title: Double mutant and formaldehyde inactivated TSST-1 as vaccine candidates for TSST-1-induced toxic shock syndrome.
abstract: Up to now there is no treatment for staphylococcal toxic shock syndrome, a disease mainly induced by toxic shock syndrome toxin-1(TSST-1). There is great demand in finding means to control the disease, one of them is the development of an effective and safe vaccine against TSST-1. In this study we constructed a series of vaccine candidates and investigated their biological activity, toxicity, and potential to invoke an immune response. TSST-1 was isolated from Stahylococcus aureus supernatants and recombinantly expressed as a N-terminal 6x histidine-tagged protein in Escherichia coli. In order to obtain molecules with minimal toxicity we constructed single mutants (G31R and H135A) and one double mutant (G31R/H135A) with both residues exchanged. We also detoxified native TSST-1 isolated from S. aureus, and recombinantly expressed TSST-1 by treatment with formaldehyde. Functional activity of native and recombinant TSST-1 and grade of inocuity of mutants and toxoids was determined by investigating mitogenity, T-cell activation, and cytokine release upon stimulation of human mononuclear cells with the vaccine candidates. All substances were tested in a rabbit immunization study. After primary immunization and three additional boosts all vaccinated animals developed antibody titers against TSST-1 and were protected against challenge with a lethal doses of superantigen potentiated with lipopolysaccharide.
authors.affiliation: Biomedizinische Forschungsgesellschaft mbH, Schwarzspanierstrasse 15/1/19, A-1090, Vienna, Austria.
authors.forename: Jörg
authors.initials: J
authors.lastname: Gampfer
authors.forename: Vojtech
authors.initials: V
authors.lastname: Thon
authors.forename: Heinz
authors.initials: H
authors.lastname: Gulle
authors.forename: Hermann M
authors.initials: HM
authors.lastname: Wolf
authors.forename: Martha M
authors.initials: MM
authors.lastname: Eibl
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D001427:Bacterial Toxins; D004768:Enterotoxins / Q000276:immunology* / Q000494:pharmacology / Q000633:toxicity; D005260:Female; D005557:Formaldehyde / Q000494:pharmacology; D016209:Interleukin-8 / Q000096:biosynthesis; D008213:Lymphocyte Activation; D008934:Mitogens / Q000494:pharmacology; D009154:Mutation; D011817:Rabbits; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000032:analysis; D012772:Shock, Septic / Q000517:prevention & control*; D013209:Staphylococcal Vaccines / Q000276:immunology*; D018089:Superantigens; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis; D015164:Vaccines, Inactivated / Q000276:immunology; D014614:Vaccines, Synthetic / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D001427:Bacterial Toxins; D004768:Enterotoxins; D016209:Interleukin-8; D008934:Mitogens; D016693:Receptors, Antigen, T-Cell, alpha-beta; D013209:Staphylococcal Vaccines; D018089:Superantigens; D014409:Tumor Necrosis Factor-alpha; D015164:Vaccines, Inactivated; D014614:Vaccines, Synthetic; C030107:enterotoxin F, Staphylococcal; D005557:Formaldehyde