Localization of major peptide autoepitopes for nucleosome specific T cells of systemic lupus erythematosus

Abstract

The present invention includes peptides derived from nucleosomal histone proteins which are useful for delaying the onset and progression of systemic lupus erythematosus (i.e. lupus or SLE). The peptides of the invention span the histone proteins (i.e. H1, H2A, H2B, H3, and H4). The invention additionally encompasses isolated nucleic acids which encode these histone peptides as well as pharmaceutical compositions which comprise one or more of a histone peptide. Further, the invention provides kits which comprise one or more histone peptides or isolated nucleic acids encoding histone peptides and an instructional material. The invention also provides methods of using these compositions and analogs of histone peptides to inhibit an immune response and associated inflammation in an animal and to treat disorders in an animal which are related to the production of autoantibodies and complications thereof, such as inflammatory diseases, autoimmune disorders, and nephritis. In addition, the invention includes methods for developing diagnostic and prognostic reagents using the histone peptides and isolated nucleic acids encoding the instone peptides, for the purpose of tracking autoimmune T helper cells and B cells of SLE are also included.

What is claimed is:  
     
         1 . A composition comprising an isolated peptide wherein said peptide has an amino acid sequence corresponding to the amino acid sequence of a portion of a nucleosome histone protein and, wherein said isolated peptide is capable of promoting immunological tolerance in an animal having systemic lupus erythematosus.  
     
     
         2 . The composition of  claim 1 , wherein said nucleosome histone protein is selected from the group consisting of histone 1 (H1), histone 2A (H2A), histone 2B (H2B), histone 3 (H3), and histone 4 (H4).  
     
     
         3 . The composition of  claim 1 , said peptide comprising not more than 27 contiguous amino acids and having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, and 26.  
     
     
         4 . A composition comprising the isolated peptide of  claim 1  and a pharmaceutically acceptable carrier.  
     
     
         5 . The composition of  claim 1 , wherein said portion of said histone protein corresponds to an autoepitope, which autoepitope is associated with systemic lupus erythematosus and recognized by one or more of an autoimmune T cell and an autoimmune B cell.  
     
     
         6 . An isolated nucleic acid encoding the peptide of  claim 1 .  
     
     
         7 . The isolated nucleic acid of  claim 6 , said nucleic acid comprising a nucleotide sequence selected from the group consisting of SEQ ID NOS.: 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51 and 52.  
     
     
         8 . A cell comprising the isolated nucleic acid of  claim 6 .  
     
     
         9 . The cell of  claim 8 , wherein said cell is selected from the group consisting of a prokaryotic cell and a eukaryotic cell.  
     
     
         10 . The cell of  claim 9 , wherein said cell is an insect cell.  
     
     
         11 . A vector comprising the isolated nucleic acid of  claim 6 .  
     
     
         12 . The composition of  claim 1 , wherein said peptide further comprises a covalently attached moiety selected from the group consisting of a fluorophore, a chromophore, a biotin moiety, a light reactive group, and an enzyme cleavable group.  
     
     
         13 . A method of treating an animal having an autoimmune disorder, said method comprising administering to said animal an isolated peptide comprising a portion of a nucleosome histone protein, wherein said isolated peptide is capable of promoting immunological tolerance in an animal, thereby treating said autoimmune disorder.  
     
     
         14 . The method of  claim 13 , wherein said autoimmune disorder is selected from the group consisting of rheumatoid arthritis, scleroderma, and systemic lupus erythematosus.  
     
     
         15 . The method of  claim 14 , wherein said autoimmune disorder is systemic lupus erythematosus.  
     
     
         16 . The method of  claim 13 , wherein said isolated peptide is administered in an amount which is from at least about 10 micrograms per kilogram of animal to at least about 1 gram per kilogram of animal.  
     
     
         17 . The method of  claim 16 , wherein said amount is from at least about 100 micrograms per kilogram of animal to about 600 micrograms per kilogram of animal.  
     
     
         18 . The method of  claim 13 , wherein said peptide comprises not more than 27 contiguous amino acids and having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, and 26.  
     
     
         19 . A method of treating nephritis in an animal having systemic lupus erythematosus, said method comprising administering to said animal an isolated peptide comprising a portion of a nucleosome histone protein, wherein said isolated peptide is capable of promoting immunological tolerance in an animal, thereby alleviating said nephritis in said animal.  
     
     
         20 . A method of reducing the production of autoantibodies in an animal, said method comprising administering to said animal an isolated peptide comprising a portion of a nucleosome histone protein, wherein said isolated peptide is capable of promoting immunological tolerance in an animal, and wherein said peptide is administered in an amount sufficient to promote immunologic tolerance in said animal, thereby reducing the production of autoantibodies in said animal.  
     
     
         21 . A method of treating inflammation in an animal, which inflammation is caused by the production of autoantibodies in said animal, said method comprising administering to said animal an isolated peptide comprising a portion of a nucleosome histone protein, wherein said isolated peptide is capable of promoting immunological tolerance in an animal, and wherein said peptide is administered in an amount sufficient to promote immunological tolerance in said animal, thereby inhibiting the production of autoantibodies in said animal and alleviating inflammation in said animal.  
     
     
         22 . A method of diagnosing systemic lupus erythematosus in an animal, said method comprising 
 (a) contacting a sample from said animal with a composition comprising an isolated histone peptide complex, wherein said histone peptide complex comprises 
 i) a histone peptide portion, histone peptide portion comprising no more than 27 contiguous amino acids and having an amino acid sequence corresponding to a portion of a nucleosome histone protein;  
 ii) a fused portion having an amino acid sequence which corresponds to a portion of a protein selected from the group consisting of a major histocompatibility class II molecule and an immunoglobin; and  
 iii) an indicator portion, wherein said indicator portion is a molecule which is capable of producing a detectable chemical signal, and which is selected from the group consisting of a flourophore, a chromophore, a light reactive moiety and a biotin moiety; and wherein each of said histone peptide portion, said fused portion, and said indication portion is covalently linked to at least one other component of the histone peptide complex; and  
   (b) identifying in said animal said signal produced by said indicator portion, whereby the identification of said signal in said animal is an indication that said animal has systemic lupus erythematosus, thereby diagnosing systemic lupus erythematosus in said animal.    
     
     
         23 . A method of tracking an autoimmune cell associated with systemic lupus erythematosus in an animal, said method comprising 
 (a) contacting a sample from said animal with a composition comprising one or more of a modified histone peptide and an isolated histone peptide complex, wherein said modified histone peptide comprises 
 (i) an modified portion wherein said modified portion is a molecule which is capable of producing a detectable chemical signal, and which is selected from the group consisting of a flourophore, a chromophore, a light reactive moiety and a biotin moiety; and  
 (ii) a peptide portion comprising not more than 27 contiguous amino acids and having an amino acid sequence corresponding to a nucleosome histone protein,  
 wherein said peptide portion and said modified portion are covalently linked; and  
 wherein said histone peptide complex comprises  
   (iii) a histone peptide portion, histone peptide portion comprising no more than 27 contiguous amino acids and having an amino acid sequence corresponding to a portion of a nucleosome histone protein; 
 (iv) a fused portion having an amino acid sequence which corresponds to a portion of a protein selected from the group consisting of a major histocompatibility class II molecule and an immunoglobin; and  
 (v) an indicator portion, wherein said indicator portion is a molecule which is capable of producing a detectable chemical signal, and which is selected from the group consisting of a flourophore, a chromophore, a light reactive moiety and a biotin moiety; and  
 wherein each of said histone peptide portion, said fused portion, and said indicator portion is covalently linked to at least one other component of the histone peptide complex; and  
   (b) identifying and monitoring in said animal said signal produced by said indicator portion, wherein said identification and monitoring of said signal in said animal constitutes the identification and monitoring of an autoimmune cell associated with systemic lupus erythematosus to which said histone protein complex is bound, thereby tracking said autoimmune cell associated with systemic lupus erythematosus in said animal.    
     
     
         24 . A method of treating an animal having an autoimmune disorder, said method comprising administering to said animal a modified histone peptide, wherein said modified histone peptide is an altered peptide ligand, and wherein said modified histone peptide is capable of promoting immunological tolerance in an animal, thereby treating said autoimmune disorder.  
     
     
         25 . The method of  claim 24 , wherein said autoimmune disorder is selected from the group consisting of rheumatoid arthritis, scleroderma, and systemic lupus erythematosus.  
     
     
         26 . The method of  claim 25 , wherein said autoimmune disorder is systemic lupus erythematosus.  
     
     
         27 . The method of  claim 24 , wherein said isolated peptide is administered in an amount which is from at least about 10 micrograms per kilogram of animal to at least about 1 gram per kilogram of animal.  
     
     
         28 . The method of  claim 27 , wherein said amount is from at least about 100 micrograms per kilogram of animal to about 600 micrograms per kilogram of animal.  
     
     
         29 . The method of  claim 24 , wherein said modified peptide comprises not more than 27 contiguous amino acids and has an amino acid sequence which is at least one amino acid different relative to an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, and 26.  
     
     
         30 . A method of treating nephritis in an animal having systemic lupus erythematosus, said method comprising administering to said animal a modified histone peptide, wherein said modified histone peptide is an altered peptide ligand, and wherein said modified histone peptide is capable of promoting immunological tolerance in an animal, thereby alleviating said nephritis in said animal.  
     
     
         31 . A method of reducing the production of autoantibodies in an animal, said method comprising administering to said animal a modified histone peptide, wherein said modified histone peptide is an altered peptide ligand capable of promoting immunological tolerance in an animal, and wherein said modified histone peptide is administered in an amount sufficient to promote immunologic tolerance in said animal, thereby reducing the production of autoantibodies in said animal.  
     
     
         32 . A method of treating inflammation in an animal, which inflammation is caused by the production of autoantibodies in said animal, said method comprising administering to said animal a modified histone peptide, wherein said modified histone peptide is an altered peptide ligand capable of promoting immunological tolerance in an animal, and wherein said modified histone peptide is administered in an amount sufficient to promote immunological tolerance in said animal, thereby inhibiting the production of autoantibodies in said animal and alleviating inflammation in said animal.