Mrgprx2/mrgprb2 expressing cell based assay to detect pseudo-allergic drug reactions and to identify blockers to prevent the adverse reactions

Abstract

The present invention relates to cells and methods for detecting compounds that induce a pseudo-allergic-type reaction and methods for reducing the severity of a pseudo-allergic-type reaction.

1 - 22 . (canceled) 
     
     
         23 . A method for identifying an antagonist of MrgprX2 or MrgprB2 comprising:
 contacting an isolated cell comprising a recombinant nucleic acid that expresses mas-related G-protein coupled receptor member X2 (MrgprX2) or MrgprB2 with a compound that induces a pseudo-allergic-type reaction,   contacting the isolated cell with a candidate antagonist,   detecting activation of MrgprX2 or MrgprB2, wherein a decrease in activation of MrgprX2 or MrgprB2 relative to the activation of MrgprX2 or MrgprB2 in the absence of the compound determines that the candidate compound is an antagonist.   
     
     
         24 . The method of  claim 23 , wherein the recombinant nucleic acid expresses MrgprX2. 
     
     
         25 . The method of  claim 23 , wherein the recombinant nucleic acid expresses MrgprB2. 
     
     
         26 . The method of  claim 24 , wherein the recombinant nucleic acid that expresses MrgprX2 comprises one or more mutations. 
     
     
         27 . The method of  claim 26 , wherein the one or more mutations produces an MrgprX2 protein incapable of activating a signal transduction pathway. 
     
     
         28 . The method of  claim 25 , wherein the recombinant nucleic acid that expresses MrgprB2 comprises one or more mutations. 
     
     
         29 . The method of  claim 28 , wherein the one or more mutations produces an MrgprB2 protein incapable of activating a signal transduction pathway. 
     
     
         30 . The method of  claim 23 , wherein the isolated cell comprises a human embryonic kidney 293 (HEK 293) cell. 
     
     
         31 . The method of  claim 23  wherein the MrgprB2 comprises the amino acid sequence of SEQ ID NO: 3. 
     
     
         32 . The method of  claim 23  wherein the MrgprX2 comprises the amino acid sequence of SEQ ID NO: 1.