Nef-attachable protein, DNA encoding the protein and a monoclonal antibody against said protein

Abstract

A monoclonal antibody specific to Nap was prepared by using a crude membrane fraction from a cell line which has a high affinity to Nef as an antigen. A cDNA clone which encodes Nap was obtained by screening a cDNA library of said cell line utilizing the specific antibody. The nucleotide sequence of this cDNA was elucidated whereupon the full amino acid sequences of human Nap was identified. Since the anti-Nap monoclonal antibody inhibits the binding of Nef to Nap, it can be used as a new therapeutic agent for AIDS. In addition, it can be applied for the diagnosis of AIDS by clarifying the relation between the expression of Nap and the development of AIDS. Nap can be utilized for binding experiments to Nef. It is useful for the development of novel therapeutic agents for AIDS based upon a mechanism of action which inhibits the attachment of both factors and development of new inhibitors against the onset of AIDS by a mechanism of action different from that of reverse transcriptase inhibitors or protease inhibitors.

1 - 8 . (Canceled).  
     
     
         9 . A monoclonal antibody against a protein having an amino acid sequence represented by SEQ ID NO:1 or a protein having an amino acid sequence represented by SEQ ID NO:1, wherein one amino acid residue is deleted, added, or substituted, and the protein binds to Nef.  
     
     
         10 . A monoclonal antibody as claimed in  claim 9  against a protein having an amino acid sequence represented by SEQ ID NO:1, wherein one amino acid residue is deleted, added, or substituted, and the protein binds to Nef.  
     
     
         11 . A monoclonal antibody as claimed in  claim 9  which is a mouse monoclonal antibody.  
     
     
         12 . A chimeric antibody having a variable region of the mouse monoclonal antibody according to  claim 11  and a constant region of a human type antibody.  
     
     
         13 . A pharmaceutical agent containing an antibody according to  claim 12  as an effective ingredient.  
     
     
         14 . A pharmaceutical agent according to  claim 13  where said agent is a therapeutic agent against AIDS.  
     
     
         15 . A human type antibody having a complementary-determining region of the mouse monoclonal antibody according to  claim 11 .  
     
     
         16 . A pharmaceutical agent containing an antibody according to  claim 15  as an effective ingredient.  
     
     
         17 . A pharmaceutical agent according to  claim 16  where said agent is a therapeutic agent against AIDS.  
     
     
         18 . An anti-Nef-attachable protein monoclonal antibody which has a high affinity to Molt-4 clone no.8 cells (human CD4 +  T cell line) and U937 cells (human macrophage cell line) and does not bind or attach to Raji cells (human B cell line), BT-2 cells (human gliocyte cell line) and Gin-1 cells (human fibroblast cell line).  
     
     
         19 . A monoclonal antibody as claimed in  claim 18  which is a mouse monoclonal antibody.  
     
     
         20 . A chimeric antibody having a variable region of the mouse monoclonal antibody according to  claim 19  and a constant region of a human type antibody.  
     
     
         21 . A human type antibody having a complementary-determining region of the mouse monoclonal antibody according to  claim 19 .  
     
     
         22 . A pharmaceutical agent containing an antibody according to  claim 20  as an effective ingredient.  
     
     
         23 . A pharmaceutical agent according to  claim 22  where said agent is a therapeutic agent against AIDS.  
     
     
         24 . A pharmaceutical agent containing an antibody according to  claim 21  as an effective ingredient.  
     
     
         25 . A pharmaceutical agent according to  claim 24  where said agent is a therapeutic agent against AIDS.  
     
     
         26 . A method to diagnose the development of AIDS comprising using a protein having an amino acid sequence represented by SEQ ID NO:1 or a protein having an amino acid sequence represented by SEQ ID NO:1 wherein one amino acid residue is deleted, added, or substituted, and the protein binds to Nef as an index.  
     
     
         27 . A method to diagnose the development of AIDS comprising using a DNA molecule which encodes a protein represented by SEQ ID NO:1 or which encodes a protein having an amino acid sequence where one amino acid residue in the amino acid sequence represented by SEQ ID NO:1 is deleted, added or substituted, and the protein binds to Nef.  
     
     
         28 . A method for screening for a therapeutic agent against AIDS comprising using a Nef-binding property of a protein having an amino acid sequence represented by SEQ ID NO:1 or a protein having an amino acid sequence represented by SEQ ID NO:1, wherein one amino acid residue is deleted, added, or substituted, and the protein binds to Nef.  
     
     
         29 . A monoclonal antibody as claimed in  claim 9  which is against a protein having an amino acid sequence represented by SEQ. ID NO:1.  
     
     
         30 . A monoclonal antibody against a recombinant protein having an amino acid sequence represented by SEQ ID NO:1 or against a recombinant protein having an amino acid sequence represented by SEQ ID NO:1, wherein one amino acid residue is deleted, added, or substituted, and the protein binds to Nef.