Multimeric forms of human rhinovirus receptor protein

Abstract

The present invention relates to novel forms and configurations of intercellular adhesion molecule (ICAM) including multimeric configurations that effectively bind to human rhinovirus and can effectively reduce HRV infectivity. When in a multimeric configuration, preferably as dimers, these proteins display enhanced binding of HRV and are able to reduce HRV infectivity as well as the infectivity of other viruses known to bind to the “major” group human rhinovirus receptor (HRR). The multimerized proteins may also be used to block tICAM interaction with lymphocyte function-associated antigen-1 (LFA-1).

What is claimed is:  
     
         1 . Multimeric ICAM.  
     
     
         2 . The multimeric ICAM of  claim 1  wherein said ICAM is non-transmembrane ICAM.  
     
     
         3 . The multimeric ICAM of  claim 2  wherein said non-transmembrane ICAM is substantially without the carboxyl intracellular domain and without the hydrophobic membrane domain.  
     
     
         4 . The multimeric ICAM according to  claim 2  wherein said non-transmembrane ICAM is a member selected from the group consisting of tICAM(453), tICAM(185), tICAM(88), tICAM(283), and tICAMs comprising one or more sequences selected from tICAM(89-185), tICAM186-283, tICAM(284-385), tICAM(386-453), tICAM(75-77), tICAM(70-72), tICAM(64-66), tICAM(40-43), tICAM(36-38), tICAM(30-33), and tICAM(26-29).  
     
     
         5 . The multimeric ICAM of  claim 1  wherein said ICAM is multimerized by adsorption to a support.  
     
     
         6 . The multimeric ICAM of  claim 5  wherein said support is an inert polymer and is a member selected from the group consisting of nitrocellulose, PVDF, DEAE, lipid polymer, and amino dextran.  
     
     
         7 . The multimeric ICAM of  claim 1  wherein said multimeric ICAM is multimerized by coupling to a member.  
     
     
         8 . The multimeric ICAM of  claim 7  wherein said ICAM is modified with at least one reactive amino acid to provide at least one site to facilitate coupling.  
     
     
         9 . The multimeric ICAM of  claim 8  wherein said reactive amino acid is a member selected from the group consisting of lysine and cysteine.  
     
     
         10 . The multimeric ICAM of  claim 7  wherein said member is a member selected from the group consisting of an antibody and a protein carrier.  
     
     
         11 . The multimeric ICAM of  claim 10  wherein said antibody is anti-ICAM antibody CL 203.  
     
     
         12 . The multimeric ICAM of  claim 10  wherein said protein carrier is a member selected from the group consisting of albumin and proteoglycans.  
     
     
         13 . The multimeric ICAM of  claim 1  wherein said ICAM is modified at either terminus to comprise a lipid capable of promoting formation of oligomer micelles.  
     
     
         14 . The multimeric ICAM of  claim 1  comprising two or more ICAMs, which may be the same or different, linked to each other.  
     
     
         15 . The multimeric ICAM of  claim 14  wherein said ICAMs are directly linked to each other without a linker.  
     
     
         16 . The multimeric ICAM of  claim 15  wherein said ICAMs are linked to each other via at least one disulfide bridge.  
     
     
         17 . The multimeric ICAM of  claim 16  wherein said ICAMs ate crosslinked via a cysteine disulfide bridge at position 307 on each ICAM.  
     
     
         18 . The multimeric ICAM of  claim 16  wherein said ICAMs are crosslinked via a cysteine disulfide bridge at position 309 on each ICAM.  
     
     
         19 . The multimeric ICAM of  claim 14  wherein said ICAMs are indirectly linked via a cross-linking agent.  
     
     
         20 . The multimeric ICAM of  claim 19  wherein said cross-linking agent is selected from the group consisting of heterobifunctional and homobifunctional cross-linking reagents.  
     
     
         21 . The multimeric ICAM of  claim 20  wherein said cross-linking reagent is a member selected from the group consisting of bifunctional N-hydroxysuccinimide esters, imidoesters and bis-maleimido-hexanes.  
     
     
         22 . The multimeric ICAM of  claim 1  wherein said ICAM is a member selected from the group consisting of fully glycosylated ICAM, partially glycosylated ICAM, or non-glycosylated ICAM.  
     
     
         23 . In a method for enhancing the binding of ICAM to a ligand, the improvement comprising the steps of: 
 presenting said ICAM in a multimeric configuration.    
     
     
         24 . The method according to  claim 23  wherein said ICAM is tICAM.  
     
     
         25 . The method according to  claim 24  wherein said ICAM is a member selected from the group consisting of tICAM(453), tICAM(185), tICAM(88), tICAM(283), and tICAMs comprising one or more sequences selected from tICAM(89-185), tICAM186-283, tICAM(284-385), tICAM(386-453), tICAM(75-77), tICAM(70-72), tICAM(64-66), tICAM(40-43), tICAM(6-38), tICAM(30-33), and tICAM(26-29).  
     
     
         26 . The method according to  claim 23  wherein said ICAM is modified with at least one reactive amino acid to provide at least one site to facilitate coupling.  
     
     
         27 . The method according to  claim 26  wherein said reactive amino acid is selected from the group consisting of lysine and cysteine.  
     
     
         28 . The method according to  claim 23  wherein said ICAM is modified at either terminus to comprise a lipid capable of promoting formation of oligomer micelles.  
     
     
         29 . The method according to  claim 23  wherein said multimeric configuration comprises a first ICAM cross-linked to a second ICAM.  
     
     
         30 . The method according to  claim 29  wherein said first and second ICAM are each muteinized to contain a cysteine residue at position 307, and said first and second ICAM are cross-linked via a disulfide bridge between said cysteines at position 307.  
     
     
         31 . The method according to  claim 29  wherein said first and second ICAM are each muteinized to contain a cysteine residue at position 309, and said first and second ICAM are cross-linked via a disulfide bridge between said cysteines at position 309.  
     
     
         32 . The method according to  claim 23  wherein said multimeric configuration comprises ICAM adsorbed to a support.  
     
     
         33 . The method according to  claim 32  wherein said support comprises a member selected from the group consisting of high molecular weight and substantially inert polymers.  
     
     
         34 . The method according to  claim 33  wherein said polymer is an inert polymer and is a member selected from the group consisting of nitrocellulose, PVDF, DEAE, lipid polymers, and amino dextran.  
     
     
         35 . The method according to  claim 33  wherein said multimeric ICAM is multimerized by coupling to a member.  
     
     
         36 . The method according to  claim 35  wherein said member is a member selected from the group consisting of an antibody and a protein carrier.  
     
     
         37 . The method according to  claim 29  wherein said cross-linking reagent is a member selected from the group consisting of heterobifunctional and homobifunctional cross-linking reagents.  
     
     
         38 . The method according to  claim 37  wherein said protein carrier is a member selected from the group consisting of albumin and proteoglycans.  
     
     
         39 . The method according to  claim 36  wherein said antibody is anti-ICAM antibody CL 203.  
     
     
         40 . The method according to  claim 23 , wherein said ligand is a member selected from the group consisting of human rhinovirus, major group receptor virus, lymphocyte-associated antigen-1 (LFA-1) and  Plasmodium falciparum .  
     
     
         41 . A pharmaceutical composition comprising a pharmaceutically acceptable solvent, diluent, adjuvant or a carrier, and, as the active ingredient, an effective amount of a polypeptide according to  claim 1 .  
     
     
         42 . A method for inducing irreversible uncoating of human rhinovirus, said method comprising contacting said human rhinovirus with ICAM-1 or a tICAM fragment thereof.  
     
     
         43 . A method of irreversibly inhibiting infectivity of a mammalian cell by a human rhinovirus, said method comprising contacting said human rhinovirus with ICAM-1 or a tICAM fragment thereof under conditions which allow the ICAM-1 or tICAM to bind to said rhinovirus; thereby stimulating irreversible uncoating of said rhinovirus.