Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1997 Jun;65(6):2402–2412. doi: 10.1128/iai.65.6.2402-2412.1997

Expression and immunogenicity of an Echinococcus granulosus fatty acid-binding protein in live attenuated Salmonella vaccine strains.

J A Chabalgoity 1, J A Harrison 1, A Esteves 1, R Demarco de Hormaeche 1, R Ehrlich 1, C M Khan 1, C E Hormaeche 1
PMCID: PMC175333  PMID: 9169781

Abstract

Fatty acid-binding proteins (FABPs) are candidate molecules for vaccines against several parasitic platyhelminths. A FABP from the cestode Echinococcus granulosus (EgDf1) was expressed in Salmonella vaccine strains as a C-terminal fusion to fragment C of tetanus toxin (TetC) by using expression vector pTECH. The fusion protein was equally expressed in several attenuated vaccine strains derived from bacteria with different genetic backgrounds and different attenuating mutations. Single-dose immunization experiments with the aroA Salmonella typhimurium strain SL3261 carrying the pTECH-EgDf1 construct were conducted with mice, using both the intravenous and the oral routes. Surprisingly, the antibody response to EgDf1 and the antigen-specific cytokine production in spleen cells were stronger in mice immunized orally. Furthermore, immune mouse sera strongly reacted with fixed sections of the worm's larval stage. Analysis of the isotype distribution of the specific anti-EgDf1 antibodies showed similar production of immunoglobulin G1 (IgG1) and IgG2a together with specific IgA antibodies. In addition, stimulation of spleen cells from mice immunized with the different constructs with either Salmonella lysate, TetC, or EgDf1 showed that, together with Th1-related cytokines (gamma interferon and interleukin 2 [IL-2]), significant levels of a Th2 cytokine (IL-5) were produced specifically, indicating a Th2 component to the response to the Salmonella carrier and to the recombinant antigens. Salmonellae expressing the TetC-rEgDfl fusion are currently under evaluation as potential vaccines against E. granulosus.

Full Text

The Full Text of this article is available as a PDF (732.5 KB).

Selected References

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

  1. Brett S. J., Dunlop L., Liew F. Y., Tite J. P. Influence of the antigen delivery system on immunoglobulin isotype selection and cytokine production in response to influenza A nucleoprotein. Immunology. 1993 Oct;80(2):306–312. [PMC free article] [PubMed] [Google Scholar]
  2. Brown A., Hormaeche C. E., Demarco de Hormaeche R., Winther M., Dougan G., Maskell D. J., Stocker B. A. An attenuated aroA Salmonella typhimurium vaccine elicits humoral and cellular immunity to cloned beta-galactosidase in mice. J Infect Dis. 1987 Jan;155(1):86–92. doi: 10.1093/infdis/155.1.86. [DOI] [PubMed] [Google Scholar]
  3. Chabalgoity J. A., Khan C. M., Nash A. A., Hormaeche C. E. A Salmonella typhimurium htrA live vaccine expressing multiple copies of a peptide comprising amino acids 8-23 of herpes simplex virus glycoprotein D as a genetic fusion to tetanus toxin fragment C protects mice from herpes simplex virus infection. Mol Microbiol. 1996 Feb;19(4):791–801. doi: 10.1046/j.1365-2958.1996.426965.x. [DOI] [PubMed] [Google Scholar]
  4. Chabalgoity J. A., Villareal-Ramos B., Khan C. M., Chatfield S. N., de Hormaeche R. D., Hormaeche C. E. Influence of preimmunization with tetanus toxoid on immune responses to tetanus toxin fragment C-guest antigen fusions in a Salmonella vaccine carrier. Infect Immun. 1995 Jul;63(7):2564–2569. doi: 10.1128/iai.63.7.2564-2569.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Coffman R. L., Seymour B. W., Lebman D. A., Hiraki D. D., Christiansen J. A., Shrader B., Cherwinski H. M., Savelkoul H. F., Finkelman F. D., Bond M. W. The role of helper T cell products in mouse B cell differentiation and isotype regulation. Immunol Rev. 1988 Feb;102:5–28. doi: 10.1111/j.1600-065x.1988.tb00739.x. [DOI] [PubMed] [Google Scholar]
  6. Coulson N. M., Fulop M., Titball R. W. Bacillus anthracis protective antigen, expressed in Salmonella typhimurium SL 3261, affords protection against anthrax spore challenge. Vaccine. 1994 Nov;12(15):1395–1401. doi: 10.1016/0264-410x(94)90148-1. [DOI] [PubMed] [Google Scholar]
  7. Craig P. S., Liu D., Deshan L., Ding Z., Zhaoxun D. Hydatid disease in China. Parasitol Today. 1991 Feb;7(2):46–50. doi: 10.1016/0169-4758(91)90188-t. [DOI] [PubMed] [Google Scholar]
  8. DeKruyff R. H., Mosmann R. R., Umetsu D. T. Induction of antibody synthesis by CD4+ T cells: IL 5 is essential for induction of antigen-specific antibody responses by TH2 but not TH1 clones. Eur J Immunol. 1990 Oct;20(10):2219–2227. doi: 10.1002/eji.1830201010. [DOI] [PubMed] [Google Scholar]
  9. Esteves A., Dallagiovanna B., Ehrlich R. A developmentally regulated gene of Echinococcus granulosus codes for a 15.5-kilodalton polypeptide related to fatty acid binding proteins. Mol Biochem Parasitol. 1993 Apr;58(2):215–222. doi: 10.1016/0166-6851(93)90043-w. [DOI] [PubMed] [Google Scholar]
  10. Finkelman F. D., Katona I. M., Mosmann T. R., Coffman R. L. IFN-gamma regulates the isotypes of Ig secreted during in vivo humoral immune responses. J Immunol. 1988 Feb 15;140(4):1022–1027. [PubMed] [Google Scholar]
  11. Hayes L. J., Conlan J. W., Everson J. S., Ward M. E., Clarke I. N. Chlamydia trachomatis major outer membrane protein epitopes expressed as fusions with LamB in an attenuated aro A strain of Salmonella typhimurium; their application as potential immunogens. J Gen Microbiol. 1991 Jul;137(7):1557–1564. doi: 10.1099/00221287-137-7-1557. [DOI] [PubMed] [Google Scholar]
  12. Hillyer G. V., García Rosa M. I., Alicea H., Hernandez A. Successful vaccination against murine Schistosoma mansoni infection with a purified 12 Kd Fasciola hepatica cross-reactive antigen. Am J Trop Med Hyg. 1988 Jan;38(1):103–110. doi: 10.4269/ajtmh.1988.38.103. [DOI] [PubMed] [Google Scholar]
  13. Jones B. D., Falkow S. Salmonellosis: host immune responses and bacterial virulence determinants. Annu Rev Immunol. 1996;14:533–561. doi: 10.1146/annurev.immunol.14.1.533. [DOI] [PubMed] [Google Scholar]
  14. Khan C. M., Villarreal-Ramos B., Pierce R. J., Demarco de Hormaeche R., McNeill H., Ali T., Chatfield S., Capron A., Dougan G., Hormaeche C. E. Construction, expression, and immunogenicity of multiple tandem copies of the Schistosoma mansoni peptide 115-131 of the P28 glutathione S-transferase expressed as C-terminal fusions to tetanus toxin fragment C in a live aro-attenuated vaccine strain of Salmonella. J Immunol. 1994 Dec 15;153(12):5634–5642. [PubMed] [Google Scholar]
  15. Khan C. M., Villarreal-Ramos B., Pierce R. J., Riveau G., Demarco de Hormaeche R., McNeill H., Ali T., Fairweather N., Chatfield S., Capron A. Construction, expression, and immunogenicity of the Schistosoma mansoni P28 glutathione S-transferase as a genetic fusion to tetanus toxin fragment C in a live Aro attenuated vaccine strain of Salmonella. Proc Natl Acad Sci U S A. 1994 Nov 8;91(23):11261–11265. doi: 10.1073/pnas.91.23.11261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lundberg K. S., Shoemaker D. D., Adams M. W., Short J. M., Sorge J. A., Mathur E. J. High-fidelity amplification using a thermostable DNA polymerase isolated from Pyrococcus furiosus. Gene. 1991 Dec 1;108(1):1–6. doi: 10.1016/0378-1119(91)90480-y. [DOI] [PubMed] [Google Scholar]
  17. Maizels R. M., Bundy D. A., Selkirk M. E., Smith D. F., Anderson R. M. Immunological modulation and evasion by helminth parasites in human populations. Nature. 1993 Oct 28;365(6449):797–805. doi: 10.1038/365797a0. [DOI] [PubMed] [Google Scholar]
  18. Maskell D. J., Sweeney K. J., O'Callaghan D., Hormaeche C. E., Liew F. Y., Dougan G. Salmonella typhimurium aroA mutants as carriers of the Escherichia coli heat-labile enterotoxin B subunit to the murine secretory and systemic immune systems. Microb Pathog. 1987 Mar;2(3):211–221. doi: 10.1016/0882-4010(87)90022-2. [DOI] [PubMed] [Google Scholar]
  19. Mosmann T. R., Cherwinski H., Bond M. W., Giedlin M. A., Coffman R. L. Two types of murine helper T cell clone. I. Definition according to profiles of lymphokine activities and secreted proteins. J Immunol. 1986 Apr 1;136(7):2348–2357. [PubMed] [Google Scholar]
  20. Murray P. D., McKenzie D. T., Swain S. L., Kagnoff M. F. Interleukin 5 and interleukin 4 produced by Peyer's patch T cells selectively enhance immunoglobulin A expression. J Immunol. 1987 Oct 15;139(8):2669–2674. [PubMed] [Google Scholar]
  21. Ramarathinam L., Shaban R. A., Niesel D. W., Klimpel G. R. Interferon gamma (IFN-gamma) production by gut-associated lymphoid tissue and spleen following oral Salmonella typhimurium challenge. Microb Pathog. 1991 Nov;11(5):347–356. doi: 10.1016/0882-4010(91)90020-b. [DOI] [PubMed] [Google Scholar]
  22. Schödel F., Milich D. R., Will H. Hepatitis B virus nucleocapsid/pre-S2 fusion proteins expressed in attenuated Salmonella for oral vaccination. J Immunol. 1990 Dec 15;145(12):4317–4321. [PubMed] [Google Scholar]
  23. Simonet M., Fortineau N., Beretti J. L., Berche P. Immunization with live aroA recombinant Salmonella typhimurium producing invasin inhibits intestinal translocation of Yersinia pseudotuberculosis. Infect Immun. 1994 Mar;62(3):863–867. doi: 10.1128/iai.62.3.863-867.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Smith B. P., Reina-Guerra M., Hoiseth S. K., Stocker B. A., Habasha F., Johnson E., Merritt F. Aromatic-dependent Salmonella typhimurium as modified live vaccines for calves. Am J Vet Res. 1984 Jan;45(1):59–66. [PubMed] [Google Scholar]
  25. Stabel T. J., Mayfield J. E., Tabatabai L. B., Wannemuehler M. J. Swine immunity to an attenuated Salmonella typhimurium mutant containing a recombinant plasmid which codes for production of a 31-kilodalton protein of Brucella abortus. Infect Immun. 1991 Sep;59(9):2941–2947. doi: 10.1128/iai.59.9.2941-2947.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Tendler M., Vilar M. M., Brito C. A., Freire N. M., Katz N., Simpson A. Vaccination against schistosomiasis and fascioliasis with the new recombinant antigen Sm14: potential basis of a multi-valent anti-helminth vaccine? Mem Inst Oswaldo Cruz. 1995 Mar-Apr;90(2):255–256. doi: 10.1590/s0074-02761995000200022. [DOI] [PubMed] [Google Scholar]
  27. Tijhaar E. J., Zheng-Xin Y., Karlas J. A., Meyer T. F., Stukart M. J., Osterhaus A. D., Mooi F. R. Construction and evaluation of an expression vector allowing the stable expression of foreign antigens in a Salmonella typhimurium vaccine strain. Vaccine. 1994 Aug;12(11):1004–1011. doi: 10.1016/0264-410x(94)90336-0. [DOI] [PubMed] [Google Scholar]
  28. Tonkonogy S. L., McKenzie D. T., Swain S. L. Regulation of isotype production by IL-4 and IL-5. Effects of lymphokines on Ig production depend on the state of activation of the responding B cells. J Immunol. 1989 Jun 15;142(12):4351–4360. [PubMed] [Google Scholar]
  29. VanCott J. L., Staats H. F., Pascual D. W., Roberts M., Chatfield S. N., Yamamoto M., Coste M., Carter P. B., Kiyono H., McGhee J. R. Regulation of mucosal and systemic antibody responses by T helper cell subsets, macrophages, and derived cytokines following oral immunization with live recombinant Salmonella. J Immunol. 1996 Feb 15;156(4):1504–1514. [PubMed] [Google Scholar]
  30. Villarreal B., Mastroeni P., de Hormaeche R. D., Hormaeche C. E. Proliferative and T-cell specific interleukin (IL-2/IL-4) production responses in spleen cells from mice vaccinated with aroA live attenuated Salmonella vaccines. Microb Pathog. 1992 Oct;13(4):305–315. doi: 10.1016/0882-4010(92)90040-u. [DOI] [PubMed] [Google Scholar]
  31. Xu D., McSorley S. J., Chatfield S. N., Dougan G., Liew F. Y. Protection against Leishmania major infection in genetically susceptible BALB/c mice by gp63 delivered orally in attenuated Salmonella typhimurium (AroA- AroD-). Immunology. 1995 May;85(1):1–7. [PMC free article] [PubMed] [Google Scholar]
  32. Yang D. M., Fairweather N., Button L. L., McMaster W. R., Kahl L. P., Liew F. Y. Oral Salmonella typhimurium (AroA-) vaccine expressing a major leishmanial surface protein (gp63) preferentially induces T helper 1 cells and protective immunity against leishmaniasis. J Immunol. 1990 Oct 1;145(7):2281–2285. [PubMed] [Google Scholar]

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

RESOURCES