Chemokine beta-7 variants

Abstract

The present invention relates to deletion and substitution mutant polypeptides of human chemokine β-7 (Ckβ-7), as well as nucleic acid molecules encoding such polypeptides and processes for producing such polypeptides using recombinant techniques. In one aspect, the invention also relates to uses of the full-length and mature forms of Ckβ-7, as well as deletion and substitution mutants, in medical treatment regimens. In particular, the Ckβ-7 polypeptides described herein may be employed to treat a variety of conditions, including rheumatoid arthritis, inflammation, respiratory diseases, allergy, and IgE-mediated allergic reactions.

1 . An isolated chemokine β7 C-terminal deletion mutant selected from the group consisting of: 
 (a) a polypeptide having an amino acid sequence selected from the group consisting of 1-70. 1-71, 1-72, 1-73, 1-74, 1-75, 1-76, 1-77, 1-78, 1-79, 1-80, 1-81, 1-82, 1-83, 1-84, 1-85, 1-86, 1-87 and 1-88 of SEQ ID NO:2;    (b) a polypeptide having an amino acid sequence selected from the group consisting of 21-70, 21-71, 21-72, 21-73, 21-74, 21-75, 21-76, 21-77, 21-78, 21-79, 21-80, 21-81, 21-82, 21-83, 21-84, 21-85, 21-86, 21-87, and 21-88 of SEQ ID NO:2;    (c) a polypeptide having an amino acid sequence of (a) plus a methionine residue at the N-terminus;    (d) a polypeptide having an amino acid sequence of (b) plus a methionine residue at the N-terminus;    (e) a substitution variant having an amino acid sequence of (a) except for one or more amino acid substitutions; wherein the amino acid sequence of said variant is at least 90% identical to said amino acid sequence of (a) and said variant inhibits chemokine induced calcium flux in eosinophils;    (f) a substitution variant having an amino acid sequence of (b) except for one or more amino acid substitutions; wherein the amino acid sequence of said variant is at least 90% identical to said amino acid sequence of (b) and said variant inhibits chemokine induced calcium flux in eosinophils;    (g) a substitution variant having an amino acid sequence of (c) except for one or more amino acid substitutions; wherein the amino acid sequence of said variant is at least 90% identical to said amino acid sequence of (c) and said variant inhibits chemokine induced calcium flux in eosinophils; and    (h) a substitution variant having an amino acid sequence of (d) except for one or more amino acid substitutions; wherein the amino acid sequence of said variant is at least 90% identical to said amino acid sequence of (d) and said variant inhibits chemokine induced calcium flux in eosinophils.    
     
     
         2 . An isolated chemokine β-7 N-terminal and C-terminal deletion mutant selected from the group consisting of: 
 (a) a polypeptide having an amino acid sequence selected from the group consisting of 21-88, 22-88, 23-88, 24-88, 25-88, 26-88, 27-88, 28-88, 29-88, 30-88, 21-87, 22-87, 23-87, 24-87, 25-87, 26-87, 27-87, 28-87, 29-87, 30-87, 21-86, 22-86, 23-86, 24-86, 25-86, 26-86, 27-86, 28-86, 29-86, 30-86, 21-85, 22-85, 23-85, 24-85, 25-85, 26-85, 27-85, 28-85, 29-85, 30-85, 21-84, 22-84, 23-84, 24-84, 25-84, 26-84, 27-84, 28-84, 29-84, 30-84, 21-83, 22-83, 23-83, 24-83, 25-83, 26-83, 27-83, 28-83, 29-83, 30-83, 21-82, 22-82, 23-82, 24-82, 25-82, 26-82, 27-82, 28-82, 29-82, 30-82, 21-81, 22-81, 23-81, 24-81, 25-81, 26-81, 27-81, 28-81, 29-81, 30-81, 21-80, 22-80, 23-80, 24-80, 25-80, 26-80, 27-80, 28-80, 29-80, 30-80, 21-79, 22-79, 23-79, 24-79, 25-79, 26-79, 27-79, 28-79, 29-79, 30-79, 21-78, 22-78, 23-78, 24-78, 25-78, 26-78, 27-78, 28-78, 29-78, 30-78, 21-77, 22-77, 23-77, 24-77, 25-77, 26-77, 27-77, 28-77, 29-77, 30-77, 21-76, 22-76, 23-76, 24-76, 25-76, 26-76, 27-76, 28-76, 29-76, 30-76, 21-75, 22-75, 23-75, 24-75, 25-75, 26-75, 27-75, 28-75, 29-75, 30-75, 21-74, 22-74, 23-74, 24-74, 25-74, 26-74, 27-74, 28-74, 29-74, 30-74, 21-73, 22-73, 23-73, 24-73, 25-73, 26-73, 27-73, 28-73, 29-73, 30-73, 21-72, 22-72, 23-72, 24-72, 25-72, 26-72, 27-72, 28-72, 29-72, 30-72, 21-71, 22-71, 23-71, 24-71, 25-71, 26-71, 27-71, 28-71, 29-71, 30-71, 21-70, 22-70, 23-70, 24-70, 25-70, 26-70, 27-70, 28-70, 29-70 and 30-70 of SEQ ID NO:2;    (b) a polypeptide having an amino acid sequence of (a) plus a methionine residue at the N-terminus;    (c) a substitution variant having an amino acid sequence of (a) except for one or more amino acid substitutions; wherein the amino acid sequence of said variant is at least 90% identical to said amino acid sequence of (a) and said variant inhibits chemokine induced calcium flux in eosinophils; and    (d) a substitution variant having an amino acid sequence of (b) except for one or more amino acid substitutions; wherein the amino acid sequence of said variant is at least 90% identical to said amino acid sequence of (b) and said variant inhibits chemokine induced calcium flux in eosinophils.    
     
     
         3 . An isolated nucleic acid molecule which encodes a polypeptide of  claim 1 .  
     
     
         4 . A method for making a recombinant vector comprising inserting the nucleic acid molecule of  claim 3  into a vector.  
     
     
         5 . A recombinant vector produced by the method of  claim 4 .  
     
     
         6 . A method of making a recombinant host cell comprising introducing the recombinant vector of  claim 5  into a host cell.  
     
     
         7 . A recombinant host cell produced by the method of  claim 6 .  
     
     
         8 . A method for producing a polypeptide comprising culturing the host cell of  claim 7  under conditions such that said polypeptide is expressed and recovering said polypeptide.  
     
     
         9 . A method for inhibiting chemokine induced calcium flux in eosinophils in an individual comprising administering to the individual an effective amount of a chemokine β-7 polypeptide selected from the group consisting of: 
 (a) a polypeptide comprising amino acids 30 to 70 in SEQ ID NO:2;    (b) a polypeptide comprising the amino acid sequence of (a) plus a methionine residue at the N-terminus;    (c) a substitution variant comprising the amino acid sequence of (a) except for one or more amino acid substitutions; wherein the amino acid sequence of said variant is at least 90% identical to said amino acid sequence of (a); and    (d) a substitution variant comprising the amino acid sequence of (b) except for one or more amino acid substitutions; wherein the amino acid sequence of said variant is at least 90% identical to said amino acid sequence of (b);    wherein said chemokine β-7 polypeptide is administered in admixture with a pharmaceutically acceptable carrier.    
     
     
         10 . The method of  claim 9 , wherein said chemokine β-7 polypeptide inhibits eosinophil chemotaxis.  
     
     
         11 . The method of  claim 9 , wherein said chemokine β-7 polypeptide is administered for treating diseases and disorders selected from the group consisting of: inflammation, rheumatoid arthritis, allergic reactions, dermatitis, chronic urticaria, adult respiratory distress syndrome, asthma, idiopathic hyper-eosinophilic syndrome, eosinophilic myositis, eosinophilic fascitis, rhinitis and infectious diseases.