The Measurement And Significance Of The HLA Class-Ⅰ (-A, -B, -C) Expression On Peripheral Blood CD3~+/CD8~+ T-Lymphocyte Surface After Renal Transplant

BACKGROUND:Accompany with the continual advancement of the immunosuppressive,renal transplant developed rapidly.The rates of acute rejection after transplant decreased,the long-term survival rates increased,renal transplant is the most effective therapy for end-stage renal disease(ESRD).But acute rejection(AR) is a major complication after renal transplantation and the biggest risk factor leads to chronic rejection.The rates of acute rejection were different in each transplant center,about 30%-50%.In a sense,the history of the renal transplant was a struggle procedure to the acute rejection.How to command the transplant recipients' immune status,administrate the immunosuppressive correctly,make a balance between the immune insufficient and immune excess, are still a puzzle for the doctors all over the world.Nowadays,the biopsy is remain the reliable diagnosis methods in acute rejection and is regarded as "golden standard",but it is invasive,it also has many potential complications, and the patients are unwilling to accept it,all of these restrict its clinical application.How to use a noninvasive or lower invasive methods to measure the acute rejection timely and accurately without notable damage of the allograft,is a hot spot in the current researches in the world.OBJECTIVE:AR have the negatively impact to the long-term survival of the allograft.No clinical markers of renal rejection have been universally accepted.In our previous study,we found the expression of HLA class-Ⅰon the peripheral blood surface,especially the expression of HLA-A,HLA-B could reflect the host immune status.And they encode non-polymorphism situs were decreased in many oncoma and kidney transplant patients,and the expression of HLA class-Ⅰincreased during the rejection process.This study was to explore if the expression of HLA class-Ⅰ(-A,-B,-C) on the peripheral blood CD3~+/CD8~+ T-lymphocytes could an indicator in predicting the AR after renal transplantation.METHODS:Seventy-nine renal transplant recipients were collected during the period of September 2007 to November 2008.Peripheral blood samples were obtained immediately before the operation.After the transplantation,blood samples were obtained at days 3 and 7 when the patients were still hospitalized, and followed at weeks 2,3 as well as months 1,2,3,and 6 after transplantation. For those patients with fever,lumbodynia,gross hematuria or oliguria,blood samples were collected immediately before the administration of anti-rejection regents and at days 3 and 7 after the administration.The levels of HLA class-Ⅰ(-A,-B,-C) on the peripheral blood CD3~+/CD8~+ T-lymphocytes were measured using flow cytometry.In the AR patients,Paraffin-embedded section of allograft kidney biopsies samples were sliced and immunostained using the anti-HLA-ABC antibody to measure the expression of HLA class-Ⅰon the lymphocytes.RESULTS:The levels of HLA class-Ⅰ(-A,-B,-C) on the peripheral blood CD3+/CD8+ T-lymphocyte were consistently elevated during the first three weeks and declined gradually to the levels before the transplantation,afterward tapered and kept stable.There were no significant changes on HLA class-Ⅰ(-A, -B,-C) levels among the patients during the routine checkout either before the operation or during the 6-month period after the operation,except two time points at 14 and 21 days after transplantation(P＜0.01).The AR occurred at the days 3,5,7,10,14,16,16,22,25,38,39,45,67.The levels of HLA class-Ⅰ(-A,-B,-C) increased significantly for all 13 patients during the acute rejection episodes(ARE),there were significant statistical difference between the acute rejection time and the last routine checkout before acute rejection(P＜0.01).Then the patients were administrated with anti-rejection drugs,and 3,7days later the HLA class-Ⅰ(-A,-B,-C) levels lowered significantly but still higher than that before the AR.All the patients with AR were confirmed by biopsy.And the allograft kidney biopsies samples were evidenced by buffy around the lymphocyte.CONCLUSION:This study suggests that the up-regulation of HLA class-Ⅰ(-A,-B,-C) on the peripheral blood CD3~+/CD8~+ T-lymphocyte can be used as an accurate and reliable indicator in predicting the AR after renal transplantation.