Clinical Application Of Immune Function Assay Through CD4~+T-Lymphocyte Adenosine Triphosphate(atp) Level In Renal Transplantation

Background:Management of renal transplant recipients require immunosuppression to avoidgraft rejection. The short-term survival of the graft has been greatly improved,while it isregrettable that rejection is still an obstacle to long-term survival of allograft.If allograftsurvival is improved by strengthening immunosuppression, patients will then be at risk ofinfection and cancer.In order to balance immunosuppression insufficiency and overdose,,itbecome an important issue in transplant work to assess the immune status of the organtransplant recipients.Most of the immunosuppresants which are used in transplantationeffect by restrain the active and effective function of the T cells.CD4~+T-lymphocyte Thefunction of immune cell depends on the ATP supply,so detection of the concentration ofATP can reflect the function of CD4~+T-lymphocyte. Cylex-ImmuknowTMtechnology hasbeen approved by FDA,whice is widely used in clinic work.It was used to measure theintracellular ATP concertration of CD4~+T-lymphocyte stimulated by PHA to reflect theimmune function.The cell-mediated immunity status and its relative factors in renalrecipients and healthy adults were analyzed.Objective:1.To explore the use of Cylex-ImmuknowTMtechnology to monitor kidneytransplantation postoperative immune in clinical application.2.Useing Cylex-ImmuknowTMtechnology moitoring intracellular ATP concertrationof CD4~+T-lymphocyte whice stimulated by PHA, in order to know the renal recipientsimmune state before and after operation..3.Using the Cylex-ImmuknowTMtechnology moitoring the ATP of renal transplantrecipients in the occurrence of acute rejection,lung infections and effective treatment,and exploreing the role this approach in prediction of rejection and infection..4.Using the Cylex-ImmuknowTMtechnology moitoring the cell-mediated immunityfunction after basiliximab induction treatment,and exploreing the impact ofbasiliximab on cell-mediated immunity function.5.In analysis of the correlation between ATP value and the CSA/FK506plasmaconcentration,and exploreing immuknow technology is a comprehensive assessment of therole of the state of the cellular immune function in individual therapy.Method:By using the Cylex-immuknow assay,immune cell function was assessed bymeasuring ATP content of CD4~+T cell in healthy people,renal transplant recipientspre-and post-transplant respectively.Between October2010and February2012,80renaltransplant recipients and25healthy people were studied,the blood samples were collectedat Department of Urology,General Hosipital of Jinan Military Region. Routine tripleimmunosuppresant was applied,while20patients were treated with basiliximabinduction.Immuknow test was performed either on a protocol timing (pretransplant,7,15,21,30days post-transplant,acute rejection, infection, acute rejection and infection effectivetreatment), in addition determination of the intracellular ATP concertration of CD4~+T-lymphocyte in healthy people.Determination of the corresponding plasmaconcentration(FK506/CSA). Immuknow test were as follow:1.cell stimulation2.cellselection and ATP release.3.ATP measurement and data analysis.Then we carry outstatistic analysis on the base of those datas.Results:The average ATP concentration of healthy adults and patients before renaltransplantation were (441.12±88.76) ng/ml and (385.54±121.76)ng/mlrespectively(P<0.05).The ATP concentration of patients after renal transplantation(7,15,21,30days)were(180.68±50.92)ng/ml,(215.95±131.26)ng/ml,(252.65±84.42)ng/ml,(337.13±60.52)ng/ml。The ATP concentration was at lowest level by the7d,then increasedslowly,but it was significiantly lower than of pretransplantation by the30days(p<0.05),especially in the antibody-inducting therapy group. There were13patients who occurredacute cellular rejection. The ATP concentration of those patients before transplantation,acute rejection,effectivetreatment of14d were (338.36±103.03)ng/ml,(507.02±125.37)ng/-ml,(372.53±92.73)ng/ml。The ATP values showed a significant increase when therejection occurs and decline after effective treatment. There were20patients who occurred lung infection. The ATP concentration of those patients before transplantation, infection,effective treatment of14d were (383.04±84.58)ng/ml,(138.74±55.57)ng/ml,(238.16±31.40)ng/ml.The ATP concentration discreased during lung infection andincreased after treatment.In addition,no direct correlation was observed between ATPconcentrations and therapeutic drug trough levels in whole biood(P>0.05).Conclusions:1.It is feasibility to monitor the cell immune function of renal transplant recipients byusing Cylex-immuknow technology.2.Using Cylex-ImmuknowTMtechnology moitoring intracellular ATP concertrationof CD4~+T-lymphocyte,.The ATP values showed a significant increase when the rejectionoccurs and discreased during lung infection,It may have some roles in the prediction ofrejection and lung infection.3.The early cellular immune function of the renal transplant recipients can be reducedby using basiliximab.4.Adjusment of the plama concentration can balance immunosuppressioninsufficiency and overdose.However, studies have shown that no direct correlationbetween the CSA/FK506plasma concentration and the concentration of theATP,further demonstrates the importance of a comprehensive assessment of thecell-mediated immunity function in the individual treatment.5.The use of Cylex-ImmuknowTMtechnology monitoring the cell-mediated immunefunction of the renal transplant recipients has the advantages of fast, sensitive, whice canadjust the immunosuppressant dosage,and optimize the treatment program.