Hematopoietic cell phenotyping using free circulating cellular markers

Abstract

The present invention provides methods of identifying cluster of differentiation (CD) marker phenotype for hematopoietic cells using multiple soluble CD markers circulating in bodily fluid. In particular aspects, the CD marker phenotype can be used to classify the tumor type of a patient having a proliferative disorder. In other aspects, treatment and disease progression can be monitored by measuring the levels of CD markers in bodily fluids of a patient over time.

1 . A method for identifying a cluster of differentiation (CD) phenotype of hematopoietic cells underlying a disease or disorder in an individual, said disease or disorder selected from the group consisting of a hematopoietic proliferative disorder, autoimmune disease, and infection, said method comprising, 
 determining a level of two or more CD markers circulating in a bodily fluid sample from said individual, wherein if only two CD markers are used, said two CD markers are not CD20 and CD52;    comparing said level of each of said CD markers to the level of the corresponding CD marker in a bodily fluid of a normal individual;    identifying the CD phenotype by assigning any individual CD marker having a level different from the level of said corresponding CD marker in the bodily fluid of a normal individual to the phenotype.    
   
   
       2 . A method according to  claim 1 , wherein said CD markers are not actively secreted or actively shed by the cells.  
   
   
       3 . A method according to  claim 1 , wherein said hematopoietic proliferative disorder is selected from the group consisting of lymphocytic leukemia, acute myelogenous leukemia, acute lymphoblastic leukemia, myelodysplastic syndrome, chronic myelomonocytic leukemia, juvenile myelomonocyte leukemia, multiple myeloma, lymphoma, T-cell chronic lymphocytic leukemia, prolymphocytic leukemia, hairy cell leukemia, follicular lymphoma, lymphoplasmocytic immunocytoma, plasma cell leukemia, T prolymphocytic leukemia, mycosis fungoides, large granular lymphocyte leukemia, and adult T cell leukemia.  
   
   
       4 . A method according to  claim 1 , wherein said autoimmune disease is selected from the group consisting of ulcerative colitis, polyarteritis, pemphigus complex and systemic lupus erythematosus.  
   
   
       5 . A method according to  claim 1 , wherein said infection is HIV and wherein said hematopoietic cells underlying the infection are T4 and T8 cells.  
   
   
       6 . A method according to  claim 1 , wherein said bodily fluid is selected from the group consisting of blood, plasma, serum, lymphatic fluid, cerebrospinal fluid, synovial fluid, urine, and saliva.  
   
   
       7 . A method according to  claim 1 , wherein said determining is accomplished using antibodies specific for the CD markers.  
   
   
       8 . A method according to  claim 1 , wherein said measuring is accomplished using flow cytometry.  
   
   
       9 . A method according to  claim 1 , wherein said determining is accomplished with an enzyme-linked immunosorbent assay.  
   
   
       10 . A method according to  claim 1 , wherein said CD markers are selected from the group consisting of CD3, CD4, CD5, CD7, CD8, CD10, CD11c, CD19, CD20, CD22, CD23, CD25, CD30, CD31, CD38, CD44, CD52, CD56, CD57, CD86, CD103, CD146, and CD163.  
   
   
       11 . A method according to  claim 1 , wherein said CD markers are selected from the group consisting CD20, CD23, CD30, CD31, CD44, CD52, CD86, CD146, and CD163.  
   
   
       12 . A method according to  claim 1 , wherein said CD markers are selected from the group consisting CD5, CD10, CD11c, CD19, CD20, CD22, CD23, CD25, CD38, and CD103.  
   
   
       13 . A method according to  claim 1 , wherein said CD markers are selected from the group consisting CD3, CD4, CD5, CD7, CD8, CD56, and CD57.  
   
   
       14 . A method according to  claim 1 , wherein said CD markers are selected from the group consisting CD20, CD52, and CD86.  
   
   
       15 . The method of  claim 1  wherein said hematopoietic proliferative disorder is a hematopoietic tumor.  
   
   
       16 . A method of classifying a hematopoietic tumor type via its cluster of differentiation (CD) marker phenotype in an individual for the purposes of prognosis or treatment of said individual, said method comprising, 
 determining a level of two or more CD markers circulating in a bodily fluid sample from said individual with a hematopoietic proliferative disorder, wherein if only two CD markers are used, said two CD markers are not CD20 and CD52;    comparing said level of each of said CD markers to the level of the corresponding CD marker in a bodily fluid of a normal individual;    identifying the CD phenotype by assigning any individual CD marker having a level larger than the level of said corresponding CD marker in the bodily fluid of a normal individual to the phenotype.    comparing the phenotype to the cell-surface CD marker phenotype of known hematopoietic tumor types to classify the tumor type in said individual with a hematopoietic proliferative disorder for prognosis or treatment of that disorder.    
   
   
       17 . A method according to  claim 16 , wherein said hematopoietic tumor type is selected from the group consisting of lymphocytic leukemia, acute myelogenous leukemia, acute lymphoblastic leukemia, myelodysplastic syndrome, chronic myelomonocytic leukemia, juvenile myelomonocyte leukemia, multiple myeloma, lymphoma, T-cell chronic lymphocytic leukemia, prolymphocytic leukemia, hairy cell leukemia, follicular lymphoma, lymphoplasmocytic immunocytoma, plasma cell leukemia, T prolymphocytic leukemia, mycosis fungoides, large granular lymphocyte leukemia, and adult T cell leukemia.  
   
   
       18 . A method according to  claim 16 , wherein said bodily fluid is selected from the group consisting of blood, plasma, serum, lymphatic fluid, cerebrospinal fluid, synovial fluid, urine, and saliva.  
   
   
       19 . A method according to  claim 16 , wherein said determining is accomplished using antibodies specific for the CD markers.  
   
   
       20 . A method according to  claim 16 , wherein said CD markers are selected from the group consisting of CD3, CD4, CD5, CD7, CD8, CD10, CD11c, CD19, CD20, CD22, CD23, CD25, CD30, CD31, CD38, CD44, CD52, CD56, CD57, CD86, CD103, CD146, and CD163.  
   
   
       21 . A method according to  claim 16 , wherein said CD markers are selected from the group consisting CD20, CD23, CD30, CD31, CD44, CD52, CD86, CD146, and CD163.  
   
   
       22 . A method according to  claim 16 , wherein said CD markers are selected from the group consisting CD5, CD10, CD11c, CD19, CD20, CD22, CD23, CD25, CD38, and CD103.  
   
   
       23 . A method according to  claim 16 , wherein said CD markers are selected from the group consisting CD3, CD4, CD5, CD7, CD8, CD56, and CD57.  
   
   
       24 . A method according to  claim 16 , wherein said CD markers are selected from the group consisting CD20, CD52, and CD86.  
   
   
       25 . A method of monitoring treatment or progression of a hematopoietic proliferative disorder in a patient, said method comprising, 
 obtaining a first bodily fluid sample from a patient having a hematopoietic proliferative disorder;    determining a level of two or more CD markers circulating in said first bodily fluid sample, wherein if only two CD markers are used, said two CD markers are not CD20 and CD52;    obtaining a second bodily fluid sample, wherein said second bodily fluid sample is obtained after said first bodily fluid sample;    determining a level of said two or more CD markers circulating in said second bodily fluid bodily fluid sample;    comparing the levels of said two or more CD markers in said first bodily fluid sample to the level of the corresponding CD marker in said second bodily fluid sample, wherein an increase in the levels in said second bodily fluid sample relative to said first bodily fluid sample is indicative of progression of the disease, or wherein a decrease in the levels in said second bodily fluid sample relative to said first bodily fluid sample is indicative of a regression of the disease.    
   
   
       26 . A method according to  claim 25 , wherein said hematopoietic proliferative disorder is selected from the group consisting of lymphocytic leukemia, acute myelogenous leukemia, acute lymphoblastic leukemia, myelodysplastic syndrome, chronic myelomonocytic leukemia, juvenile myelomonocyte leukemia, multiple myeloma, lymphoma, T-cell chronic lymphocytic leukemia, prolymphocytic leukemia, hairy cell leukemia, follicular lymphoma, lymphoplasmocytic immunocytoma, plasma cell leukemia, T prolymphocytic leukemia, mycosis fungoides, large granular lymphocyte leukemia, and adult T cell leukemia.  
   
   
       27 . A method according to  claim 25 , wherein said bodily fluid is selected from the group consisting of blood, plasma, serum, lymphatic fluid, cerebrospinal fluid, synovial fluid, urine, and saliva.  
   
   
       28 . A method according to  claim 25 , wherein said determining is accomplished using antibodies specific for the CD markers.  
   
   
       29 . A method according to  claim 25 , wherein said CD markers are selected from the group consisting of CD3, CD4, CD5, CD7, CD8, CD10, CD11c, CD19, CD20, CD22, CD23, CD25, CD30, CD31, CD38, CD44, CD52, CD56, CD57, CD86, CD103, CD146, and CD163.  
   
   
       30 . A method according to  claim 25 , wherein said CD markers are selected from the group consisting CD20, CD23, CD30, CD31, CD44, CD52, CD86, CD146, and CD163.  
   
   
       31 . A method according to  claim 25 , wherein said CD markers are selected from the group consisting CD5, CD10, CD11c, CD19, CD20, CD22, CD23, CD25, CD38, and CD103.  
   
   
       32 . A method according to  claim 25 , wherein said CD markers are selected from the group consisting CD3, CD4, CD5, CD7, CD8, CD56, and CD57.  
   
   
       33 . A method according to  claim 25 , wherein said CD markers are selected from the group consisting CD20, CD52, and CD86.