Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1995 Apr;63(4):1535–1540. doi: 10.1128/iai.63.4.1535-1540.1995

Immunogenicity of genetically engineered glutathione S-transferase fusion proteins containing a T-cell epitope from diphtheria toxin.

S Pillai 1, K Dermody 1, B Metcalf 1
PMCID: PMC173186  PMID: 7534277

Abstract

Glutathione S-transferase (GST) has been shown to induce a marginal antibody response in experimental animals as well as partial protection against a number of parasitic worms, including Schistosoma and Fasciola species. The objective of our study was to increase the immunogenicity of GST by adding heterologous T-cell epitopes at the carboxy terminus of the protein. We generated recombinant GST proteins by attaching one or three tandem repeats of a T-cell epitope of CRM197, a nontoxic variant of diphtheria toxin. This T-cell epitope encoding the region of amino acids 366 to 383 of CRM197, when contained in a GST fusion protein and/or after purification as a recombinant peptide, retained the ability to induce a CRM197-specific T-cell response. The fusion protein containing a single T-cell epitope induced a strong T-cell proliferative response to GST and also enhanced anti-GST antibody production in mice. The addition of three repeats of the epitope did not augment the responses when compared with the responses of GST itself. The results suggest that the addition of a single T-cell epitope to a larger protein like GST increases the immunogenicity of the protein.

Full Text

The Full Text of this article is available as a PDF (237.8 KB).

Selected References

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

  1. Auriault C., Balloul J. M., Pierce R. J., Damonneville M., Sondermeijer P., Capron A. Helper T cells induced by a purified 28-kilodalton antigen of Schistosoma mansoni protect rats against infection. Infect Immun. 1987 May;55(5):1163–1169. doi: 10.1128/iai.55.5.1163-1169.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Auriault C., Gras-Masse H., Wolowczuk I., Pierce R. J., Balloul J. M., Neyrinck J. L., Drobecq H., Tartar A., Capron A. Analysis of T and B cell epitopes of the Schistosoma mansoni P28 antigen in the rat model by using synthetic peptides. J Immunol. 1988 Sep 1;141(5):1687–1694. [PubMed] [Google Scholar]
  3. Balloul J. M., Grzych J. M., Pierce R. J., Capron A. A purified 28,000 dalton protein from Schistosoma mansoni adult worms protects rats and mice against experimental schistosomiasis. J Immunol. 1987 May 15;138(10):3448–3453. [PubMed] [Google Scholar]
  4. Balloul J. M., Sondermeyer P., Dreyer D., Capron M., Grzych J. M., Pierce R. J., Carvallo D., Lecocq J. P., Capron A. Molecular cloning of a protective antigen of schistosomes. Nature. 1987 Mar 12;326(6109):149–153. doi: 10.1038/326149a0. [DOI] [PubMed] [Google Scholar]
  5. Bixler G. S., Jr, Eby R., Dermody K. M., Woods R. M., Seid R. C., Pillai S. Synthetic peptide representing a T-cell epitope of CRM197 substitutes as carrier molecule in a Haemophilus influenzae type B (Hib) conjugate vaccine. Adv Exp Med Biol. 1989;251:175–180. doi: 10.1007/978-1-4757-2046-4_15. [DOI] [PubMed] [Google Scholar]
  6. Bixler G. S., Jr, Pillai S. The cellular basis of the immune response to conjugate vaccines. Contrib Microbiol Immunol. 1989;10:18–47. [PubMed] [Google Scholar]
  7. Borras-Cuesta F., Petit-Camurdan A., Fedon Y. Engineering of immunogenic peptides by co-linear synthesis of determinants recognized by B and T cells. Eur J Immunol. 1987 Aug;17(8):1213–1215. doi: 10.1002/eji.1830170820. [DOI] [PubMed] [Google Scholar]
  8. Brophy P. M., Pritchard D. I. Parasitic helminth glutathione S-transferases: an update on their potential as targets for immuno- and chemotherapy. Exp Parasitol. 1994 Aug;79(1):89–96. doi: 10.1006/expr.1994.1067. [DOI] [PubMed] [Google Scholar]
  9. Bushara H. O., Bashir M. E., Malik K. H., Mukhtar M. M., Trottein F., Capron A., Taylor M. G. Suppression of Schistosoma bovis egg production in cattle by vaccination with either glutathione S-transferase or keyhole limpet haemocyanin. Parasite Immunol. 1993 Jul;15(7):383–390. doi: 10.1111/j.1365-3024.1993.tb00623.x. [DOI] [PubMed] [Google Scholar]
  10. Cherfas J. New hope for vaccine against schistosomiasis. Science. 1991 Feb 8;251(4994):630–631. doi: 10.1126/science.1992516. [DOI] [PubMed] [Google Scholar]
  11. Francis M. J., Hastings G. Z., Syred A. D., McGinn B., Brown F., Rowlands D. J. Non-responsiveness to a foot-and-mouth disease virus peptide overcome by addition of foreign helper T-cell determinants. Nature. 1987 Nov 12;330(6144):168–170. doi: 10.1038/330168a0. [DOI] [PubMed] [Google Scholar]
  12. Good M. F., Maloy W. L., Lunde M. N., Margalit H., Cornette J. L., Smith G. L., Moss B., Miller L. H., Berzofsky J. A. Construction of synthetic immunogen: use of new T-helper epitope on malaria circumsporozoite protein. Science. 1987 Feb 27;235(4792):1059–1062. doi: 10.1126/science.2434994. [DOI] [PubMed] [Google Scholar]
  13. Grzych J. M., Grezel D., Xu C. B., Neyrinck J. L., Capron M., Ouma J. H., Butterworth A. E., Capron A. IgA antibodies to a protective antigen in human Schistosomiasis mansoni. J Immunol. 1993 Jan 15;150(2):527–535. [PubMed] [Google Scholar]
  14. Hart M. K., Palker T. J., Matthews T. J., Langlois A. J., Lerche N. W., Martin M. E., Scearce R. M., McDanal C., Bolognesi D. P., Haynes B. F. Synthetic peptides containing T and B cell epitopes from human immunodeficiency virus envelope gp120 induce anti-HIV proliferative responses and high titers of neutralizing antibodies in rhesus monkeys. J Immunol. 1990 Oct 15;145(8):2677–2685. [PubMed] [Google Scholar]
  15. Howell M. J., Board P. G., Boray J. C. Glutathione S-transferases in Fasciola hepatica. J Parasitol. 1988 Aug;74(4):715–718. [PubMed] [Google Scholar]
  16. Lightowlers M. W., Mitchell G. F. Assessment of the prevalence and titer of antibodies to a candidate schistosomiasis vaccine molecule, Sj26, in several human serum banks. Acta Trop. 1989 Jul;46(4):229–238. doi: 10.1016/0001-706x(89)90023-5. [DOI] [PubMed] [Google Scholar]
  17. Lise L. D., Mazier D., Jolivet M., Audibert F., Chedid L., Schlesinger D. Enhanced epitopic response to a synthetic human malarial peptide by preimmunization with tetanus toxoid carrier. Infect Immun. 1987 Nov;55(11):2658–2661. doi: 10.1128/iai.55.11.2658-2661.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Löwenadler B., Svennerholm A. M., Gidlund M., Holmgren E., Krook K., Svanholm C., Ulff S., Josephson S. Enhanced immunogenicity of recombinant peptide fusions containing multiple copies of a heterologous T helper epitope. Eur J Immunol. 1990 Jul;20(7):1541–1545. doi: 10.1002/eji.1830200720. [DOI] [PubMed] [Google Scholar]
  19. Mannervik B. The isoenzymes of glutathione transferase. Adv Enzymol Relat Areas Mol Biol. 1985;57:357–417. doi: 10.1002/9780470123034.ch5. [DOI] [PubMed] [Google Scholar]
  20. Milich D. R., McLachlan A., Moriarty A., Thornton G. B. A single 10-residue pre-S(1) peptide can prime T cell help for antibody production to multiple epitopes within the pre-S(1), pre-S(2), and S regions of HBsAg. J Immunol. 1987 Jun 15;138(12):4457–4465. [PubMed] [Google Scholar]
  21. Mitchell G. F., Davern K. M., Wood S. M., Wright M. D., Argyropoulos V. P., McLeod K. S., Tiu W. U., Garcia E. G. Attempts to induce resistance in mice to Schistosoma japonicum and Schistosoma mansoni by exposure to crude schistosome antigens plus cloned glutathione-S-transferases. Immunol Cell Biol. 1990 Dec;68(Pt 6):377–385. doi: 10.1038/icb.1990.51. [DOI] [PubMed] [Google Scholar]
  22. Mitchell G. F., Garcia E. G., Davern K. M., Tiu W. U., Smith D. B. Sensitization against the parasite antigen Sj26 is not sufficient for consistent expression of resistance to Schistosoma japonicum in mice. Trans R Soc Trop Med Hyg. 1988;82(6):885–889. doi: 10.1016/0035-9203(88)90027-2. [DOI] [PubMed] [Google Scholar]
  23. Mitchell G. F. Glutathione S-transferases - potential components of anti-schistosome vaccines? Parasitol Today. 1989 Feb;5(2):34–37. doi: 10.1016/0169-4758(89)90185-3. [DOI] [PubMed] [Google Scholar]
  24. Mitchison N. A. The carrier effect in the secondary response to hapten-protein conjugates. II. Cellular cooperation. Eur J Immunol. 1971 Jan;1(1):18–27. doi: 10.1002/eji.1830010104. [DOI] [PubMed] [Google Scholar]
  25. Pancre V., Wolowczuk I., Guerret S., Copin M. C., Delanoye A., Capron A., Auriault C. Protective effect of rSm28GST-specific T cells in schistosomiasis: role of gamma interferon. Infect Immun. 1994 Sep;62(9):3723–3730. doi: 10.1128/iai.62.9.3723-3730.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schutze M. P., Leclerc C., Jolivet M., Audibert F., Chedid L. Carrier-induced epitopic suppression, a major issue for future synthetic vaccines. J Immunol. 1985 Oct;135(4):2319–2322. [PubMed] [Google Scholar]
  27. Sexton J. L., Milner A. R., Panaccio M., Waddington J., Wijffels G., Chandler D., Thompson C., Wilson L., Spithill T. W., Mitchell G. F. Glutathione S-transferase. Novel vaccine against Fasciola hepatica infection in sheep. J Immunol. 1990 Dec 1;145(11):3905–3910. [PubMed] [Google Scholar]
  28. Sexton J. L., Wilce M. C., Colin T., Wijffels G. L., Salvatore L., Feil S., Parker M. W., Spithill T. W., Morrison C. A. Vaccination of sheep against Fasciola hepatica with glutathione S-transferase. Identification and mapping of antibody epitopes on a three-dimensional model of the antigen. J Immunol. 1994 Feb 15;152(4):1861–1872. [PubMed] [Google Scholar]
  29. Smith D. B., Davern K. M., Board P. G., Tiu W. U., Garcia E. G., Mitchell G. F. Mr 26,000 antigen of Schistosoma japonicum recognized by resistant WEHI 129/J mice is a parasite glutathione S-transferase. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8703–8707. doi: 10.1073/pnas.83.22.8703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Smith D. B., Rubira M. R., Simpson R. J., Davern K. M., Tiu W. U., Board P. G., Mitchell G. F. Expression of an enzymatically active parasite molecule in Escherichia coli: Schistosoma japonicum glutathione S-transferase. Mol Biochem Parasitol. 1988 Jan 15;27(2-3):249–256. doi: 10.1016/0166-6851(88)90044-8. [DOI] [PubMed] [Google Scholar]
  31. Triebel F., Autran B., De Roquefeuil S., Falmagne P., Debré P. Immune response to diphtheria toxin and to different CNBr fragments: evidence for different B and T cell reactivities. Eur J Immunol. 1986 Jan;16(1):47–53. doi: 10.1002/eji.1830160110. [DOI] [PubMed] [Google Scholar]
  32. Varley C. A., Dunne D. W., Havercroft J. C. The influence of adjuvant on humoral responses to glutathione-S-transferase fusion proteins. Parasite Immunol. 1992 Sep;14(5):557–562. doi: 10.1111/j.1365-3024.1992.tb00028.x. [DOI] [PubMed] [Google Scholar]
  33. Watson R. J., Clements J. B. A herpes simplex virus type 1 function continuously required for early and late virus RNA synthesis. Nature. 1980 May 29;285(5763):329–330. doi: 10.1038/285329a0. [DOI] [PubMed] [Google Scholar]
  34. Wolowczuk I., Auriault C., Gras-Masse H., Vendeville C., Balloul J. M., Tartar A., Capron A. Protective immunity in mice vaccinated with the Schistosoma mansoni P-28-1 antigen. J Immunol. 1989 Feb 15;142(4):1342–1350. [PubMed] [Google Scholar]

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

RESOURCES