Method for Monitoring the Immune Response and Predicting Clinical Outcomes in transplant recipients

Abstract

Methods for monitoring the immune response and predicting clinical outcomes for patients on immunosuppressive drugs (such as transplant patients) are provided. The methods are based on the measurement of an intracellular metabolic marker in lymphocytes (such as ATP) as an indicator of a patient's immune response.

1 - 49 . (canceled) 
   
   
       50 . A method of assessing an immune response to an antigen stimulant in a patient, comprising the steps of:
 incubating a blood sample from said patient for 0.5-48 hrs, at least a first portion of said blood sample including an antigen stimulant, and at least a second portion of said blood sample being non-stimulated;   selectively separating a subset of lymphocytes from said first portion and from said second portion of said blood sample, said subset of lymphocytes being selected from CD3, CD4, CD19, and CD56 cells, said selectively separating step being performed with a substrate having an antibody specific for said subset of lymphocytes;   lysing said subset of lymphocytes from each of said first portion and said second portion of said blood sample to produce a lysate containing adenosine triphosphate (ATP);   quantifying ATP in said lysate for said subset of lymphocytes from said first portion and said second portion of said blood sample; and   determining if said patient has been previously exposed to said antigen stimulant based on a difference between said ATP in said lysate for said subset of lymphocytes from said first portion and said second portion of said blood sample.   
   
   
       51 . The method of  claim 50  wherein said quantifying step is performed with a luciferin:luciferase assay. 
   
   
       52 . The method of  claim 50  wherein said substrate used in said selectively separating step is paramagnetic beads. 
   
   
       53 . The method of  claim 50  wherein said lymphocytes are CD3 lymphocytes. 
   
   
       54 . A method of monitoring an immune response in a patient receiving at least one immunosuppresive drug, comprising the steps of:
 at first and second time periods, determining a value of an immune response for said patient which reflects activation of one of CD3, CD4, CD 19, or CD56 lymphocytes in a whole blood sample stimulated by exposure to a mitogen or antigen stimulant wherein said value is measured in terms of adenosine triphosphate (ATP) present in cell lysate of said one or CD3, CD4, CD19 or CD 56 lymphocytes isolated from said blood sample;   comparing said value determined at said first time period with said value determined at said second time period, and if there is a decrease of at least about 50 ng/ml ATP, determining that said patient is at risk of being overmedicated with said one or more immunosuppressive drugs.   
   
   
       55 . The method of  claim 54  wherein said patient has at least one of Crohn's disease, cancer, human immunodeficiency virus, inflammation, rheumatoid arthritis, or lupus erythmatosus. 
   
   
       56 . The method of  claim 54  wherein said patient is an organ or non-organ transplant patient.