Methods of screening for modulators of HIV infection

Abstract

The present invention provides polynucleotides that encode the chemokine receptors 88-2B or 88C and materials and methods for the recombinant production of these two chemokine receptors. Also provided are assays utilizing the polynucleotides which facilitate the identification of ligands and modulators of the chemokine receptors. Receptor fragments, ligands, modulators, and antibodies are useful in the detection and treatment of disease states associated with the chemokine receptors such as atherosclerosis, rheumatoid arthritis, tumor growth suppression, asthma, viral infection, AIDS, and other inflammatory conditions.

We claim:  
     
         1 . A method of screening for a modulator of human immunodeficiency virus (HIV) or simian immunodeficiency virus (SIV) infection, comprising steps of: 
 (a) contacting a first composition comprising an 88C receptor polypeptide with a second composition comprising a HIV or SIV envelope protein in the presence and absence of a compound, wherein the 88C receptor polypeptide comprises an amino acid sequence encoded by a nucleotide sequence that hybridizes to the complement of nucleotides 55-1110 of SEQ ID NO: 1 or the complement of nucleotides 1-1056 of SEQ ID NO: 19 under the following stringent conditions: 42° C. in 50% formamide, 5X SSC, 20 mM sodium phosphate, pH 6.8 and washing in 0.2X SSC at 55° C.;    (b) measuring the interaction of the 88C receptor polypeptide with the envelope protein in the presence and absence of the compound; and    (c) screening for a modulator of HIV or SIV infection, wherein reduced or increased interaction of the 88C receptor polypeptide with the envelope protein in the presence of the compound versus in the absence of the compound is indicative of the compound being a modulator of HIV or SIV infection.    
     
     
         2 . A method according to  claim 1 , wherein the 88C receptor polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 2.  
     
     
         3 . A method according to  claim 2  wherein the envelope protein comprises an HIV envelope protein.  
     
     
         4 . A method according to  claim 1 , wherein the 88C receptor polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 20.  
     
     
         5 . A method according to  claim 4 , wherein the envelope protein comprises an SIV envelope protein.  
     
     
         6 . A method according to  claim 1 , wherein the first composition comprises a cell that has been recombinantly modified to express the 88C receptor polypeptide on its surface.  
     
     
         7 . A method according to  claim 6 , wherein the second composition comprises HIV which comprises HIV envelope protein.  
     
     
         8 . A method according to  claim 7 , wherein the measuring step comprises measuring HIV infection of the cell.  
     
     
         9 . A method according to  claim 1 , further comprising a step of forming a modulator composition by mixing a modulator identified according to step (c) with a pharmaceutically acceptable diluent or carrier.  
     
     
         10 . A method according to  claim 9 , further comprising a step of administering the modulator composition to a mammalian subject, and screening for modulation of HIV or SIV infection in said subject.  
     
     
         11 . A method of screening for a modulator of human immunodeficiency virus (HIV) infection, comprising steps of: 
 (a) contacting a first composition comprising an 88-2B receptor polypeptide with a second composition comprising a HIV envelope protein, in the presence and absence of a compound, wherein the 88-2B receptor polypeptide comprises an amino acid sequence encoded by a nucleotide sequence that hybridizes to the complement of nucleotides 362-1426 of SEQ ID NO:3 under the following stringent conditions: 42° C. in 50% formamide, 5X SSC, 20 mM sodium phosphate, pH 6.8 and washing in 0.2X SSC at 55° C.;    (b) measuring the interaction of the 88-2B receptor polypeptide with the HIV envelope protein in the presence and absence of the compound; and    (c) screening for a modulator of HIV infection, wherein reduced or increased interaction of the 88-2B receptor polypeptide with the HIV envelope protein in the presence of the compound versus in the absence of the compound is indicative of the compound being a modulator of HIV infection.    
     
     
         12 . A method according to  claim 11 , wherein the 88-2B receptor polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 4.  
     
     
         13 . A method according to  claim 12 , wherein the first composition comprises a cell that has been recombinantly modified to express the 88-2B receptor polypeptide on its surface.  
     
     
         14 . A method according to  claim 13 , wherein the second composition comprises a human immunodeficiency virus which comprises HIV envelope protein.  
     
     
         15 . A method according to  claim 14 , wherein the measuring step comprises measuring HIV infection of the cell.  
     
     
         16 . A method according to  claim 12 , further comprising a step of forming a modulator composition by mixing a modulator identified according to step (c) with a pharmaceutically acceptable diluent or carrier.  
     
     
         17 . A method according to  claim 16 , further comprising a step of administering the modulator composition to a mammalian subject, and screening for modulation of HIV infection in said subject.  
     
     
         18 . A method for detecting human immunodeficiency virus (HIV) infection of cells, comprising steps of: 
 (a) contacting a cell that has been recombinantly modified to express at least one of human chemokine receptors 88C and 88-2B with HIV; and    (b) detecting HIV infection of the cell.    
     
     
         19 . A method for inhibiting human immunodeficiency virus (HIV) infection of cells, comprising steps of: 
 (a) contacting cells with an antibody to at least one of human chemokine receptors 88C and 88-2B with HIV; and    (b) detecting HIV infection of the cell after said contacting step.