Human antibody specific for interleukin-1alpha

Abstract

Fully human monoclonal Abs includes (i) an antigen-binding variable region that exhibits very high binding affinity for IL-1α and (ii) a constant region that is effective at both activating the complement system though C1q binding and binding to several different Fc receptors.

What is claimed is: 
     
         1 . A purified human IgG1 mAb that specifically binds to human IL-1α, the mAb comprising a heavy chain covalently joined to a light chain. 
     
     
         2 . The purified human IgG1 monoclonal Ab of  claim 1 , wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 9 and the light chain comprises the amino acid sequence of SEQ ID NO:11. 
     
     
         3 . A set of isolated nucleic acids comprising a first nucleic acid encoding the heavy chain of a human IgG1 mAb that specifically binds to IL-1α, and a second nucleic acid encoding the light chain of the human IgG1 mAb that specifically binds to human IL-1α. 
     
     
         4 . The set of isolated nucleic acids of  claim 3 , wherein the first nucleic acid encodes the amino acid sequence of SEQ ID NO: 9 and the second nucleic acid encodes the amino acid sequence of SEQ ID NO:11. 
     
     
         5 . The set of nucleic acids of  claim 4 , wherein the first nucleic acid comprises the nucleotide sequence of SEQ ID NO: 10 and the second nucleic acid comprises the nucleotide sequence of SEQ ID NO:12. 
     
     
         6 . The set of isolated nucleic acids of  claim 3 , wherein the set of isolated nucleic acids is comprised within at least one expression vector. 
     
     
         7 . The set of isolated nucleic acids of  claim 4 , wherein the set of isolated nucleic acids is comprised within at least one expression vector. 
     
     
         8 . The set of isolated nucleic acids of  claim 5 , wherein the set of isolated nucleic acids is comprised within at least one expression vector. 
     
     
         7 . The set of isolated nucleic acids of  claim 3 , wherein the set of isolated nucleic acids is comprised within an isolated host cell. 
     
     
         8 . The set of isolated nucleic acids of  claim 4 , wherein the set of isolated nucleic acids is comprised within an isolated host cell. 
     
     
         9 . The set of isolated nucleic acids of  claim 8 , wherein the host cell is a mammalian cell. 
     
     
         10 . A method of killing a cell expressing human IL-1α, the method comprising the step of contacting the cell with a purified human IgG1 mAb that specifically binds to human IL-1α. 
     
     
         11 . The method of  claim 10 , wherein the human IgG1 mAb comprises a heavy chain comprising the amino acid sequence of SEQ ID NO. 9 and a light chain comprising the amino acid sequence of SEQ ID NO:11. 
     
     
         12 . A method of inhibiting migration of a cell through a basement membrane matrix, the method comprising the step of adding a purified mAb that specifically binds to human IL-1α to a mixture comprising the basement membrane matrix and the cell. 
     
     
         13 . The method of  claim 12 , wherein the purified mAb is a human IgG1 mAb comprising a heavy chain covalently joined to a light chain. 
     
     
         14 . The method of  claim 13 , wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 9 and a light chain comprises the amino acid sequence of SEQ ID NO:11. 
     
     
         15 . A method of inhibiting an IL-1α-induced increase in expression of ICAM-1 or E-selectin on the surface of a human endothelial cell, the method comprising the step of adding a purified mAb that specifically binds to human IL-1α to a mixture comprising the endothelial cell and IL-1α. 
     
     
         16 . The method of  claim 15 , wherein the purified mAb is a human IgG1 mAb comprising a heavy chain covalently joined to a light chain. 
     
     
         17 . The method of  claim 16 , wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 9 and a light chain comprises the amino acid sequence of SEQ ID NO:11. 
     
     
         18 . A method of tracking inflammation in a human subject previously subjected to the steps of: obtaining from the subject a first sample of peripheral blood mononuclear cells at a first time; contacting the first sample with a purified mAb that specifically binds to human IL-1α; and determining the percent of cells in the first sample that bind the monoclonal Ab, the method comprising the steps of:
 (a) obtaining from the subject a second sample of peripheral blood mononuclear cells at a second time; 
 (b) contacting the second sample with the purified mAb that specifically binds to human IL-1α; 
 (c) determining the percent of cells in the second sample that bind the monoclonal Ab; and 
 (d) comparing the percent of cells in the first sample that bind the mAb to the percent of cells in the second sample that bind the monoclonal Ab. 
 
     
     
         19 . The method of  claim 18 , wherein the purified mAb is a human IgG1 mAb comprising a heavy chain covalently joined to a light chain. 
     
     
         20 . The method of  claim 19 , wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 9 and a light chain comprises the amino acid sequence of SEQ ID NO:11. 
     
     
         21 . A method comprising the steps of:
 (a) enriching a biological sample obtained from a human subject using a filter that separates molecules according to molecular weight into a first fraction comprising intact IgG complexed with IL-1α and second fraction comprising molecule less than 100 Kda; and   (b) quantifying the amount of IL-1α in the first fraction.   
     
     
         22 . The method of  claim 21 , comprising the steps of:
 (a) enriching a sample of plasma obtained from a human subject using a filter that separates molecules according to molecular weight into a first fraction comprising intact IgG complexed with IL-1α and second fraction comprising molecules less than 100 Kda;   (b) adding the first fraction to a substrate comprising immobilized anti-human IgG Abs under conditions that allow IgG in the first fraction to specifically bind the anti-human IgG Abs immobilized on the substrate;   (c) washing the substrate to remove material in the first fraction that does not specifically bind the immobilized anti-human IgG Abs;   (d) contacting the substrate washed in step (c) with an Ab that specifically binds human IL-1α under conditions that allows the Ab that specifically binds human IL-1α to specifically bind any human IL-1α bound to the substrate;   (e) washing the substrate to remove any of the Ab that specifically binds human IL-1α that is not bound to the substrate; and   (f) quantifying the amount of Ab that specifically binds human IL-1α remaining bound to the substrate after step (e).