Short-form human md-2 as a negative regulator of toll-like receptor 4 signaling

Abstract

The present invention is based on a novel, alternatively spliced human isoform of MD-2 (MD-2s). In addition, the present invention relates to modified MD-2 proteins, wherein one or more tyrosine residues have been mutated to phenylalanine. In various embodiments, the invention relates to methods and kits for preventing, reducing the likelihood of developing and/or treating various conditions using MD-2s. The invention also describes methods of determining the risk of a subject to various conditions.

1 - 6 . (canceled) 
     
     
         7 . A method of inhibiting toll-like receptor 4 signaling (“TLR4”), inhibiting lipopolysaccharide (“LPS”) signaling, treating a condition mediated by TLR4 signaling, or reducing a likelihood of developing a condition mediated by TLR4 signaling in a subject in need thereof, comprising:
 providing a purified polypeptide comprising an amino acid sequence that is at least 80% identical to SEQ ID NO:1 or SEQ ID NO:2; and 
 administering the polypeptide to the subject to inhibit TLR4 signaling, inhibit LPS signaling, treat the condition mediated by TLR4 signaling, or reduce the likelihood of developing the condition mediated by TLR4 signaling. 
 
     
     
         8 . The method of  claim 7 , wherein the condition mediated by TLR signaling is selected from the group consisting of sepsis, septic shock, inflammation, gram negative bacterial infection, gram negative bacterial lung infection, immune response, atherosclerosis and combinations thereof. 
     
     
         9 - 40 . (canceled) 
     
     
         41 . The method of  claim 7 , wherein the polypeptide comprises the amino acid sequence as disclosed by SEQ ID NO:1 or SEQ ID NO:2. 
     
     
         42 . The method of  claim 7 , wherein the polypeptide is glycosylated. 
     
     
         43 . The method of  claim 7 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:2 and 1-20 conservative amino acid substitutions. 
     
     
         44 . The method of  claim 7 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:2 and 1-20 amino acid insertions, deletions and/or substitutions. 
     
     
         45 . A method of inhibiting Lipopolysaccharide (“LPS”) induced lung inflammation in a subject in need thereof, comprising:
 providing a purified polypeptide comprising an amino acid sequence that is at least 85% identical to SEQ ID NO:1 or SEQ ID NO:2; and 
 administering the polypeptide to the subject to inhibit LPS induced lung inflammation. 
 
     
     
         46 . The method of  claim 45 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 1. 
     
     
         47 . The method of  claim 46 , wherein the polypeptide is glycosylated. 
     
     
         48 . The method of  claim 45 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO:2. 
     
     
         49 . The method of  claim 48 , wherein the polypeptide is glycosylated. 
     
     
         50 . The method of  claim 45 , wherein the purified polypeptide comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:1 or SEQ ID NO:2. 
     
     
         51 . The method of  claim 45 , wherein the purified polypeptide comprising an amino acid sequence that is at least 95% identical to SEQ ID NO:1 or SEQ ID NO:2. 
     
     
         52 . The method of  claim 45 , wherein the purified polypeptide comprising an amino acid sequence that is at least 96% identical to SEQ ID NO:1 or SEQ ID NO:2. 
     
     
         53 . The method of  claim 45 , wherein the purified polypeptide comprising an amino acid sequence that is at least 97% identical to SEQ ID NO:1 or SEQ ID NO:2. 
     
     
         54 . The method of  claim 45 , wherein the purified polypeptide comprising an amino acid sequence that is at least 98% identical to SEQ ID NO:1 or SEQ ID NO:2. 
     
     
         55 . The method of  claim 45 , wherein the purified polypeptide comprising an amino acid sequence that is at least 99% identical to SEQ ID NO:1 or SEQ ID NO:2.