Treating arthritis by cleaving TNF receptors

Abstract

The biological effects of the cytokine TNF are mediated by binding to receptors on the surface of cells. This disclosure describes new proteins and polynucleotides that promote enzymatic cleavage and release of TNF receptors. Also provided are methods for identifying additional compounds that influence TNF receptor shedding. As the active ingredient in a pharmaceutical composition, the products of this invention increase or decrease TNF signal transduction, thereby alleviating the pathology of disease.

1 - 32 . (canceled)  
     
     
         33 . A method for treating arthritis or reducing inflammation in a subject, comprising cleaving TNF receptors from the surface of cells in the subject.  
     
     
         34 . The method of  claim 33 , wherein the TNF receptors are caused to be cleaved from the cells by administering to the subject a protein containing an amino acid sequence encoded in any of SEQ. ID NOs: 1-10, or fragment of any of said amino acid sequences that causes release of TNF receptor from human cells in which TNF is expressed.  
     
     
         35 . The method of  claim 33 , wherein the TNF receptors are caused to be cleaved from the cells by administering to the subject a protein encoded in a DNA that hybridizes to a DNA having the sequence of any of SEQ. ID NOs:1-10 at 30° C. in 6×SSC; 
 wherein the protein causes increased release of TNF receptor from human cells in which TNF is expressed.    
     
     
         36 . The method of  claim 33 , wherein the TNF receptors are caused to be cleaved from the cells by administering to the subject a protein containing an amino acid sequence encoded in SEQ. ID NO:9, or fragment thereof that causes release of TNF receptor from human cells in which TNF is expressed.  
     
     
         37 . The method of  claim 33 , wherein the TNF receptors are caused to be cleaved from the cells by administering to the subject a protein encoded in a DNA that hybridizes to a DNA having the sequence of SEQ. ID NO:9 at 30° C. in 6×SSC; 
 wherein the protein causes increased release of TNF receptor from human cells in which TNF is expressed.    
     
     
         38 . The method of  claim 33 , wherein the TNF receptors are caused to be cleaved from the cells by administering to the subject a protein containing an amino acid sequence that is at least 80% identical to SEQ. ID NO:9, 
 wherein the protein causes increased release of TNF receptor from human cells in which TNF is expressed.    
     
     
         39 . The method of  claim 33 , wherein the TNF receptors are caused to be cleaved from the cells by administering to the subject a composition comprising an isolated naturally occurring protein that causes TNF receptor cleavage, obtainable by a process comprising: 
 a) stimulating THP-1 cells with phorbol myristate acetate;    b) harvesting culture medium from the stimulated cells; and    c) isolating from the medium a protein that causes TNF receptor cleavage.    
     
     
         40 . The method of  claim 33 , wherein the TNF receptors are caused to be cleaved from the cells by administering to the subject the supernatant of THP-1 cells stimulated with phorbol myristate acetate.  
     
     
         41 . The method of  claim 39 , wherein the protein has been isolated by a method comprising ion exchange chromatography and separation by molecular weight.  
     
     
         42 . The method of  claim 39 , wherein release of TNF receptor caused by the protein is enhanced in the presence of Ca ++  and diminished in the presence of phenanthroline.  
     
     
         43 . The method of  claim 39 , wherein the protein has been obtained from THP-1, U-937, HL-60, ME-180, MRC-5, Raji, or K-562 cells or normal human monocytes.  
     
     
         44 . The method of  claim 33 , wherein the receptors cleaved from the surface of cells in the subject are human p75 TNF receptors.  
     
     
         45 . The method of  claim 33 , wherein the receptors cleaved from the surface of cells in the subject are human p55 TNF receptors.  
     
     
         46 . The method of  claim 34 , wherein the protein is at least about 90% homogeneous when analyzed by SDS polyacrylamide gel electrophoresis.  
     
     
         47 . The method of  claim 39 , wherein the protein is at least about 90% homogeneous when analyzed by SDS polyacrylamide gel electrophoresis.  
     
     
         48 . The method of  claim 34 , wherein the protein is administered systemically.  
     
     
         49 . The method of  claim 34 , wherein the protein is administered intravenously.  
     
     
         50 . The method of  claim 34 , wherein the protein is administered intramuscularly.  
     
     
         51 . The method of  claim 33 , whereby the subject is treated for sepsis.  
     
     
         52 . The method of  claim 33 , whereby the subject is treated for arthritis.  
     
     
         53 . The method of  claim 33 , whereby the subject is treated for rheumatoid arthritis.  
     
     
         54 . The method of  claim 33 , whereby the subject is treated for multiple sclerosis.