Immunogenicity of genetically engineered glutathione S-transferase fusion proteins containing a T-cell epitope from diphtheria toxin.
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.
Pillai
S
S
Department of Immunology, Lederle-Praxis Biologicals Division, West Henrietta, New York 14586.
Dermody
K
K
Metcalf
B
B
D000595:Amino Acid Sequence; D000818:Animals; D000917:Antibody Formation; D000947:Antigens, Helminth / Q000276:immunology*; D001483:Base Sequence; D004167:Diphtheria Toxin / Q000276:immunology*; D000939:Epitopes; D005260:Female; D005982:Glutathione Transferase / Q000276:immunology*; D007156:Immunologic Memory; D008213:Lymphocyte Activation; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000276:immunology*; D012549:Schistosoma japonicum / Q000201:enzymology / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology
D000947:Antigens, Helminth; D004167:Diphtheria Toxin; D000939:Epitopes; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase