Method and reagent for diagnosis and/or evaluation of progression of graft-versus-host disease

Abstract

Disclosed is a method of diagnosing graft-versus-host disease, comprising measuring the level of CCL8 protein in a sample obtained from a subject as an indicator for the diagnosis or course of graft-versus-host disease. Also a diagnostic reagent for graft-versus-host disease comprising an anti-CCL8 antibody is disclosed. The method of the present invention enables diagnosis of the onset of graft-versus-host disease and monitoring of the progress, in particular, differential diagnosis between graft-versus-host disease and an infectious disease. The present invention also provides a method for treating graft-versus-host disease utilizing the anti-CCL8 antibody.

1 . A method for diagnosing graft-versus-host disease, comprising:
 measuring the level of CCL8 protein in a first sample obtained from a human subject or animal subject at a first time point;   measuring the level of CCL8 protein in a second sample obtained from said human subject or animal subject at a second, later time point;   comparing the levels of CCL8 protein to thereby diagnose the development of graft-versus-host disease;   wherein increased level of CCL8 protein in the second sample indicates the development of graft-versus-host disease.   
     
     
         2 . The method according to  claim 1 , wherein the diagnosis of graft-versus-host disease is made before clinical manifestations of graft-versus-host disease. 
     
     
         3 . The method according to  claim 1 , wherein the level of CCL8 protein is measured using an anti-CCL8 antibody. 
     
     
         4 . The method according to  claim 1 , wherein the level of CCL8 protein is measured using a method selected from mass spectrometry, high-performance liquid chromatography, and two-dimensional electrophoresis. 
     
     
         5 - 8 . (canceled) 
     
     
         9 . A method for evaluating therapeutic efficacy of graft-versus-host disease, comprising
 treating a human subject or animal subject in need of therapy for graft-versus-host disease;   measuring the level of CCL8 protein in a first sample obtained from said human subject or animal subject at a first time point before or after the therapy has begun;   measuring the level of CCL8 protein in a second sample obtained from said human subject or animal subject at a second, later time point after the therapy has begun;   comparing the levels of CCL8 protein to thereby evaluate the therapeutic efficacy of the treatment for graft-versus-host disease;   wherein increased level of CCL8 protein in the second sample indicates the therapy is not effective for treatment of graft-versus-host disease, and wherein decreased level of CCL8 protein in the second sample indicates the therapy is effective for treatment of graft-versus-host disease.   
     
     
         10 . The method according to  claim 9 , wherein the level of CCL8 protein is measured using an anti-CCL8 antibody. 
     
     
         11 . The method according to  claim 9 , wherein the level of CCL8 protein is measured using a method selected from mass spectrometry, high-performance liquid chromatography, and two-dimensional electrophoresis. 
     
     
         12 . The method according to  claim 2 , wherein the level of CCL8 protein is measured using an anti-CCL8 antibody. 
     
     
         13 . The method according to  claim 2 , wherein the level of CCL8 protein is measured using a method selected from mass spectrometry, high-performance liquid chromatography, and two-dimensional electrophoresis. 
     
     
         14 . A method of diagnosing graft-versus-host disease, comprising:
 identifying a human subject who is treated with hematopoietic stem cell transplantation;   measuring the level of CCL8 protein in a first sample obtained from the human subject at a first time point and in a second sample obtained from the human subject at a second, later time point, by:
 obtaining an anti-CCL8 antibody reagent and using the anti-CCL8 antibody reagent to measure the level of CCL8 protein, or 
 using a method selected from mass spectrometry, high-performance liquid chromatography, and two-dimensional electrophoresis to measure the level of CCL8 protein; 
   and   comparing the levels of CCL8 protein so obtained wherein increased level of CCL8 protein in the second sample indicates the development of graft-versus-host disease.   
     
     
         15 . A method of evaluating the development of clinical manifestations of graft-versus-host disease, comprising:
 identifying a human subject who is treated with hematopoietic stem cell transplantation;   measuring the level of CCL8 protein in a first sample obtained from the human subject at a first time point and in a second sample obtained from the human subject at a second, later time point, by:
 obtaining an anti-CCL8 antibody reagent and using the anti-CCL8 antibody reagent to measure the level of CCL8 protein, or 
 using a method selected from mass spectrometry, high-performance liquid chromatography, and two-dimensional electrophoresis to measure the level of CCL8 protein; 
   and   comparing the levels of CCL8 protein so obtained wherein increased level of CCL8 protein in the second sample indicates the development of clinical manifestations of graft-versus-host disease.   
     
     
         16 . A method of evaluating the progression of clinical manifestations of graft-versus-host disease, comprising:
 identifying a human subject who has graft-versus-host disease;   measuring the level of CCL8 protein in a first sample obtained from the human subject at a first time point and in a second sample obtained from the human subject at a second, later time point, by:
 obtaining an anti-CCL8 antibody reagent and using the anti-CCL8 antibody reagent to measure the level of CCL8 protein, or 
 using a method selected from mass spectrometry, high-performance liquid chromatography, and two-dimensional electrophoresis to measure the level of CCL8 protein; 
   and   comparing the levels of CCL8 protein so obtained wherein increased level of CCL8 protein in the second sample indicates the progression of clinical manifestations of graft-versus-host disease.   
     
     
         17 . A method of diagnosing graft-versus-host disease made before clinical manifestations of graft-versus-host disease, comprising:
 identifying a human subject who is treated with hematopoietic stem cell transplantation but is not yet showing any clinical manifestations of graft-versus-host disease;   measuring the level of CCL8 protein in a first sample obtained from the human subject at a first time point and in a second sample obtained from the human subject at a second, later time point, by:
 obtaining an anti-CCL8 antibody reagent and using the anti-CCL8 antibody reagent to measure the level of CCL8 protein, or 
 using a method selected from mass spectrometry, high-performance liquid chromatography, and two-dimensional electrophoresis to measure the level of CCL8 protein; 
   and   comparing the levels of CCL8 protein so obtained wherein increased level of CCL8 protein in the second sample indicates the development of graft-versus-host disease irrespective of the presence of clinical manifestations of graft-versus-host disease.   
     
     
         18 . A method of initiating treatment of graft-versus-host disease conducted before clinical manifestations of graft-versus-host disease, comprising:
 identifying a human subject who is treated with hematopoietic stem cell transplantation but is not yet showing any clinical manifestations of graft-versus-host disease;   measuring the level of CCL8 protein in a first sample obtained from the human subject at a first time point and in a second sample obtained from the human subject at a second, later time point, by:
 obtaining an anti-CCL8 antibody reagent and using the anti-CCL8 antibody reagent to measure the level of CCL8 protein, or 
 using a method selected from mass spectrometry, high-performance liquid chromatography, and two-dimensional electrophoresis to measure the level of CCL8 protein; 
   and   comparing the levels of CCL8 protein so obtained wherein increased level of CCL8 protein in the second sample indicates the development of graft-versus-host disease irrespective of the presence of clinical manifestations of graft-versus-host disease; and   beginning treatment for graft-versus-host disease based on the increased levels in the second sample.   
     
     
         19 . A method of diagnosing graft-versus-host disease, comprising:
 identifying a human subject who is treated with hematopoietic stem cell transplantation; and   measuring the level of CCL8 protein in a sample obtained from the human subject, by:
 obtaining an anti-CCL8 antibody reagent and using the anti-CCL8 antibody reagent to measure the level of CCL8 protein, or 
 using a method selected from mass spectrometry, high-performance liquid chromatography, and two-dimensional electrophoresis to measure the level of CCL8 protein; 
   wherein the level of CCL8 protein of at least 52.0 pg/ml indicates the development of graft-versus-host disease.   
     
     
         20 . A kit for diagnosis of graft-versus-host disease, comprising
 an antibody reagent for measuring the level of a CCL8 protein,   reagents for use with the antibody reagent, and   means for detecting the level of the CCL8 protein.