DNA encoding tumor necrosis factor inhibitory protein and its use

Abstract

Recombinant DNA encoding Tumor Necrosis Factor (TNF) Inhibitory Protein, or an active fragment thereof, is obtained. The TNF Inhibitory Protein has the ability to inhibit: (a) the binding of TNF to its receptors, and (b) the cytotoxic effect of TNF. TNF Inhibitory Protein and salts, functional derivatives and active fractions thereof can be used to antagonize the deleterious effects of TNF by eliminating TNF from the body, and to reduce the cytotoxic activity of TNF by binding to TNF and thereby to inhibit the binding of TNF to its receptors. The DNA may be in an expression vector. Host cells transfected with such an expression vector may be used to produce the TNF Inhibitory Protein.

1 . An isolated DNA molecule comprising a nucleotide sequence encoding a fraction of a naturally occurring human polypeptide comprising the amino acid sequence: Asp-Ser-Val-Cys-Pro-Gln-Gly-Lys-Tyr-Ile-His-Pro-Gln-X-Asn-Ser (SEQ ID NO:1) wherein X is an unidentified amino acid residue, and said polypeptide has the ability to interact with TNF in such a manner as to: 
 (a) inhibit the binding of TNF to a TNF receptor; and    (b) inhibit the cytotoxic effect of TNF;    said sequence having a sufficient length to serve as an oligonucleotide probe.    
     
     
         3 . An isolated DNA molecule comprising a nucleotide sequence encoding a fraction of a naturally occurring human polypeptide which comprises the amino acid sequence: 
 Asp-Ser-Val-Cys-Pro-Gln-Gly-Lys-Tyr-Ile-His-Pro-Gln-X-Asn-Ser (SEQ ID NO:1)    wherein X is an unidentified amino acid residue and said polypeptide has the ability to interact with TNF in such a manner as to inhibit the binding of TNF to a TNF receptor and to inhibit the cytotoxic effect of TNF; 
 said sequence being of sufficient length to encode a fragment of said polypeptide of sufficient length to serve as an immunogen for raising antibodies against said polypeptide when fused to protein A.  
   
     
     
         3 . An isolated DNA molecule comprising the DNA sequence of SEQ ID NO:2 or a fragment of the sequence SEQ ID NO:2 having a sufficient length to serve as a DNA probe.  
     
     
         4 . An isolated DNA molecule comprising the DNA sequence of SEQ ID NO:2 or a fragment of the sequence SEQ ID NO:2 having a sufficient length to encode a fragment of a polypeptide having the ability to interact with TNF in such a manner as to inhibit the binding of TNF to a TNF receptor and to inhibit the cytotoxic effect of TNF, said polypeptide fragment being of sufficient length to serve as an immunogen for raising antibodies against said polypeptide when fused to protein A.