Method to regulate CD40 signaling

Abstract

The present invention relates to methods useful for identifying compounds capable of specifically controlling CD40 regulation of JNK or p38 activity useful for inhibiting immunoglobulin heavy chain class switching, cytokine production and activation of cells involved in an inflammatory response. The present invention also includes kits to perform such assays and methods to control disease related to such responses.

What is claimed is:  
     
         1 . A method to identify a compound that controls CD40 regulation of JNK activity in a cell, comprising: 
 (a) contacting a cell with a putative regulatory compound, wherein the cell includes a CD40 protein and a Jun kinase protein; and    (b) assessing the ability of said putative regulatory compound to regulate the activity of said Jun kinase.    
     
     
         2 . The method of    claim 1   , wherein said assessment step comprises determining the ability of said JUN kinase protein to regulate the activity of a protein selected from the group consisting of c-Jun, ATF-2, Ets-1 and other Ets family members activity in said cell.  
     
     
         3 . The method of    claim 1   , wherein said method further comprises stimulating said cell, prior to said assessing step, with a ligand of CD40.  
     
     
         4 . The method of    claim 1   , wherein said method further comprises stimulating said cell, prior to said assessment step, with a stimulatory compound selected from the group consisting of soluble gp39, membrane-bound gp39 and an antibody that binds specifically to CD40.  
     
     
         5 . The method of    claim 1   , wherein said cell is selected from the group consisting of a mammalian, an invertebrate, a plant, an insect, a fungal, a yeast and a bacterial cell.  
     
     
         6 . The method of    claim 1   , wherein said cell is selected from the group consisting of a mammalian, an amphibian and a yeast cell.  
     
     
         7 . The method of    claim 1   , wherein said cell is selected from the group consisting of a primate, a mouse and a rat cell.  
     
     
         8 . The method of    claim 1   , wherein said cell is selected from the group consisting of Pre-B cells, B lymphocytes, cancer cells, fibroblasts, Langerhans cells, epithelial cells monocytes and dendritic cells.  
     
     
         9 . The method of    claim 1   , wherein said putative regulatory compound is selected from the group consisting of a protein-based compound, a carbohydrate-based compound, a lipid-based compound, a nucleic acid-based compound, a natural organic compound, a synthetically derived organic compound, an anti-idiotypic antibody and/or catalytic antibody, or fragments thereof.  
     
     
         10 . The method of    claim 1   , wherein said putative regulatory compound is selected from the group consisting of a small organic molecule, a peptide and a polypeptide.  
     
     
         11 . A method to identify a compound that controls CD40 regulation of p38 activity in a cell, comprising: 
 (a) contacting a cell with a putative regulatory compound, wherein the cell includes CD40 protein and p38 protein; and    (b) assessing the ability of said putative regulatory compound to regulate the activity of said p38 protein.    
     
     
         12 . The method of    claim 11   , wherein said assessment step comprises determining the ability of said p38 protein to regulate the activity of a protein selected from the group consisting of ATF-2, Ets-1 and other Ets family members activity in said cell.  
     
     
         13 . The method of    claim 11   , wherein said method further comprises stimulating said cell, prior to said assessing step, with a ligand of CD40.  
     
     
         14 . The method of    claim 11   , wherein said method further comprises stimulating said cell, prior to said assessment step, with a stimulatory compound selected from the group consisting of soluble gp39, membrane-bound gp39 and an antibody that binds specifically to CD40.  
     
     
         15 . The method of    claim 11   , wherein said cell is selected from the group consisting of Pre-B cells, B lymphocytes, cancer cells, fibroblasts, Langerhans cells, epithelial cells monocytes and dendritic cells.  
     
     
         16 . The method of    claim 11   , wherein said putative regulatory compound is selected from the group consisting of a small organic molecule, a peptide and a polypeptide.  
     
     
         17 . A regulatory compound identified by said compound's ability to regulate a biological function selected from the group consisting of immunoglobulin heavy chain class switching, cytokine production and inflammatory cell activation, said compound being capable of penetrating the plasma membrane of a cell and of inhibiting the ability of CD40 protein to regulate JNK protein activity in said cell.  
     
     
         18 . The compound of    claim 17   , wherein said compound is capable of regulating the activity of a protein selected from the group consisting of CD40 and Jun kinase protein.  
     
     
         19 . The compound of    claim 17   , wherein said regulation involves a step selected from the group consisting of sequestering JNK protein in an inactive complex, regulating the ligand binding activity of CD40, regulating MEKK activity, regulating the phosphorylation of JNK protein, regulating the interaction between JNK and JNKK, regulating the ability of JNK to activate c-Jun, ATF-2, and Ets-1 and other Ets family members, regulating the, expression of endogenous and/or heterologous nucleic acid molecules encoding CD40 or JNK protein, and combinations thereof.  
     
     
         20 . A regulatory compound capable of penetrating the plasma membrane of a cell and of inhibiting the ability of CD40 protein to regulate p38 protein activity in said cell, said compound able to regulate a biological function selected from the group consisting of immunoglobulin heavy chain class switching, cytokine production and inflammatory cell activation.  
     
     
         21 . The compound of    claim 20   , wherein said compound is capable of regulating the activity of a protein selected from the group consisting of CD40 and p38 protein.  
     
     
         22 . The compound of    claim 20   , wherein said regulation involves a step selected from the group consisting of sequestering p38 protein in an inactive complex, regulating the ligand binding activity of CD40, regulating MEKK activity, regulating the phosphorylation of p38 protein, regulating the interaction between p38 and MEK, regulating the ability of p38 to activate ATF-2, and Ets-1 and other Ets family members, regulating the expression of endogenous and/or heterologous nucleic acid molecules encoding CD40 or p38 protein, and combinations thereof.  
     
     
         23 . A method to inhibit immunoglobulin heavy chain class switching, comprising inhibiting the activity of a protein selected from the group consisting of Jun kinase protein and p38 protein.  
     
     
         24 . The method of    claim 23   , wherein said activity being inhibited is selected from the group consisting of sequestering JNK or p38 protein in an inactive complex, regulating the ligand binding activity of CD40, regulating MEKK activity, regulating the phosphorylation of JNK or p38 protein, regulating the interaction between JNK and JNKK, regulating the ability of JNK to activate c-Jun, ATF-2, and Ets-1 and other Ets family members, regulating the interaction between p38 and MEK, regulating the ability of p38 to activate ATF-2, and Ets-1 and other Ets family members, regulating the expression of endogenous or heterologous nucleic acid molecules encoding a CD40, JNK or p38 protein, and combinations thereof.  
     
     
         25 . The method of    claim 24   , wherein said activity is inhibited using a compound selected from the group consisting of a Jun kinase activation site mimetope, a Jun kinase target site mimetope, a Jun kinase kinase domain mimetope, a mutated Jun kinase protein, a Jun kinase pseudosubstrate, a p38 activation site mimetope, a p38 target site mimetope, a p38 kinase domain mimetope, a mutated p38 protein, a p38 pseudosubstrate, a mutated CD40 protein, a CD40 antagonist, an antisense oligonucleotide, a ribozyme and an expression plasmid encoding CD40 or Jun kinase.  
     
     
         26 . A method to inhibit cytokine production by a cell having CD40, comprising inhibiting the activity of a protein selected from the group consisting of Jun kinase protein and p38 protein.  
     
     
         27 . The method of    claim 26   , wherein said activity is inhibited using a compound selected from the group consisting of a Jun kinase activation site mimetope, a Jun kinase target site mimetope, a Jun kinase kinase domain mimetope, a mutated Jun kinase protein, a Jun kinase pseudosubstrate, a p38 activation site mimetope, a p38 target site mimetope, a p38 kinase domain mimetope, a mutated p38 protein, a p38 pseudosubstrate, a mutated CD40 protein, a CD40 antagonist, an antisense oligonucleotide, a ribozyme and an expression plasmid encoding CD40 or Jun kinase.  
     
     
         28 . A method to treat an animal with a disease selected from the group consisting of a disease involving an allergic response and a disease involving an autoimmune disease, said method comprising administering to an animal an effective amount of a therapeutic composition comprising a compound that controls CD40 regulation of the activity of a protein selected from the group consisting of Jun kinase and p38 protein.  
     
     
         29 . The method of    claim 28   , wherein said disease involving an allergic response is selected from the group consisting of asthma, allergic rhinitis, allergic hypersensitivity, atopic dermatitis and acute bronchopulmonary aspergillosis.  
     
     
         30 . The method of    claim 28   , wherein said disease involving an autoimmune disease is selected from the group consisting of rheumatoid arthritis and systemic lupus erythematosus.  
     
     
         31 . A kit to identify compounds that controls CD40 regulation of JNK activity in a cell, said kit comprising: 
 (a) a cell comprising a CD40 protein and a Jun kinase protein; and    (b) a means for detecting regulation of said Jun kinase protein.    
     
     
         32 . The kit of    claim 32   , wherein said cell is selected from the group consisting of Pre-B cells, B lymphocytes, cancer cells, fibroblasts, Langerhans cells, epithelial cells monocytes and dendritic cells.  
     
     
         33 . A kit to identify compounds that controls CD40 regulation of p38 activity in a cell, said kit comprising: 
 (a) a cell comprising CD40 protein and p38 protein; and    (b) a means for detecting regulation of said p38 protein.    
     
     
         34 . The kit of    claim 33   , wherein said cell is selected from the group consisting of Pre-B cells, B lymphocytes, cancer cells, fibroblasts, Langerhans cells, epithelial cells monocytes and dendritic cells.