleen cells from mice immunized with outer membrane proteins of Shigella flexneri. Arch Immunol Ther Exp 33: 629–635.
[23]  Witkowska D, Mas?owska E, Staniszewska M, Szostko B, Jankowski A, et al. (2006) Enterobacterial 38-kDa outer membrane protein is an age-dependent molecular marker of innate immunity and immunoglobulin deficiency as results from its reactivity with IgG and IgA antibody. FEMS Immunol Med Microbiol 48: 205–214.
[24]  Roy S, Das AB, Ghosh AN, Biswas T (1994) Purification, pore-forming ability, and antigenic relatedness of the major outer membrane protein of Shigella dysenteriae type 1. Infect Immun 62: 4333–4338.
[25]  Pore D, Mahata N, Pal A, Chakrabarti MK (2011) Outer membrane protein A (OmpA) of Shigella flexneri 2a, induces protective immune response in a mouse model. PLoS ONE 6(7): e22663.
[26]  Ying T, Wang H, Li M, Wang J, Wang J, et al. (2005) Immunoproteomics of outer membrane proteins and extracellular proteins of Shigella flexneri 2a 2457T. Proteomics 5: 4777–4793.
[27]  Benz I, Schmidt MA (2002) Never say never again: protein glycosylation in pathogenic bacteria. Mol Microbiol. 45: 267–276.
[28]  Baslé A, Rummel G, Storici P, Rosenbusch JP, Schirmer T (2006) Crystal structure of osmoporin OmpC from E. coli at 2.0 A. J Mol Biol. 362: 933–942.
[29]  Arnold K, Bordoli L, Kopp J, Schwede T (2006) The SWISS-MODEL Workspace: A web-based environment for protein structure homology modelling. Bioinformatics 22: 195–201.
[30]  Schwede T, Kopp J, Guex N, Peitsch MC (2003) SWISS-MODEL: An automated protein homology-modeling server. Nucleic Acids Res 31: 3381–3385.
[31]  Guex N, Peitsch MC (1997) SWISS-MODEL and the Swiss-PdbViewer: an environment for comparative protein modeling. Electrophoresis 18: 2714–2723.
[32]  Sweredoski M, Baldi P (2008) PEPITO: improved discontinuous B-cell epitope prediction using multiple distance thresholds and half sphere exposure. Bioinformatics. 24: 1459–1460 http://pepito.proteomics.ics.uci.edu.
[33]  Jarz?b A, Witkowska D, Szostko B, Hirnle L, Gamian A (2011) Potential carrier for conjugate vaccines: OMP-38 protein from Shigella flexneri 3a – isolation and purification strategies. Sepsis 4: 39–45.
[34]  Fujita Y, Taguchi H (2011) Current status of multiple antigen-presenting peptide vaccine systems: Application of organic and inorganic nanoparticles. Chem Cent J 23 5(1): 48.
[35]  Wieser A, Romann E, Magistro G, Hoffmann C, N?renberg D, et al. (2010) A multiepitope subunit vaccine conveys protection against extraintestinal pathogenic Escherichia coli in mice. Infect Immun 78: 3432–3442.
[36]  Williams KM, Bigley EC 3rd, Raybourne RB (2000) Identification of murine B-cell and T-cell epitopes of Escherichia coli outer membrane protein F with synthetic polypeptides. Infect Immun 68: 2535–2545.
[37]  Borrelli S, Hossany RB, Pinto BM (2008) Immunological evidence for functional rather than structural mimicry by a Shigella flexneri Y polysaccharide-mimetic peptide. Clin Vaccine Immunol 15: 1106–1114.
[38]  Sesardic D (1993) Synthetic peptide vaccines. J Med Microbiol 39: 241–242.
[39]  Nardin E (2010) The past decade in malaria synthetic peptide vaccine clinical trials. Hum Vaccin 6: 27–38.
[40]  McCarty M (1973) Bacterial and mycotic infections in Davis B. D., Dulbecco R., Eisen H. N., Ginsberg H. S., Wood W. B. Jr: Microbiology, Harper and Row Publishers, Hagerstown, (M)., N. York-Evanston-San Francisco.
[41]  Baldridge JR, Crane RT (1999) Monophosphoryl Lipid A (MPL) Formulations for the Next Generation of Vaccines. Methods 19: 103–107.
[42]  Arcidiacono S, Butler MM, Mello CM (2002) A rapid selective extraction procedure for the outer membrane protein (OmpF) from Escherichia coli. Protein Expr Purif 25: 134–137.
[43]  Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193: 265–275.
[44]  Laemmli UK (1970) Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 277: 680–685.
[45]  Shevchenko A, Wilm M, Vorm O, Mann M (1996) Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. Anal Chem 68: 850–858.
[46]  Gharahdaghi F, Weinberg CR, Meagher DA, Imai BS, Mische SM (1999) Mass spectrometric identification of proteins from silver-stained polyacrylamide gel: a method for the removal of silver ions to enhance sensitivity. Electrophoresis. 20: 601–605.
[47]  Carter JM (1994) Epitope mapping of a protein using the Geysen (PEPSCAN) procedure. Methods Mol Biol 36: 207–223.

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