Microbial delivery system

Abstract

The present invention provides methods and compositions for treating or preventing allergic responses, particularly anaphylactic allergic responses, in subjects who are allergic to allergens or susceptible to allergies. Methods of the present invention utilize administration of microorganisms to subjects, where the microorganisms produce allergens and protect the subjects from exposure to the allergens until phagocytosed by antigen-presenting cells. Particularly preferred microorganisms are gram-negative bacteria, gram-positive bacteria, and yeast. Particularly preferred allergens are proteins found in foods, venoms, drugs and latex that elicit allergic reactions and anaphylactic allergic reactions in individuals who are allergic to the proteins or are susceptible to allergies to the proteins. The proteins may also be modified to reduce the ability of the proteins to bind and crosslink IgE antibodies and thereby reduce the risk of eliciting anaphylaxis without affecting T-cell mediated Th1-type immunity.

We claim:  
     
         1 . A method of treating an allergy in a subject susceptible to an anaphylactic allergic response to an allergen, the method comprising steps of: 
 providing a composition comprising microorganisms that produce the allergen; and    administering the composition to the subject at an effective and non-toxic dose.    
     
     
         2 . The method of  claim 1 , wherein in the step of providing, the microorganism is selected from the group consisting of: bacteria, fungi, viruses, algae, and protozoa.  
     
     
         3 . The method of  claim 1 , wherein in the step of providing, the microorganism is selected from the group consisting of: gram-negative bacteria, gram-positive bacteria, and yeast.  
     
     
         4 . The method of  claim 1 , wherein in the step of providing, the microorganism is selected from the group consisting of:  E. coli , Lactococcus, Listeria, Vibrio, Salmonella and  S. cerevisiae.    
     
     
         5 . The method of  claim 1 , wherein in the step of providing, the allergen is found in foods, venoms, or latex.  
     
     
         6 . The method of  claim 1 , wherein in the step of providing, the allergen is a protein found in peanuts, milk, eggs, seafood, nuts, dairy products and fruit.  
     
     
         7 . The method of  claim 1 , wherein in the step of providing, the allergen is a protein found in bee venom.  
     
     
         8 . The method of  claim 1 , wherein in the step of providing, the allergen is Ara h 1, Ara h 2, Ara h 3, or a polypeptide portion thereof.  
     
     
         9 . The method of  claim 1 , wherein in the step of providing, the allergen is protein modified to have a reduced ability to bind and crosslink IgE antibodies.  
     
     
         10 . The method of  claim 1 , wherein in the step of providing, the microorganisms produce a portion of the allergen.  
     
     
         11 . The method of  claim 10 , wherein in the step of providing, the portion of the allergen produced has a reduced number of IgE binding sites as compared to the allergen.  
     
     
         12 . The method of  claim 1 , wherein in the step of providing, the allergen is a polypeptide and production of the allergen is inducible; and wherein after the step of administering, the method further comprises the step of inducing expression of the polypeptide.  
     
     
         13 . The method of  claim 12 , wherein in the step of inducing, the polypeptide is secreted into a periplasm or secreted outside the cell.  
     
     
         14 . The method of  claim 1 , wherein the step of providing comprises providing a composition comprising gram-negative bacteria or yeast that secretes the allergen into a periplasm.  
     
     
         15 . The method of  claim 1 , wherein in the step of providing, the allergen is a small molecule.  
     
     
         16 . A composition comprising a microorganism that produces an allergen that elicits an anaphylactic allergic reaction in a subject allergic to the allergen.  
     
     
         17 . The composition of  claim 16 , wherein the allergen is a polypeptide or small molecule.  
     
     
         18 . The composition of  claim 16 , wherein the microorganism is selected from the group consisting of: bacteria, fungi, viruses, algae, and protozoa.  
     
     
         19 . The composition of  claim 16 , wherein the microorganism is selected from the group consisting of: gram-negative bacteria, gram-positive bacteria, and yeast.  
     
     
         20 . The composition of  claim 16 , wherein the microorganism is selected from the group consisting of:  E. coli , Lactococcus, Listeria, Vibrio, Salmonella and  S. cerevisiae    
     
     
         21 . The composition of  claim 16 , wherein the allergen found in foods, venoms, or latex.  
     
     
         22 . The composition of  claim 16 , wherein the allergen is an allergen found in peanuts, milk, eggs, seafood, nuts, dairy products and fruit.  
     
     
         23 . The composition of  claim 16 , wherein the allergen found in bee venom.  
     
     
         24 . The composition of  claim 16 , wherein the protein is Ara h 1, Ara h 2, Ara h 3, or a polypeptide portion thereof.  
     
     
         25 . The composition of  claim 16 , wherein the allergen is modified to have a reduced ability to bind and crosslink IgE antibodies.  
     
     
         26 . The composition of  claim 16 , wherein the microorganism produces a portion of the allergen.  
     
     
         27 . The composition of  claim 16 , wherein the portion of the allergen produced has a reduced number of IgE binding sites as compared to the allergen.  
     
     
         28 . The composition of  claim 16 , wherein production of the allergen is inducible.  
     
     
         29 . The composition of  claim 16 , wherein the allergen is a polypeptide which is secreted into a periplasm or secreted outside the cell.  
     
     
         30 . The composition of  claim 16 , wherein the microorganism is a gram-negative bacteria or yeast that secretes the allergen into a periplasm.  
     
     
         31 . A pharmaceutical composition comprising microorganisms that produce an allergen that elicits an anaphylactic allergic response in a subject susceptible to the anaphylactic allergic response, and further comprises an pharmaceutically acceptible carrier.  
     
     
         32 . The pharmaceutical composition of  claim 31 , wherein the allergen is a polypeptide or a small molecule.  
     
     
         33 . The pharmaceutical composition of  claim 31 , wherein the microorganisms produce a portion of an allergen that elicits an anaphylactic allergic response in a subject susceptible to the anaphylactic allergic response.  
     
     
         34 . A composition comprising dead  E. coli  containing therein at least one modified peanut allergen whose amino acid sequence differs from that of a wild-type peanut allergen that occurs in nature such that the modified peanut allergen has a reduced ability to bind to or cross-link IgE as compared with the wild-type peanut allergen.  
     
     
         35 . The composition of  claim 34 , wherein the wild-type peanut allergen is Ara h 1 (SEQ ID NO:1).  
     
     
         36 . The composition of  claim 34 , wherein the wild-type peanut allergen is Ara h 2 (SEQ ID NO:2).  
     
     
         37 . The composition of  claim 34 , wherein the wild-type peanut allergen is Ara h 3 (SEQ ID NO:3).  
     
     
         38 . The composition of  claim 34 , wherein the sequence of the modified peanut allergen differs from the sequence of the wild-type peanut allergen by one or more amino acid deletions, substitutions or additions within an IgE binding site of the wild-type peanut allergen.  
     
     
         39 . The composition of  claim 38 , wherein the sequence of the modified peanut allergen lacks a portion of the wild-type peanut allergen sequence, and wherein said portion includes an IgE binding site.  
     
     
         40 . The composition of  claim 34 , wherein the modified peanut allergen is located in the cytoplasm of the dead  E. coli.    
     
     
         41 . The composition of  claim 34 , wherein the modified peanut allergen is located in periplasm of the dead  E. coli.    
     
     
         42 . The composition of  claim 34 , wherein the modified peanut allergen cannot be by antibody binding without disrupting the dead  E. coli.