I-CAM related protein

Abstract

DNA sequences encoding a novel human intercellular adhesion molecule polypeptide (designated “ICAM-R”) and variants thereof are disclosed along with methods and materials for production of the same by recombinant procedures. Antibodies substances specific for ICAM-R and variants thereof are also disclosed as useful in both the isolation of ICAM-R from natural cellular sources and the modulation of ligand/receptor binding reactions involving ICAM-R.

1 . A purified and isolated polynucleotide encoding ICAM-R polypeptide or a variant thereof possessing a ligand/receptor binding biological activity or immunological property specific to ICAM-R.  
     
     
         2 . The polynucleotide of  claim 1  which is a DNA sequence.  
     
     
         3 . The DNA sequence according to  claim 2  which is a cDNA sequence or a biological replica thereof.  
     
     
         4 . The DNA sequence according to  claim 2  which is a genomic DNA sequence or a biological replica thereof.  
     
     
         5 . The DNA sequence of  claim 3  further including an endogenous expression control DNA sequence.  
     
     
         6 . The DNA sequence according to claim  0 . 2  which is a wholly or partially chemically synthesized DNA sequence or a biological replica thereof.  
     
     
         7 . A DNA vector comprising a DNA sequence according to  claim 2 .  
     
     
         8 . The vector of  claim 7  which is plasmid pCDNA-1-neo-ICAM-R.  
     
     
         9 . The vector of  claim 7  wherein said DNA sequence is operatively linked to an expression control DNA sequence.  
     
     
         10 . A host cell stably transformed or transfected with a DNA sequence according to  claim 2  in a manner allowing the expression in said host cell of ICAM-R polypeptide or a variant thereof possessing a ligand/receptor binding biological activity or immunological property specific to ICAM-R.  
     
     
         11 . A method for producing ICAM-R polypeptide or a variant thereof possessing a ligand/receptor binding biological activity or immunological property specific to ICAM-R, said method comprising growing a host cell according to  claim 10  in a suitable nutrient medium and isolating ICAM-R polypeptide or variant thereof from said cell or the medium of its growth.  
     
     
         12 . Purified and isolated ICAM-R polypeptide or a variant thereof possessing ligand/receptor binding a biological activity or immunological property specific to ICAM-R.  
     
     
         13 . The polypeptide of  claim 12  comprising residues −29 through 518 of SEQ ID NO: 1.  
     
     
         14 . The polypeptide of  claim 12  comprising residues 1 through 518 of SEQ ID NO: 1.  
     
     
         15 . The polypeptide of  claim 12  comprising residues 1 to 482 of SEQ ID NO: 1.  
     
     
         16 . The polypeptide of  claim 12  comprising residues 1 to 456 of SEQ ID NO: 1.  
     
     
         17 . The polypeptide of  claim 12  comprising residues 456 to 518 of SEQ ID NO: 1.  
     
     
         18 . The polypeptide of  claim 12  comprising residues 482 to 518 of SEQ ID NO: 1.  
     
     
         19 . The polypeptide of  claim 12  comprising a polypeptide selected from the group consisting of the following residues in SEQ ID NO: 1; 1 through 90; 91 through 187; 188 through 285; 286 through 387; and 388 through 481.  
     
     
         20 . The polypeptide of  claim 12  comprising a polypeptide selected from the group consisting of the following residues within. SEQ ID NO: 1; 24 through 71; 28 through 67; 110 through 161; 212 through 265; 307 through 346; and 394 through 433.  
     
     
         21 . An ICAM-R variant polypeptide according to  claim 12  in multimeric form.  
     
     
         22 . A water soluble polypeptide according to  claim 12 .  
     
     
         23 . An antibody substance specific for ICAM-R.  
     
     
         24 . A monoclonal antibody according to  claim 23 .  
     
     
         25 . A monoclonal antibody according to  claim 24  produced by hybridoma cell line 26E3D-1.  
     
     
         26 . A monoclonal antibody according to  claim 24  produced by hybridoma cell line 26I8F-2.  
     
     
         27 . A monoclonal antibody according to  claim 24  produced by hybridoma cell line 26I10E-2.  
     
     
         28 . A monoclonal antibody according to  claim 24  produced by hybridoma cell line 26H11C-2.  
     
     
         29 . A hybridoma cell line producing a monoclonal antibody according to  claim 24  selected from the group consisting of hybridoma cell lines designated 26E3D-1, 26I8F-2, 26I10E-2, and 26H11C-2.  
     
     
         30 . An anti-idiotypic antibody substance specific for the antibody substance according to  claim 23 .  
     
     
         31 . A humanized antibody substance according to claims  23  or  30 .  
     
     
         32 . A method for modulating ligand/receptor binding of ICAM-R comprising contacting ICAM-R with an antibody according to  claim 23  or  30 .  
     
     
         33 . The method of  claim 32  wherein ICAM-R ligand/receptor binding is inhibited.  
     
     
         34 . The method of  claim 32  wherein ICAM-R ligand/receptor binding is stimulated.  
     
     
         35 . The method of  claim 34  wherein the antibody is an antibody produced by a hybridoma selected from the group consisting of 26E3D-1, 26I8F-2, and 26H11C-2.  
     
     
         36 . A method for treating inflammation resulting from a response of the specific immune system in a mammalian subject comprising providing to a mammal in need of such treatment an amount of an antibody substance according to claims  23  or  30  sufficient to suppress inflammation.  
     
     
         37 . A method for treating inflammation resulting from a response of the nonspecific immune system in a mammalian subject comprising providing to a mammal in need of such treatment an amount of an antibody substance according to claims  23  or  30  sufficient to suppress inflammation.  
     
     
         38 . A method for monitoring an inflammatory disease state comprising detecting and quantifying at least one soluble ICAM-R fragment in the serum of a patient and comparing the quantities so determined to the serum quantity of said ICAM-R fragment in the absence of an inflammatory state.  
     
     
         39 . The method of  claim 38  wherein said detecting and quantifying steps comprise: contacting serum from a patient with a first monoclonal antibody specific for a first epitope of ICAM-R; reacting any ICAM-R fragment bound to said first monoclonal antibody with a second monoclonal antibody specific for a second epitope of ICAM-R; and quantifying the amount of second monoclonal antibody bound.  
     
     
         40 . The method of  claim 39  wherein said second monoclonal antibody is detectably labelled.  
     
     
         41 . A method for detecting the capacity of a cell to synthesize ICAM-R comprising hybridizing a detectable polynucleotide encoding ICAM-R or a fragment thereof with RNA of said cell.  
     
     
         42 . A method for detecting the capacity of a cell to synthesize ICAM-R comprising reacting an antibody substance according to  claim 23  with polypeptides produced by said cell.  
     
     
         43 . An antisense polynucleotide for a polynucleotide encoding ICAM-R.  
     
     
         44 . An antisense polynucleotide for a DNA specifying an endogenous expression control DNA sequence of ICAM-R.  
     
     
         45 . A hybrid fusion polypeptide comprising, at its amino terminal, an ICAM-R polypeptide or a variant thereof possessing a ligand/receptor binding, biological activity or immunological property specific to ICAM-R and, at its carboxy terminal, at least one constant domain of an immunoglobulin heavy chain or allelic variant thereof.  
     
     
         46 . A polynucleotide encoding a hybrid fusion protein according to  claim 45 .  
     
     
         47 . An immunogen suitable for use in eliciting the formation of an antibody monospecific for ICAM-R and selected from the group consisting of cells naturally expressing ICAM-R; recombinant host cells expressing ICAM-R or a variant thereof having an immunological reactivity specific for ICAM-R; ICAM-R or a variant thereof having an immunological reactivity specific for ICAM-R; and ICAM-R which is bound to an antibody which stimulates ICAM-R ligand/receptor binding.  
     
     
         48 . A DNA sequence encoding a polypeptide having a ligand/receptor biological or an immunological property specific for ICAM-R and selected from the group consisting of: 
 (a) the DNA sequence set out in SEQ ID NO: 2;    (b) a DNA which hybridizes under stringent conditions to the DNA of (a); and    (c) a DNA sequence which, but for the redundancy of the genetic code, would hybridize under stringent conditions to a DNA sequence of (a) or (b).