Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1993 Sep;61(9):3678–3687. doi: 10.1128/iai.61.9.3678-3687.1993

Synthesis, characterization, and clinical evaluation of conjugate vaccines composed of the O-specific polysaccharides of Shigella dysenteriae type 1, Shigella flexneri type 2a, and Shigella sonnei (Plesiomonas shigelloides) bound to bacterial toxoids.

D N Taylor 1, A C Trofa 1, J Sadoff 1, C Chu 1, D Bryla 1, J Shiloach 1, D Cohen 1, S Ashkenazi 1, Y Lerman 1, W Egan 1, et al.
PMCID: PMC281064  PMID: 8359890

Abstract

The theoretic basis for developing conjugate vaccines, to induce immunoglobulin G (IgG) lipopolysaccharide (LPS) antibodies for the prevention of shigellosis, has been described (J. B. Robbins, C.-Y. Chu, and R. Schneerson, Clin. Infect. Dis. 15:346-361, 1992). The O-specific polysaccharides (O-SPs) of Shigella dysenteriae type 1, S. flexneri type 2a, and S. sonnei were covalently bound to carrier proteins. Alone, the O-SPs were not immunogenic in mice. Conjugates of these O-SPs, injected into young outbred mice subcutaneously as saline solutions containing 2.5 micrograms of saccharide, elicited serum IgG and IgM antibodies with booster responses; adsorption onto alum enhanced their immunogenicity. Injection of 25 micrograms of these conjugates into adult volunteers elicited mild local reactions only. Each conjugate induced a significant rise of the geometric mean serum IgG, IgM, and IgA LPS antibody levels. A second injection 6 weeks later did not elicit booster responses, and adsorption of the conjugates onto alum did not enhance their immunogenicity. Conjugate-induced levels of IgA, but not IgG or IgM, declined to preimmunization levels at day 56. The levels of postimmunization antibodies of the three immunoglobulin classes were similar to or higher than those of recruits in the Israel Defense Force following shigellosis caused by S. flexneri type 2a or S. sonnei. These data provide the basis for evaluating these conjugates to prevent shigellosis.

Full text

PDF
3678

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Binns M. M., Vaughan S., Timmis K. N. 'O'-antigens are essential virulence factors of Shigella sonnei and Shigella dysenteriae 1. Zentralbl Bakteriol Mikrobiol Hyg B. 1985 Jun;181(1-2):197–205. [PubMed] [Google Scholar]
  2. Chu C. Y., Liu B. K., Watson D., Szu S. S., Bryla D., Shiloach J., Schneerson R., Robbins J. B. Preparation, characterization, and immunogenicity of conjugates composed of the O-specific polysaccharide of Shigella dysenteriae type 1 (Shiga's bacillus) bound to tetanus toxoid. Infect Immun. 1991 Dec;59(12):4450–4458. doi: 10.1128/iai.59.12.4450-4458.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Cohen D., Green M. S., Block C., Rouach T., Ofek I. Serum antibodies to lipopolysaccharide and natural immunity to shigellosis in an Israeli military population. J Infect Dis. 1988 May;157(5):1068–1071. doi: 10.1093/infdis/157.5.1068. [DOI] [PubMed] [Google Scholar]
  5. Cohen D., Green M. S., Block C., Slepon R., Lerman Y. Natural immunity to shigellosis in two groups with different previous risks of exposure to Shigella is only partly expressed by serum antibodies to lipopolysaccharide. J Infect Dis. 1992 Apr;165(4):785–787. doi: 10.1093/infdis/165.4.785. [DOI] [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. DeMaria A., Jr, Johns M. A., Berberich H., McCabe W. R. Immunization with rough mutants of Salmonella minnesota: initial studies in human subjects. J Infect Dis. 1988 Aug;158(2):301–311. doi: 10.1093/infdis/158.2.301. [DOI] [PubMed] [Google Scholar]
  8. Dmitriev B. A., Backinowsky L. V., Knirel Y. A., Kochetkov N. K. Somatic antigens of Shigella. The strucuture of the specific polysaccharide chain of Shigella dysenteriae type 5 lipopolysaccharide. Eur J Biochem. 1977 Sep;78(2):381–387. doi: 10.1111/j.1432-1033.1977.tb11750.x. [DOI] [PubMed] [Google Scholar]
  9. Douglas C. M., Collier R. J. Exotoxin A of Pseudomonas aeruginosa: substitution of glutamic acid 553 with aspartic acid drastically reduces toxicity and enzymatic activity. J Bacteriol. 1987 Nov;169(11):4967–4971. doi: 10.1128/jb.169.11.4967-4971.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. DuPont H. L., Hornick R. B., Snyder M. J., Libonati J. P., Formal S. B., Gangarosa E. J. Immunity in shigellosis. II. Protection induced by oral live vaccine or primary infection. J Infect Dis. 1972 Jan;125(1):12–16. doi: 10.1093/infdis/125.1.12. [DOI] [PubMed] [Google Scholar]
  11. DuPont H. L., Levine M. M., Hornick R. B., Formal S. B. Inoculum size in shigellosis and implications for expected mode of transmission. J Infect Dis. 1989 Jun;159(6):1126–1128. doi: 10.1093/infdis/159.6.1126. [DOI] [PubMed] [Google Scholar]
  12. Fass R., van de Walle M., Shiloach A., Joslyn A., Kaufman J., Shiloach J. Use of high density cultures of Escherichia coli for high level production of recombinant Pseudomonas aeruginosa exotoxin A. Appl Microbiol Biotechnol. 1991 Oct;36(1):65–69. doi: 10.1007/BF00164700. [DOI] [PubMed] [Google Scholar]
  13. Fattom A., Schneerson R., Watson D. C., Karakawa W. W., Fitzgerald D., Pastan I., Li X., Shiloach J., Bryla D. A., Robbins J. B. Laboratory and clinical evaluation of conjugate vaccines composed of Staphylococcus aureus type 5 and type 8 capsular polysaccharides bound to Pseudomonas aeruginosa recombinant exoprotein A. Infect Immun. 1993 Mar;61(3):1023–1032. doi: 10.1128/iai.61.3.1023-1032.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ferguson W. W., Henderson N. D. Description of Strain C27: A Motile Organism with the Major Antigen of Shigella sonnei Phase I. J Bacteriol. 1947 Aug;54(2):179–181. doi: 10.1128/jb.54.2.179-181.1947. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ferreccio C., Prado V., Ojeda A., Cayyazo M., Abrego P., Guers L., Levine M. M. Epidemiologic patterns of acute diarrhea and endemic Shigella infections in children in a poor periurban setting in Santiago, Chile. Am J Epidemiol. 1991 Sep 15;134(6):614–627. doi: 10.1093/oxfordjournals.aje.a116134. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Forman D. W., Tong M. J., Murrell K. D., Cross J. H. Etiologic study of diarrheal disease in Vietnam. Am J Trop Med Hyg. 1971 Jul;20(4):598–601. doi: 10.4269/ajtmh.1971.20.598. [DOI] [PubMed] [Google Scholar]
  18. Green M. S., Block C., Cohen D., Slater P. E. Four decades of shigellosis in Israel: epidemiology of a growing public health problem. Rev Infect Dis. 1991 Mar-Apr;13(2):248–253. doi: 10.1093/clinids/13.2.248. [DOI] [PubMed] [Google Scholar]
  19. Gupta R. K., Szu S. C., Finkelstein R. A., Robbins J. B. Synthesis, characterization, and some immunological properties of conjugates composed of the detoxified lipopolysaccharide of Vibrio cholerae O1 serotype Inaba bound to cholera toxin. Infect Immun. 1992 Aug;60(8):3201–3208. doi: 10.1128/iai.60.8.3201-3208.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hale T. L., Formal S. B. Oral shigella vaccines. Curr Top Microbiol Immunol. 1989;146:205–211. doi: 10.1007/978-3-642-74529-4_22. [DOI] [PubMed] [Google Scholar]
  21. Hale T. L. Genetic basis of virulence in Shigella species. Microbiol Rev. 1991 Jun;55(2):206–224. doi: 10.1128/mr.55.2.206-224.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Henry F. J., Alam N., Aziz K. M., Rahaman M. M. Dysentery, not watery diarrhoea, is associated with stunting in Bangladeshi children. Hum Nutr Clin Nutr. 1987 Jul;41(4):243–249. [PubMed] [Google Scholar]
  23. Herrington D. A., Van de Verg L., Formal S. B., Hale T. L., Tall B. D., Cryz S. J., Tramont E. C., Levine M. M. Studies in volunteers to evaluate candidate Shigella vaccines: further experience with a bivalent Salmonella typhi-Shigella sonnei vaccine and protection conferred by previous Shigella sonnei disease. Vaccine. 1990 Aug;8(4):353–357. doi: 10.1016/0264-410x(90)90094-3. [DOI] [PubMed] [Google Scholar]
  24. Hochstein H. D., Mills D. F., Outschoorn A. S., Rastogi S. C. The processing and collaborative assay of a reference endotoxin. J Biol Stand. 1983 Oct;11(4):251–260. doi: 10.1016/s0092-1157(83)80013-4. [DOI] [PubMed] [Google Scholar]
  25. Kenne L., Lindberg B., Petersson K., Katzenellenbogen E., Romanowska E. Structural studies of Shigella flexneri O-antigens. Eur J Biochem. 1978 Nov 2;91(1):279–284. doi: 10.1111/j.1432-1033.1978.tb20963.x. [DOI] [PubMed] [Google Scholar]
  26. Kopecko D. J., Washington O., Formal S. B. Genetic and physical evidence for plasmid control of Shigella sonnei form I cell surface antigen. Infect Immun. 1980 Jul;29(1):207–214. doi: 10.1128/iai.29.1.207-214.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. LANDY M., GAINES S., SEAL J. R., WHITSIDE J. E. Antibody responses of man to three types of antityphoid immunizing agents: heat-phenol fluid vaccine, acetone-dehydrated vaccine, and isolated Vi and O antigens. Am J Public Health Nations Health. 1954 Dec;44(12):1572–1579. doi: 10.2105/ajph.44.12.1572. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Leibovici L., Yahav J., Mates A., Linn S., Danon Y. L. Causes of diarrheal disease in soldiers stationed in Lebanon and in northern Israel. Isr J Med Sci. 1984 Apr;20(4):364–365. [PubMed] [Google Scholar]
  29. Lue C., Prince S. J., Fattom A., Schneerson R., Robbins J. B., Mestecky J. Antibody-secreting peripheral blood lymphocytes induced by immunization with a conjugate consisting of Streptococcus pneumoniae type 12F polysaccharide and diphtheria toxoid. Infect Immun. 1990 Aug;58(8):2547–2554. doi: 10.1128/iai.58.8.2547-2554.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Mel D., Gangarosa E. J., Radovanovic M. L., Arsic B. L., Litvinjenko S. Studies on vaccination against bacillary dysentery. 6. Protection of children by oral immunization with streptomycin-dependent Shigella strains. Bull World Health Organ. 1971;45(4):457–464. [PMC free article] [PubMed] [Google Scholar]
  31. Pierce N. F., Reynolds H. Y. Immunity to experimental cholera. I. Protective effect of humoral IgG antitoxin demonstrated by passive immunization. J Immunol. 1974 Sep;113(3):1017–1023. [PubMed] [Google Scholar]
  32. Rauss K., Kontrohr T., Vertényi A., Szendrei L. Serological and chemical studies of Sh. sonnei, Pseudomonas shigelloides and C27 strains. Acta Microbiol Acad Sci Hung. 1970;17(2):157–166. [PubMed] [Google Scholar]
  33. Robbins J. B., Chu C., Schneerson R. Hypothesis for vaccine development: protective immunity to enteric diseases caused by nontyphoidal salmonellae and shigellae may be conferred by serum IgG antibodies to the O-specific polysaccharide of their lipopolysaccharides. Clin Infect Dis. 1992 Aug;15(2):346–361. doi: 10.1093/clinids/15.2.346. [DOI] [PubMed] [Google Scholar]
  34. Robbins J. B., Schneerson R. Polysaccharide-protein conjugates: a new generation of vaccines. J Infect Dis. 1990 May;161(5):821–832. doi: 10.1093/infdis/161.5.821. [DOI] [PubMed] [Google Scholar]
  35. Ronsmans C., Bennish M. L., Wierzba T. Diagnosis and management of dysentery by community health workers. Lancet. 1988 Sep 3;2(8610):552–555. doi: 10.1016/s0140-6736(88)92669-4. [DOI] [PubMed] [Google Scholar]
  36. Schwartzer T. A., Alcid D. V., Numsuwan V., Gocke D. J. Characterization of the human antibody response to an Escherichia coli O111:B4 (J5) vaccine. J Infect Dis. 1988 Nov;158(5):1135–1136. doi: 10.1093/infdis/158.5.1135. [DOI] [PubMed] [Google Scholar]
  37. Sturm S., Jann B., Jann K., Fortnagel P., Timmis K. N. Genetic and biochemical analysis of Shigella dysenteriae 1 O antigen polysaccharide biosynthesis in Escherichia coli K-12: structure and functions of the rfb gene cluster. Microb Pathog. 1986 Jun;1(3):307–324. doi: 10.1016/0882-4010(86)90056-2. [DOI] [PubMed] [Google Scholar]
  38. Taylor D. N., Echeverria P., Pál T., Sethabutr O., Saiborisuth S., Sricharmorn S., Rowe B., Cross J. The role of Shigella spp., enteroinvasive Escherichia coli, and other enteropathogens as causes of childhood dysentery in Thailand. J Infect Dis. 1986 Jun;153(6):1132–1138. doi: 10.1093/infdis/153.6.1132. [DOI] [PubMed] [Google Scholar]
  39. Tsai C. M., Mocca L. F., Frasch C. E. Immunotype epitopes of Neisseria meningitidis lipooligosaccharide types 1 through 8. Infect Immun. 1987 Jul;55(7):1652–1656. doi: 10.1128/iai.55.7.1652-1656.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Watson D. C., Robbins J. B., Szu S. C. Protection of mice against Salmonella typhimurium with an O-specific polysaccharide-protein conjugate vaccine. Infect Immun. 1992 Nov;60(11):4679–4686. doi: 10.1128/iai.60.11.4679-4686.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES