Distribution Of Peripheral Blood B Lymphocyte Cycle In Renal Transplant Recipients During Perioperative Period And Antibody-Mediated Acute Rejection

Purpose: To study the distribution characteristics of peripheral blood B-lymphocyte cycle during perioperative and antibody-mediated acute rejection?AMR?in renal transplant recipients,and to explore the clinical significance of peripheral blood B-lymphocyte cycle for early diagnosis of AMR.According to the screening criteria,28 patients who underwent allogeneic renal transplantation in our hospital from April 2017 to March 2018 were selected as study objects.The subjects were divided into immediate graft function?IGF?group and AMR group..There were 23 patients in the IGF group,including 14 males and 9 females.There were 15 cases with FK506+MMF+Pred anti-rejection regimen and 8 cases with Cs A+MMF+Pred anti-rejection regimen.There were 5 cases in the AMR group,including 3 males and 2 females.There were 3 cases with FK506+MMF+Pred anti-rejection regimen and 2 cases with CSA+MMF+Pred anti-rejection regimen.All subjects underwent morning fasting cubital venous blood?EDTA anticoagulation?4 ml in the morning,IGF group: 1st day before surgery,1st day,3th day,5th day,7th day,9th day,and 11 th day after surgery;AMR group: 1st day before surgery,1st day,3th day,5th day,7th day,9th day,and 11 th day after surgery?with blood collected every other day?until the reversal of rejection?alone blood was taken on the day of rejection?.Clinical indicators include serum creatinine,white blood cell count,absolute lymphocyte count,lymphocyte ratio,etc.The relationship between B lymphocyte cycle and clinical observations was explored by examining the proportion of B lymphocytes in different periods.Cell cycle assays: lymphocytes were separatedby gradient centrifugation,PI staining,CD19 antibody labeling,and flow cytometry were used to measure cell cycle data using Novoexpress software.Results: 1.The characteristics of B lymphocyte cycle distribution in the IGF group The proportion of B lymphocyte in G₀/G₁ stage before operation was?75.52+6.28?%,and increased rapidly to?80.88+7.47?% after first days,and the difference was significant?P<0.001?;first days to 11 days after operation were stable at the level of 80-85%,and there was no statistical difference between each time point.The proportion of B lymphocyte in S stage was?11.16+4.53?%,and decreased to?7.42+4.09?%?P<0.01?after operation?P<0.01?,and then decreased further on the third day after operation,and then tended to be stable,and then decreased again after the brief rise of ninth days after operation.The proportion of B lymphocyte in G₂/M stage before operation was?7.98+2.12?%,and decreased to?5.98+3.44?%?P<0.01?after first days after operation,and was stable between 4-6% after the operation on the 1st-11 th day.The blood creatinine level decreased rapidly in the first 3 days after the operation,and tended to be stable after one week.The white blood cell count increased significantly on the first,second day after the operation,and then gradually recovered the level of stability.The absolute value of lymphocytes and the proportion of lymphocytes decreased significantly at first days after the operation,and then gradually recovered.The proportion of B lymphocyte in S phase was positively linear with creatinine,lymphocyte absolute value and lymphocyte proportion?P<0.01?,but no correlation with white blood cell count,but the proportion of B lymphocyte in G₂/M stage was not correlated with creatinine,white cell count,lymphocyte absolute value and lymphocyte proportion.2.The characteristics of B lymphocyte cycle distribution in the AMR group The B lymphocyte cycle in AMR group had significant changes before and after operation?stable phase,AMR and AMR?.Compared with preoperative,postoperative stable period G₀/G₁ The proportion of B cells increased significantly?P<0.01?,while the proportion of B cells in S and G₂/M phase decreased significantly?P<0.01?,while the proportion of B cells at each period of AMR was not significantly different from that beforeoperation.However,when AMR was reversed,the ratio of B cells in S?P=0.021?and G₂/M?P=0.03?was significantly lower than that before the operation.Compared with the postoperative stable period,the proportion of B lymphocyte in G₀/G₁ phase of AMR decreased significantly?P<0.01?,while the proportion of S?P=0.015?and G₂/M?P<0.01?cells increased significantly?P<0.01?,but when AMR was reversed,there was no significant difference in the proportion of B cells at each period compared with the post operation stable period.However,once the AMR was reversed,the proportion of B cells in G₀/G₁ phase was significantly higher than that in the AMR period?P<0.01?;accordingly,the proportion of B cells in S?P=0.033?and G₂/M stages?P=0.028?was significantly lower than the period of AMR.The results showed that there were significant changes in all stages of B lymphocyte during the occurrence of AMR,and the level of B lymphocyte could be recovered to the stable stage after the rejection.The blood creatinine level of the recipients in group AMR showed a steady decline after the operation,but it increased significantly at the time of AMR,and then recovered to the stable stage after the reversal of AMR.The white blood cell count increased in the stable period and AMR after the operation,and then decreased after the reversion,but still higher than the preoperative level;the absolute value of lymphocytes and lymph nodes.The proportion of cells decreased significantly in the stable phase after operation,but increased at AMR,but it was still lower than the preoperative level.After AMR reversal,it decreased to a stable stage.There was no correlation between S phase of B lymphocyte in AMR group and serum creatinine,white blood cell count and lymphocyte proportion,but positive linear correlation with lymphocyte absolute value?P<0.05?;G₂/M phase was not related to white blood cell count,but was positively linear with serum creatinine,lymphocyte absolute value and lymphocyte proportion?P <0.05?.3.Comparison of B lymphocyte cycle in different stages of group AMR and group IGF after operation Compared with the postoperative stable stage?D11?in group IGF,there was no statistical difference in the proportion of B lymphocytes in the stable period and the reverse of AMR in group AMR,while the proportion of B lymphocyte in G₀/G₁ stageincreased significantly?P<0.01?,while the proportion of B lymphocytes in S?P<0.01?and G₂/M phase?P=0.043?decreased significantly.4.The change of B lymphocyte cycle was earlier than that of serum creatinine when AMR occurred.In group AMR,the proportion of B lymphocyte S and G₂/M cells in peripheral blood of 5 patients increased respectively at fifth day,Ninth day,Ninth day,eleventh day and thirteenth day after operation,while the corresponding serum creatinine levels increased respectively on fifth day,tenth day,eleventh day,Twelfth day and fifteenth day after operation,which suggested that the changes of B lymphoid cell cycle were earlier than creatinine.Conclusions: 1.the activation and proliferation of B lymphocytes in IGF group were significantly inhibited after operation.The S phase of B lymphocytes was positively correlated with the absolute value of lymphocytes,lymphocyte ratio and creatinine.2.when AMR occurred,the proliferation of B lymphocytes increased significantly.After AMR reversal,the activation and proliferation of B lymphocytes were inhibited again and recovered to the level of postoperatively stable stage.At the time of AMR,the S phase of B lymphocyte was positively correlated with the absolute value of lymphocyte,while the G₂/M phase was positively correlated with the absolute value of lymphocyte,the proportion of lymphocytes and the level of creatinine.