A Study On The Change Of T Lymphocyte Subgroups In The Peripheral Blood Of Long-Term Surviving Patients Who Have Received Liver Transplantations

The liver transplantation has already become an effective method to treat terminalstage liver diseases. Rejection reactions of liver transplantations are easy to control,compared with that of other organs transplantations. In liver-kidney, liver-pancreas, andother multiple organ transplantations, the liver transplantation can effectively lead theacceptors to produce immune tolerance to the kidney and the pancreas, and consequentlyother organs can be protected to some extent[1]. In recent years, helper T cells andsuppressor T cells have been paid more and more attention in immune responses after theliver transplantation. T cells play a very important part in taking part in rejection reactionsof transplantations and inducing the immune tolerance. By means of a large number ofanimal experiments and clinical tests, the functional mechanism and characteristics of somelymphocytes and cytokines in the transplantation immunity have been clarified and hascome into use in the clinical practice.After the liver transplantation, the change of patients’ immunological function ismainly reflected in the change of T lymphocyte subgroups and cytokines levels. The levelof different subgroups and their cytokines varies in the rejection reactions and theperipheral blood of immune tolerance producers. The eventual balance of the level is aguarantee for patients to produce persistent immune tolerance. The regulatory T cell （helperT cell and suppressor T cell） is the key factor to maintain the balance between the rejectionreaction and the immune tolerance, which is decided by the balance between the effector Tcell and the regulatory T cell. The restriction of the effector T cell function and thereduction of patients’ rejection reaction after the liver transplantation can be only achievedby activating the inhibiting effect of the regulatory T cell.Existing researches have shown that the regulatory T cell can exert modulatory effect on a variety of immune cells[2-4]. Animal experiments have confirmed that the regulatory Tcell plays an important role in significantly reducing and relieving graft-versus-hostdiseases[5-7]. Lots of scholars have made some progress in applying CD4⁺CD25⁺T cellsexpanded in vitro to reduce graft-versus-host diseases after the mice receive thetransplantation[8-10]. The author’s department has made the peripheral blood of long-termsurviving patients having produced immune tolerance after liver transplantations the targetof the research, tested levels of Th1cells, CD3⁺CD8^lowsuppressor T cells（Ts）, andCD4⁺CD25⁺Foxp3+Treg, discussed the change of Th1cells, CD3⁺CD8^lowsuppressor Tcells（Ts）, and CD4⁺CD25⁺Foxp3+Treg in the peripheral blood of patients who haveproduced immune tolerance after liver transplantations, and provided a indicative basis forthe immune tolerance production and drug use of patients who have received livertransplantations.ObjectiveTo analyse the change of Th1cells、 CD3⁺CD8^low suppressor T cells andCD4⁺CD25⁺Foxp3+Treg level in the peripheral blood of the long term survival patientunderwent liver transplantation, and to determine its significance in clinical laboratory andimmunology.Method1. The peripheral blood of20healthy people is used as the control group. Theperipheral blood of20patients who have survived for2to3years and20patients who havesurvived for8to10years is taken, and the heparin is used as anticoagulant. The expressionof AST,ALT,GGT and FK506in the serum was detected by serological analysis.2. The PBMC cells are separated, PMA is used to stimulate the proliferation, and thecells are collected. Three groups A, B, C are divided.10ulAnti-IL-2/FE is added to group A,10ul Anti-CD3/PE and10ul Anti-CD8/FITC are added to group B, and10ul Anti-CD4/PerCP-cy5.5,10ul Anti-CD25/FE, and10ul FoxP3McAb/APC are added to group C.Cultivate the cells away from light under4℃for1hour, put them in PBS resuspensionsolution, centrifuge them for3minutes at3000rpm, wash them twice. Put the cells in150ulPBS resuspension solution and test them with flow-cytometer machine. 3. Statistical analysis: Statistical software SPSS13.0is used to analyze the data,x±sis used to present the measurement data, which is processed by variance analysis and t-test,and the difference is significant when p<0.05. The Pearson method is applied to analyze therelevance, there is relevance when p<0.05.Result1. There was not a significant difference of the expression of AST, ALT,GGT andALB in the peripheral blood of patients with liver transplantation for2to3years, patientswith liver transplantation for8to10years, and healthy people, but the level of GGT in theperipheral blood of patients with liver transplantation was increased significantly thanhealthy people. FK506was7.58±0.47ng/ml in the peripheral blood of patients with livertransplantation for2to3years, and4.21±0.3ng/ml in the peripheral blood of patients withliver transplantation for8to10years. The expression of FK506was overexpressedsignificantly in patients with liver transplantation than in healthy people, but Theexpression of FK506in patients with liver transplantation for8-10years was decreasedsignificantly than in patients having survived for2-3years.2. The radiation of Th1cells, CD3⁺CD8^lowsuppressor T cells（Ts）andCD4⁺CD25⁺Foxp3+Treg expression was32.15±3.05%,23.34±2.87%,17.87±3.22%respectively in the peripheral blood of patients with liver transplantation for2to3years,and18.71±2.97%,18.32±2.54%,10.76±2.21%respectively in the peripheral blood ofpatients with liver transplantation for8to10years, and12.26±2.35%,10.74±3.21%，6.43±1.12%respectively in healthy people. The levels of Th1cells, CD3⁺CD8^lowsuppressor T cells（Ts）and CD4⁺CD25⁺Foxp3+Treg was overexpressed significantly inpatients with liver transplantation for2to3years than in healthy people, and only theexpression of Ts in patients with liver transplantation for2to3years was significantlyincreased than in healthy people. Furthermore， the expression of Th1cells andCD4⁺CD25⁺Foxp3+Treg in patients with liver transplantation for8-10years wasdecreased significantly than in patients with liver transplantation for2-3years. The threelevels eventually are close to levels of healthy people.3. With multiple linear regression to analyze the correlation of FK506and Th1cells, CD3⁺CD8^lowsuppressor T cells（Ts）and CD4⁺CD25⁺Foxp3+Treg after liver transplantation,we found that the expression of FK506intimately correlate with the levels of of Th1cells,CD3⁺CD8^lowsuppressor T cells（Ts）and CD4⁺CD25⁺Foxp3+Treg.Conclusion1. Immune tolerance production of patients after liver transplantation is to some degreedecided by the balance of Th1cells, CD3⁺CD8^lowsuppressor T cells（Ts）, andCD4⁺CD25⁺Foxp3+Treg.2. The content changes of Th1cells, CD3⁺CD8^lowsuppressor T cells（Ts）, andCD4⁺CD25⁺Foxp3+Treg in the peripheral blood of patients after liver transplantation maybe the drug use indicator compared with biochemical indicator to some degree.