Broad spectrum pyrogenic antagonists and vaccines directed against pyrogenic exotoxins

Abstract

The invention relates to peptides comprising an amino acid sequence homologous to the amino sequence of a fragment of a pyrogenic exotoxin, and functional derivatives of said peptides, capable of eliciting protective immunity against toxic shock induced by a pyrogenic exotoxin or by a mixture of pyrogenic exotoxins. Preferred peptides comprise an amino acid sequence homologous to the amino acid sequence of a fragment of Staphylococcal aureus enterotoxin B (SEB). The invention also relates to broad spectrum pharmaceutical compositions for the treatment, protection against or short-term prophylaxis of toxin-mediated activation of T cells, comprising as active ingredient at least one peptide according to the invention or derivative thereof, and to broad spectrum vaccines for conferring long-term immunity against toxic shock induced by at least one pyrogenic exotoxin are provided. The vaccines comprise as active ingredient at least one peptide according to the invention, or derivative thereof.

1 . A method of inhibiting pyrogenic exotoxin-mediated activation of T-lymphocytes and protecting against toxic shock induced by a pyrogenic exotoxin or by a mixture of pyrogenic exotoxins, in a patient in need of such treatment comprising administering to said patient an inhibitory effective amount of an isolated and purified peptide or of a composition comprising the same, which peptide having an amino acid sequence homologous to an amino acid sequence of a domain of a pyrogenic exotoxin which domain forms a central turn in the exotoxin starting within β-strand 7 and connecting the β-strand 7, via short β-strand 8, to α-helix 4, and ending within α-helix 4, based on the domain numbering of SEB, wherein said peptide does not have toxin agonist activity.  
     
     
         2 . The method according to  claim 1 , wherein said peptide is selected from the group consisting of a peptide having the amino acid sequence of any one of SEQ ID NO:1 to SEQ ID NO:11 and SEQ ID NO:13 to SEQ ID NO:15 and any mixture thereof.  
     
     
         3 . The method according to  claim 1 , wherein said peptide is a peptide having the amino acid sequence according to SEQ ID NO:2.  
     
     
         4 . The method according to  claim 1 , wherein said peptide is a peptide having the amino acid sequence according to SEQ ID NO:13.  
     
     
         5 . The method according to  claim 1 , wherein said peptide is a peptide having an amino acid sequence Xaa (n) KXaa (8) D (SEQ ID NO: 54), wherein Xaa is an amino acid and n is zero or an integer of from 1 to 10, and wherein said peptide does not have toxin agonist activity.  
     
     
         6 . The method according to  claim 1 , wherein said peptide is a peptide having an amino acid sequence Xaa (n) KXaa (3) TXaaQEXaaD (SEQ ID NO:55), wherein Xaa is an amino acid and n is zero or an integer of from 1 to 10, and wherein said peptide does not have toxin agonist activity.  
     
     
         7 . The method according to  claim 1 , wherein said peptide is a peptide having an amino acid sequence Xaa (n) KKXaa (6) LD (SEQ ID NO:56), wherein Xaa is an amino acid and n is zero or an integer of from 1 to 10, and wherein said peptide does not have toxin agonist activity.  
     
     
         8 . The method according to  claim 1 , wherein said peptide is a peptide having an amino acid sequence Ch.aa-Xaa (2) -Hb.aa-Xaa-Hb.aa-Po.aa-Po.aa-Hb.aa-D (SEQ ID NO:57), wherein Xaa is an amino acid, Ch.aa is a charged amino acid, Hb.aa is a hydrophobic amino acid and Po.aa is a polar amino acid and wherein said peptide does not have toxin agonist activity.  
     
     
         9 . A method of eliciting protective immunity against a toxic shock induced by a pyrogenic exotoxin in a patient in need of such treatment comprising administering to said patient an isolated and purified peptide or of an immunogenic composition comprising the same, which peptide having an amino acid sequence homologous to an amino acid sequence of a domain of a pyrogenic exotoxin which domain forms a central turn in the exotoxin starting within β-strand 7 and connecting the β-strand 7, via short β-strand 8, to α-helix 4, and ending within α-helix 4, based on the domain numbering of SEB, wherein said peptide does not have toxin agonist activity.  
     
     
         10 . The method according to  claim 9 , wherein said peptide is selected from a peptide having the amino acid sequence of any one of SEQ ID NO:1 to SEQ ID NO:11 and SEQ ID NO:13 to SEQ ID NO:15 and any mixture thereof.  
     
     
         11 . The method according to  claim 9 , wherein said peptide is a peptide having the amino acid sequence according to SEQ ID NO:2.  
     
     
         12 . The method according to  claim 9 , wherein said peptide is a peptide having the amino acid sequence according to SEQ ID NO:13.  
     
     
         13 . The method according to  claim 9 , wherein said peptide is a peptide having an amino acid sequence Xaa (n) KXaa (8) D (SEQ ID NO: 54), wherein Xaa is an amino acid and n is zero or an integer of from 1 to 10, and wherein said peptide does not have toxin agonist activity.  
     
     
         14 . The method according to  claim 9 , wherein said peptide is a peptide having an amino acid sequence Xaa (n) KXaa (3) TXaaQEXaaD (SEQ ID NO:55), wherein Xaa is an amino acid and n is zero or an integer of from 1 to 10, and wherein said peptide does not have toxin agonist activity.  
     
     
         15 . The method according to  claim 9 , wherein said peptide is a peptide having an amino acid sequence Xaa (n) KKXaa (6) LD (SEQ ID NO:56), wherein Xaa is an amino acid and n is zero or an integer of from 1 to 10, and wherein said peptide does not have toxin agonist activity.  
     
     
         16 . The method according to  claim 9 , wherein said peptide is a peptide having an amino acid sequence Ch.aa-Xaa (2) -Hb.aa-Xaa-Hb.aa-Po.aa-Po.aa-Hb.aa-D (SEQ ID NO:57), wherein Xaa is an amino acid, Ch.aa is a charged amino acid, Hb.aa is a hydrophobic amino acid and Po.aa is a polar amino acid and wherein said peptide does not have toxin agonist activity.  
     
     
         17 . A method of preventing and/or treating incapacitation induced by at least one pyrogenic exotoxin comprising administering to a patient in need of such treatment at least one therapeutically effective dose of an isolated and purified peptide or of a composition comprising the same, which peptide having an amino acid sequence homologous to an amino acid sequence of a domain of a pyrogenic exotoxin which domain forms a central turn in the exotoxin starting within β-strand 7 and connecting the β-strand 7, via short β-strand 8, to α-helix 4, and ending within α-helix 4, based on the domain numbering of SEB, wherein said peptide does not have toxin agonist activity.  
     
     
         18 . The method according to  claim 17 , wherein said peptide is selected from a peptide having the amino acid sequence of any one of SEQ ID NO:1 to SEQ ID NO:11 and SEQ ID NO:13 to SEQ ID NO:15 and any mixture thereof.  
     
     
         19 . The method according to  claim 17 , wherein said peptide is a peptide having the amino acid sequence according to SEQ ID NO:2.  
     
     
         20 . The method according to  claim 17 , wherein said peptide is a peptide having the amino acid sequence according to SEQ ID NO:13.  
     
     
         21 . The method according to  claim 17 , wherein said peptide is a peptide having an amino acid sequence Xaa (n) KXaa (8) D (SEQ ID NO: 54), wherein Xaa is an amino acid and n is zero or an integer of from 1 to 10, and wherein said peptide does not have toxin agonist activity.  
     
     
         22 . The method according to  claim 17 , wherein said peptide is a peptide having an amino acid sequence Xaa (n) KXaa (3) TXaaQEXaaD (SEQ ID NO:55), wherein Xaa is an amino acid and n is zero or an integer of from 1 to 10, and wherein said peptide does not have toxin agonist activity.  
     
     
         23 . The method according to  claim 17 , wherein said peptide is a peptide having an amino acid sequence Xaa (n) KKXaa (6) LD (SEQ ID NO:56), wherein Xaa is an amino acid and n is zero or an integer of from 1 to 10, and wherein said peptide does not have toxin agonist activity.  
     
     
         24 . The method according to  claim 17 , wherein said peptide is a peptide having an amino acid sequence Ch.aa-Xaa (2) -Hb.aa-Xaa-Hb.aa-Po.aa-Po.aa-Hb.aa-D (SEQ ID NO:57), wherein Xaa is an amino acid, Ch.aa is a charged amino acid, Hb.aa is a hydrophobic amino acid and Po.aa is a polar amino acid and wherein said peptide does not have toxin agonist activity.  
     
     
         25 . The method according to any  claim 17 , wherein the effective does of said isolated and purified peptide or composition comprising the same is administered to said patient repeatedly, at predetermined periods of time.  
     
     
         26 . Antibodies directed against a peptide having an amino acid sequence homologous to an amino acid sequence of a domain of a pyrogenic exotoxin, which domain forms a central turn in the exotoxin starting within β-strand 7 and connecting the β-strand 7, via short β-strand 8, to α-helix 4, and ending within α-helix 4, based on the domain numbering of SEB, wherein said peptide does not have toxin agonist activity.  
     
     
         27 . An antiserum containing antibodies directed against a peptide having an amino acid sequence homologous to an amino acid sequence of a domain of a pyrogenic exotoxin, which domain forms a central turn in the exotoxin starting within β-strand 7 and connecting the β-strand 7, via short β-strand 8, to α-helix 4, and ending within α-helix 4, based on the domain numbering of SEB, wherein said peptide does not have toxin agonist activity, which peptide is capable of eliciting the production of said antibodies, preferably in the presence of a suitable immunization adjuvant.  
     
     
         28 . An antiserum according to  claim 27  wherein said suitable immunizing adjuvant is proteosome, KLH or alum or combinations thereof.  
     
     
         29 . An antiserum according to  claim 28  wherein said immunizing adjuvant is a combination of proteosomes and alum or is a combination of KLH and alum.  
     
     
         30 . An antiserum according to  claim 27  which is a domestic animal antiserum.  
     
     
         31 . An antiserum according to  claim 27  capable of alleviating harmful effects and toxic shock induced by a pyrogenic exotoxin or by a mixture of pyrogenic exotoxins.  
     
     
         32 . A method for assessing the efficacy of a vaccine for conferring immunity against one or more pyrogenic toxins comprising determining the ability of serum from an immunized individual to antagonize toxin-mediated activation of T cells.  
     
     
         33 . A method according to  claim 32  wherein the ability of serum from an immunized individual to antagonize toxin-mediated activation of T cells is determined by measuring the inhibition of expression of pyrogenic toxin-induced mRNA encoded by the IL-2, IFN-γ or TNF-β genes.  
     
     
         34 . A kit for assessing the efficacy of a vaccine for conferring immunity against one or more pyrogenic toxins comprising determining the ability of serum from an immunized individual to antagonize toxin-mediated activation of T cells by the method of claim  47 .