Methods and compositions for screening for modulators of IgE synthesis, secretion and switch rearrangement

Abstract

The invention relates to methods and compositions useful in screening for modulators of IgE synthesis, secretion and switch rearrangement.

1 - 28 . (canceled)  
     
     
         29 . A cultured mammalian cell comprising: 
 a nuclear genome comprising a recombinant nucleic acid comprising: 
 a) an IL-4 inducible fragment of an IgG epsilon promoter; and  
 b) a polynucleotide encoding a reporter protein;  
   wherein said IL-4 inducible fragment of said IgG epsilon promoter and said polynucleotide encoding said reporter protein are operably linked such that expression of said reporter protein is IL-4 inducible.    
     
     
         30 . The cultured mammalian cell of  claim 29 , wherein said cultured mammalian cell is an IgM +  B cell.  
     
     
         31 . The cultured mammalian cell of  claim 29 , wherein said reporter protein is an optically detectable protein, a death protein, or a survival protein.  
     
     
         32 . The cultured mammalian cell of  claim 29 , wherein said reporter protein is an optically detectable protein.  
     
     
         33 . The cultured mammalian cell of  claim 29 , wherein said reporter protein is a fusion protein.  
     
     
         34 . The cultured mammalian cell of  claim 33 , wherein said fusion protein comprises two reporter domains.  
     
     
         35 . The cultured mammalian cell of  claim 33 , wherein said fusion protein comprises an IgE heavy chain domain and reporter domain.  
     
     
         36 . The cultured mammalian cell of  claim 33 , wherein said IgG epsilon promoter is endogenous to said nuclear genome.  
     
     
         37 . The cultured mammalian cell of  claim 33 , wherein said IgG epsilon promoter is exogenous to said nuclear genome.  
     
     
         38 . The cultured mammalian cell of  claim 29 , further comprising nucleic acid encoding a test peptide.  
     
     
         39 . The cultured mammalian cell of  claim 29 , wherein said peptide comprises a random sequence of amino acids.  
     
     
         40 . A method of using the cultured mammalian cell of  claim 29 , comprising: 
 contacting said cultured mammalian cell with a test agent; and    detecting an effect of said test agent on IL-4 inducible expression of said reporter protein.    
     
     
         41 . The method of  claim 40 , wherein said agent is an organic molecule having a molecular weight of less than 2500 Da.  
     
     
         42 . The method of  claim 40 , wherein said contacting comprises introducing said test agent into said cell.  
     
     
         43 . The method of  claim 40 , wherein said agent is a polypeptide or polynucleotide page 19, lines 23-25  
     
     
         44 . The method of  claim 40 , wherein said nucleic acid encodes a random peptide.  
     
     
         45 . A method of making the cultured mammalian cell of  claim 29 , comprising: 
 introducing into a cell a vector comprising a recombinant nucleic acid comprising: 
 a) an IL-4 inducible fragment of an IgG epsilon promoter; and  
 b) a polynucleotide encoding a reporter protein; and  
   culturing said cell under conditions for selection for cells in which said recombinant nucleic acid has integrated into said genome.    
     
     
         46 . The method of  claim 45 , further comprising selecting a cell that does not detectably produce said reporter protein in the absence of IL-4.  
     
     
         47 . The method of  claim 45 , wherein said vector is a retroviral vector.