Diagnosing and monitoring inflammatory diseases by measuring complement components on white blood cells

Abstract

The invention is related to methods of diagnosing inflammatory diseases or conditions by determining levels of components of the complement pathway on the surface of white blood cells.

1 . A method for diagnosing an inflammatory disease in an individual, the method comprising,
 (a) quantitating, in a blood sample containing white blood cells from the individual, a level of a C4d and/or C3d component of the complement pathway on a surface of a T lymphocyte, B lymphocyte or monocyte in the sample, and   (b) comparing the level in (a) with a level of C4d and/or C3d with the component of the complement pathway on the surface of a control T lymphocyte, B lymphocyte or monocyte from an individual not having the inflammatory disease, wherein an increased level of the C4d and/or C3d component of the complement pathway diagnoses the inflammatory disease in the individual.   
     
     
         2 . The method of  claim 1 , wherein the method comprises quantitating, in the blood sample, a level of C4d and/or C3d on the surface of a T lymphocyte, wherein the T lymphocyte is isolated using an anti-CD3 antibody. 
     
     
         3 . The method of  claim 1 , comprising quantitating, in the blood sample, a level of a C4d component of the complement pathway on a surface of a T lymphocyte, B lymphocyte or monocyte in the sample. 
     
     
         4 . The method of  claim 1 , comprising quantitating, in the blood sample, a level of a C3d component of the complement pathway on a surface of a T lymphocyte, B lymphocyte or monocyte in the sample. 
     
     
         5 . The method of  claim 1 , wherein the inflammatory disease or condition is selected from the group consisting of scleroderma, rheumatoid arthritis, vasculitis, myositis, multiple sclerosis, gout, pre-eclampsia, serum sickness, cardiovascular disease, systemic lupus erythematosus (SLE) and hepatitis C virus infection. 
     
     
         6 . The method of  claim 3 , wherein the level of the C4d component of the complement pathway is quantitated using an antibody specific for the C4d component of the complement pathway. 
     
     
         7 . The method of  claim 6 , wherein the antibody specific for the C4d component of the complement pathway is labeled. 
     
     
         8 . The method of  claim 6 , wherein the antibody specific for the C4d component of the complement pathway is a monoclonal antibody. 
     
     
         9 . The method of  claim 3 , wherein the level of at least one other complement component is quantitated. 
     
     
         10 . The method of  claim 4 , wherein the level of the C3d component of the complement pathway is quantitated using an antibody specific for C3d component of the complement pathway. 
     
     
         11 . The method of  claim 10 , wherein the antibody specific for the C3d component of the complement pathway is labeled. 
     
     
         12 . The method of  claim 10 , wherein the antibody specific for the C3d component of the complement pathway is a monoclonal antibody. 
     
     
         13 . The method of  claim 9 , wherein the complement component is C1, C4, C3, C3, C1q, C1r, C1s, C4a, C4b, C2a, C2b, C4b2a, C3a, C3b, C4c, iC3b, C3i, C3dg, C5, C5b, C6, C7, C8, C9, C1inh, MASP2, CR1, DAF, MCP, CD59, C3aR, C1qR, CR2, CR3 or CR4. 
     
     
         14 . The method of  claim 9 , wherein the blood sample is treated with ethylenediaminetetraacetate (EDTA) to inhibit complement activation. 
     
     
         15 . A method for diagnosing an inflammatory disease in an individual of interest, the method comprising,
 (a) isolating lymphocytes from the individual of interest using fluorescence activated cell sorting;   (b) determining in the lymphocytes, a level of a C4d and/or C3d component of the complement pathway on surface of the lymphocytes by determining the mean fluorescence channel, and   (c) comparing the mean fluorescence channel in (b) with a mean fluorescence channel of lymphocytes from a control individual not having the inflammatory disease,   wherein an increased mean fluorescence channel of the lymphocytes from the individual of interest as compared to the mean fluorescence channel of the control individual diagnoses the inflammatory disease in the individual of interest.   
     
     
         16 . A computer readable medium, storing computer-executable instructions for implementing an evaluation tool using an automated system, wherein the automated system comprises memory and a processor, and wherein the evaluation tool evaluates complement component C4d deposits and/or complement component C3d deposits on surfaces of T lymphocytes, B lymphocytes or monocytes, the computer-executable instructions causing the processor to:
 receive data corresponding to complement component C4d and/or complement component C3d deposited on surfaces of T lymphocytes, B lymphocytes or monocytes,   store the data corresponding to the complement component C4d and/or complement component C3d in the memory of the automated system;   store the retrieved reference value in the memory of the automated system;   compare the received data with the reference value; and   store results of the comparing in the memory of the automated system.   
     
     
         17 . The computer readable medium of  claim 16 , wherein the evaluation tool evaluates complement component C3d deposits and/or complement component C4d deposits, and wherein data corresponding to complement component C3d and complement component C4d are received. 
     
     
         18 . The computer readable medium of  claim 16 , wherein the evaluation tool evaluates C4d deposits and/or C3d deposits on the surface of T lymphocytes. 
     
     
         19 . The computer readable medium of  claim 16 , wherein the computer readable medium is read by a digital computer that displays a determination if the complement component C3d deposits and the complement component C4d deposits are associated with an inflammatory condition 
     
     
         20 . The computer readable medium of  claim 17 , wherein the inflammatory condition is systemic lupus erythematosus.