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. 1995 Oct;63(10):3927–3935. doi: 10.1128/iai.63.10.3927-3935.1995

Recognition of three epitopic regions on invasion plasmid antigen C by immune sera of rhesus monkeys infected with Shigella flexneri 2a.

K R Turbyfill 1, S W Joseph 1, E V Oaks 1
PMCID: PMC173552  PMID: 7558301

Abstract

The invasive ability of Shigella spp. is correlated with the expression of several plasmid-encoded proteins, including invasion plasmid antigen C (IpaC). By characterizing the antigenic structure of IpaC with monoclonal antibodies and convalescent-phase sera, it may be possible to determine the physical location of specific epitopes as well as the involvement of epitopes in a protective immune response or the host's susceptibility to disease. By using overlapping octameric synthetic peptides, which together represent the entire IpaC protein, the precise linear sequence of four surface-exposed epitopes was defined for four IpaC monoclonal antibodies. Furthermore, 17 unique peptide epitopes of IpaC were mapped by using 9-day-postinfection serum samples from 13 rhesus monkeys challenged with Shigella flexneri 2a. Each individual recognized a somewhat different array of IpaC peptide epitopes after infection with shigellae. However, the epitopes were clustered within three regions of the protein: region I (between amino acid residues 1 and 61), region II (between amino acid residues 177 and 258), and region III (between amino acid residues 298 and 307). Region II was recognized by 92% of S. flexneri-infected individuals and was considered to be a highly immunogenic region. Animals asymptomatic for shigellosis after challenge with S. flexneri recognized peptide epitopes within all three epitopic regions of IpaC, whereas symptomatic animals recognized peptides in only one or two of the epitopic regions. Antibody from monkeys challenged with S. sonnei recognized IpaC peptide epitopes which fell within and outside the three S. flexneri epitopic regions. While numerous potential epitopes exist on the IpaC protein, the identification of three regions in which epitopes are clustered suggests that these regions are significant with respect to the immune response and to subsequent pathogenesis postinfection.

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Selected References

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  1. Barzu S., Nato F., Rouyre S., Mazie J. C., Sansonetti P., Phalipon A. Characterization of B-cell epitopes on IpaB, an invasion-associated antigen of Shigella flexneri: identification of an immunodominant domain recognized during natural infection. Infect Immun. 1993 Sep;61(9):3825–3831. doi: 10.1128/iai.61.9.3825-3831.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baudry B., Maurelli A. T., Clerc P., Sadoff J. C., Sansonetti P. J. Localization of plasmid loci necessary for the entry of Shigella flexneri into HeLa cells, and characterization of one locus encoding four immunogenic polypeptides. J Gen Microbiol. 1987 Dec;133(12):3403–3413. doi: 10.1099/00221287-133-12-3403. [DOI] [PubMed] [Google Scholar]
  3. Black R. E., Levine M. M., Clements M. L., Losonsky G., Herrington D., Berman S., Formal S. B. Prevention of shigellosis by a Salmonella typhi-Shigella sonnei bivalent vaccine. J Infect Dis. 1987 Jun;155(6):1260–1265. doi: 10.1093/infdis/155.6.1260. [DOI] [PubMed] [Google Scholar]
  4. Buysse J. M., Stover C. K., Oaks E. V., Venkatesan M., Kopecko D. J. Molecular cloning of invasion plasmid antigen (ipa) genes from Shigella flexneri: analysis of ipa gene products and genetic mapping. J Bacteriol. 1987 Jun;169(6):2561–2569. doi: 10.1128/jb.169.6.2561-2569.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cohen D., Block C., Green M. S., Lowell G., Ofek I. Immunoglobulin M, A, and G antibody response to lipopolysaccharide O antigen in symptomatic and asymptomatic Shigella infections. J Clin Microbiol. 1989 Jan;27(1):162–167. doi: 10.1128/jcm.27.1.162-167.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cohen D., Green M. S., Block C., Slepon R., Ofek I. Prospective study of the association between serum antibodies to lipopolysaccharide O antigen and the attack rate of shigellosis. J Clin Microbiol. 1991 Feb;29(2):386–389. doi: 10.1128/jcm.29.2.386-389.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dinari G., Hale T. L., Austin S. W., Formal S. B. Local and systemic antibody responses to Shigella infection in rhesus monkeys. J Infect Dis. 1987 May;155(5):1065–1069. doi: 10.1093/infdis/155.5.1065. [DOI] [PubMed] [Google Scholar]
  8. Fieser T. M., Tainer J. A., Geysen H. M., Houghten R. A., Lerner R. A. Influence of protein flexibility and peptide conformation on reactivity of monoclonal anti-peptide antibodies with a protein alpha-helix. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8568–8572. doi: 10.1073/pnas.84.23.8568. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Formal S. B., Maenza R. M., Austin S., LaBrec E. H. Failure of parenteral vaccines to protect monkeys against experimental shigellosis. Proc Soc Exp Biol Med. 1967 Jun;125(2):347–349. doi: 10.3181/00379727-125-32087. [DOI] [PubMed] [Google Scholar]
  10. Formal S. B., Oaks E. V., Olsen R. E., Wingfield-Eggleston M., Snoy P. J., Cogan J. P. Effect of prior infection with virulent Shigella flexneri 2a on the resistance of monkeys to subsequent infection with Shigella sonnei. J Infect Dis. 1991 Sep;164(3):533–537. doi: 10.1093/infdis/164.3.533. [DOI] [PubMed] [Google Scholar]
  11. Getzoff E. D., Geysen H. M., Rodda S. J., Alexander H., Tainer J. A., Lerner R. A. Mechanisms of antibody binding to a protein. Science. 1987 Mar 6;235(4793):1191–1196. doi: 10.1126/science.3823879. [DOI] [PubMed] [Google Scholar]
  12. Geysen H. M., Meloen R. H., Barteling S. J. Use of peptide synthesis to probe viral antigens for epitopes to a resolution of a single amino acid. Proc Natl Acad Sci U S A. 1984 Jul;81(13):3998–4002. doi: 10.1073/pnas.81.13.3998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Geysen H. M., Rodda S. J., Mason T. J., Tribbick G., Schoofs P. G. Strategies for epitope analysis using peptide synthesis. J Immunol Methods. 1987 Sep 24;102(2):259–274. doi: 10.1016/0022-1759(87)90085-8. [DOI] [PubMed] [Google Scholar]
  14. Geysen H. M., Tainer J. A., Rodda S. J., Mason T. J., Alexander H., Getzoff E. D., Lerner R. A. Chemistry of antibody binding to a protein. Science. 1987 Mar 6;235(4793):1184–1190. doi: 10.1126/science.3823878. [DOI] [PubMed] [Google Scholar]
  15. HIGGINS A. R., FLOYD T. M., KADER M. A. Studies in shigellosis. III. A controlled evaluation of a monovalent Shigella vaccine in a highly endemic environment. Am J Trop Med Hyg. 1955 Mar;4(2):281–288. [PubMed] [Google Scholar]
  16. High N., Mounier J., Prévost M. C., Sansonetti P. J. IpaB of Shigella flexneri causes entry into epithelial cells and escape from the phagocytic vacuole. EMBO J. 1992 May;11(5):1991–1999. doi: 10.1002/j.1460-2075.1992.tb05253.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hopp T. P., Woods K. R. Prediction of protein antigenic determinants from amino acid sequences. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3824–3828. doi: 10.1073/pnas.78.6.3824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Li A., Cam P. D., Islam D., Minh N. B., Huan P. T., Rong Z. C., Karlsson K., Lindberg G., Lindberg A. A. Immune responses in Vietnamese children after a single dose of the auxotrophic, live Shigella flexneri Y vaccine strain SFL124. J Infect. 1994 Jan;28(1):11–23. doi: 10.1016/s0163-4453(94)94006-1. [DOI] [PubMed] [Google Scholar]
  19. Li A., Rong Z. C., Ekwall E., Forsum U., Lindberg A. A. Serum antibody responses against shigella lipopolysaccharides and invasion plasmid-coded antigens in shigella infected Swedish patients. Scand J Infect Dis. 1993;25(5):569–577. doi: 10.3109/00365549309008545. [DOI] [PubMed] [Google Scholar]
  20. Li A., Zhao C. R., Ekwall E., Lindberg A. A. Serum IgG antibody responses to Shigella invasion plasmid-coded antigens detected by immunoblot. Scand J Infect Dis. 1994;26(4):435–445. doi: 10.3109/00365549409008617. [DOI] [PubMed] [Google Scholar]
  21. Mills J. A., Buysse J. M., Oaks E. V. Shigella flexneri invasion plasmid antigens B and C: epitope location and characterization with monoclonal antibodies. Infect Immun. 1988 Nov;56(11):2933–2941. doi: 10.1128/iai.56.11.2933-2941.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ménard R., Sansonetti P., Parsot C., Vasselon T. Extracellular association and cytoplasmic partitioning of the IpaB and IpaC invasins of S. flexneri. Cell. 1994 Nov 4;79(3):515–525. doi: 10.1016/0092-8674(94)90260-7. [DOI] [PubMed] [Google Scholar]
  23. Oaks E. V., Hale T. L., Formal S. B. Serum immune response to Shigella protein antigens in rhesus monkeys and humans infected with Shigella spp. Infect Immun. 1986 Jul;53(1):57–63. doi: 10.1128/iai.53.1.57-63.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Oaks E. V., Turbyfill K. R. Myosin-cross-reactive epitope of Shigella flexneri invasion plasmid antigen B. Infect Immun. 1992 Feb;60(2):557–564. doi: 10.1128/iai.60.2.557-564.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Orr N., Robin G., Cohen D., Arnon R., Lowell G. H. Immunogenicity and efficacy of oral or intranasal Shigella flexneri 2a and Shigella sonnei proteosome-lipopolysaccharide vaccines in animal models. Infect Immun. 1993 Jun;61(6):2390–2395. doi: 10.1128/iai.61.6.2390-2395.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Phalipon A., Arondel J., Nato F., Rouyre S., Mazie J. C., Sansonetti P. J. Identification and characterization of B-cell epitopes of IpaC, an invasion-associated protein of Shigella flexneri. Infect Immun. 1992 May;60(5):1919–1926. doi: 10.1128/iai.60.5.1919-1926.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Phalipon A., Michetti P., Kaufmann M., Cavaillon J. M., Huerre M., Kraehenbuhl J. P., Sansonetti P. J. Protection against invasion of the mouse pulmonary epithelium by a monoclonal IgA directed against Shigella flexneri lipopolysaccharide. Ann N Y Acad Sci. 1994 Aug 15;730:356–358. doi: 10.1111/j.1749-6632.1994.tb44291.x. [DOI] [PubMed] [Google Scholar]
  28. Sankaran K., Ramachandran V., Subrahmanyam Y. V., Rajarathnam S., Elango S., Roy R. K. Congo red-mediated regulation of levels of Shigella flexneri 2a membrane proteins. Infect Immun. 1989 Aug;57(8):2364–2371. doi: 10.1128/iai.57.8.2364-2371.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Stover C. K., Marana D. P., Carter J. M., Roe B. A., Mardis E., Oaks E. V. The 56-kilodalton major protein antigen of Rickettsia tsutsugamushi: molecular cloning and sequence analysis of the sta56 gene and precise identification of a strain-specific epitope. Infect Immun. 1990 Jul;58(7):2076–2084. doi: 10.1128/iai.58.7.2076-2084.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tacket C. O., Binion S. B., Bostwick E., Losonsky G., Roy M. J., Edelman R. Efficacy of bovine milk immunoglobulin concentrate in preventing illness after Shigella flexneri challenge. Am J Trop Med Hyg. 1992 Sep;47(3):276–283. doi: 10.4269/ajtmh.1992.47.276. [DOI] [PubMed] [Google Scholar]
  31. Van de Verg L. L., Herrington D. A., Boslego J., Lindberg A. A., Levine M. M. Age-specific prevalence of serum antibodies to the invasion plasmid and lipopolysaccharide antigens of Shigella species in Chilean and North American populations. J Infect Dis. 1992 Jul;166(1):158–161. doi: 10.1093/infdis/166.1.158. [DOI] [PubMed] [Google Scholar]
  32. Venkatesan M. M., Buysse J. M., Kopecko D. J. Characterization of invasion plasmid antigen genes (ipaBCD) from Shigella flexneri. Proc Natl Acad Sci U S A. 1988 Dec;85(23):9317–9321. doi: 10.1073/pnas.85.23.9317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Venkatesan M. M., Buysse J. M., Oaks E. V. Surface presentation of Shigella flexneri invasion plasmid antigens requires the products of the spa locus. J Bacteriol. 1992 Mar;174(6):1990–2001. doi: 10.1128/jb.174.6.1990-2001.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Yao R., Palchaudhuri S. Nucleotide sequence of the ipaBCD structural genes of Shigella dysenteriae. Mol Microbiol. 1991 Sep;5(9):2217–2221. doi: 10.1111/j.1365-2958.1991.tb02151.x. [DOI] [PubMed] [Google Scholar]

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