| The effective treatment for end-stage renal disease is kidney transplantation. With the continuous updating and development of immunosuppressive drugs, the probability of occurrence of acute renal allograft rejection gradually reduced and graft survival has been greatly improved.But all along,because there is no a accurate indicator for monitoring immune function after kidney transplantation,how to objectively and accurately assess the immune status of patients after transplantation,and to strike a balance between excessive immunosuppression and inadequate immunosuppression,then to guide the individual drug usage has become an important issue for transplant doctors to urgently address and research.Nowadays,thetransplant surgeon often through invasive and non-invasive means to monitor immune status after renal transplantation,including the general condition of the receptor, serum creatinine, color Doppler ultrasound, monitor immunosuppressant drug concentration in blood, graft biopsy histopathology etc. But sensitivity and specificity of serum creatinine is poor,and it is little help for clinical prediction and treatment. The ultrasound diagnosis has some subjective effects by a great doctor.Now, the commonly means which is used in clinical to assess the immune status is monitoring of serum concentrations of immunosuppressants. However, because the drug metabolism in different recipients is different,and a variety of factors in the body can affect the immune serum concentration of drug,so the acute rejection is inevitablly happened in vivo.The graft biopsy is an invasive examination,and with a certain risk for patients,so it is difficult to repeatedly monitor in clinical.Base on the above research works,we try to detect the proportions of T lymphocyte and their subsets in peripheral blood of renal transplant recipients in order to monitor their immune states. We endeavor to clarify the immune responses characteristics of renal transplant recipients by comparing the percentages of T lymphocyte and its subsets and evaluating balance in healthy donors, stable renal function recipients and uremic patients.Meanwhile, we dynamically observed the changes of T lymphocyte subsets percentages in the peripheral blood in stable renal function recipients, acute rejection recipients,infection recipients, explored the roles of T cells and its subsets in renal transplant recipients. Finally, we try to compare the proportions of the acute rejection and infection recipients in the patient did not use T cells,T cell monitoring group, then to verify the dynamic monitoring of T cells is contribute to detect the immune status of transplantation. This may provide a new criterion for the individual drug usage.Part I The different proportion of the percentages of T lymphocyte and its subsets and evaluating balance in healthy donors, stable renal function recipients and uremic patients.ObjectiveWe try to clarify the immune responses characteristics of renal transplant recipients by comparing the percentages of T lymphocyte and its subsets and evaluating balance in healthy donors, stable renal function recipients and uremic patients,and combined with the recipient’s status observation before renal transplantation and T cell changes after operation, in order to detect variation of T lymphocyte between before renal transplantation and after transplantation.MethodsBy flow cytometry,we try to detect the proportions of T lymphocyte and their subsets in peripheral blood of renal transplant recipients in order to monitor their immune states. We endeavor to clarify the immune responsescharacteristics of renal transplant recipients by comparing the percentages of T lymphocyte and its subsets and evaluating balance in25healthy donors,25stable renal function recipients and25uremic patients.Results①The percentage of CD3+T lymphocytes have no statistical significance between in the healthy group, uremia group and normal group (P>0.05).The percentage of CD3+CD4+T lymphocytes was no significant difference between the healthy group and uremia group (P>0.05), and the two groups were compared with normal renal function group, the differences were statistically significant (P<0.05). The percentage of CD3+CD8+T lymphocytes in healthy group, uremic group and the normal group was not statistically significant (P>0.05). CD3+CD4+/CD3+CD8+was no significant difference (P>0.05) between the three groups.②The count of CD3+CD4+T lymphocytes was no significant between the health group and the uremia group (P>0.05).while the two group were compared with the stable group, the difference was statistically significant(P<0.05). CD3+CD8+T lymphocyte count was no significant between the healthy group, the postoperative uremia group and stable group (P>0.05).ConclusionFirst, Immunosuppressive drugs have effects on the count of CD4+T cells. Second, serum creatinine levels, peritoneal dialysis, hemodialysis and blood transfusions and other medical-related treatment has little effect on the proportions of T lymphocyte and their subsets.Part Ⅱ Comparing the percentages of T lymphocyte and its subsets in different renal transplantation recipientsObjectiveBy flow cytometry,we try to detect the proportions of T lymphocyte and their subsets in peripheral blood of renal transplant recipients in order to monitor their immune states. We endeavor to clarify the immune responses characteristics of renal transplant recipients by comparing the percentages of T lymphocyte and its recipients.MethodIncluded69cases of kidney transplant patients,who received allogeneic cadaveric donor kidney and relatives living donor renal transplant patients. The serum creatinine all the selected patients decreased to normal levels within two weeks after surgery. Those patients did not had occured hyperacute rejection in surgery, and without anyr diseases occur during testing, which affecting the immune status of the patient. According to the condition of the patient,we divided into casesinto three different groups:the normal renal function group (n=25), acute rejection group (n=23) and lung infection group (n=21). Those recipients have the same blood group ABO with donor, blood lymphocyte toxicity test<10%,PRA<15%, the number of HLA mismatches were less than4. By flow cytometry,we try to detect the proportions of T lymphocyte and their subsets in peripheral blood of renal transplant recipients by comparing the percentages of T lymphocyte and its subsets in stable renal function recipients, acute rejection recipients,infection recipients.The resulting data using SPSS13.0software for analysis. All of the data were obtained through the normality test and homogeneity of variance test, mean±standard deviation (mean±SD) said.Measurement data between the two groups were compared using t test or rank sum test; multiple groups were compared using ANOVA, P<0.05was considered statistically significant.Results①The data in three groups has no significant difference in gender, age and constitute the main immunosuppressant (P>0.05).②The percentage of CD3+T lymphocytes in the three groups was not statistically significant (P>0.05). The percentage of CD3+CD4+T lymphocytes in acute rejection group is highest, compared with normal renal function after transplantation group, the differences were statistically significant (P<0.05). While the percentage of CD3+CD4+T lymphocytes in the infected group is lowest than other groups, compared with normal renal function after transplantation group, the difference was statistically significant (P<0.05). The percentage of CD3+CD8+T lymphocytes in acute rejection group was the lowest, compared with normal renal function after transplantation group, the difference was statistically significant (P <0.05). While the percentage of CD3+CD8+T lymphocytes was the highest in the three groups, compared with the group with normal renal function after transplantation, the differences were statistically significant (P<0.05).The ratio of CD3+CD4+/CD3+CD8+in acute rejection group was highest than other groups, while the ratio of CD3+CD4+/CD3+CD8+in the infected group is lowest, both groups compared with normal renal function after transplantation group, the difference was statistically significant (P<0.05,P<0.05).The count of CD3+CD4+T lymphocytes in acute rejection group was highest than others/while compared with normal renal function after transplantation,and the difference had statistical significance (P <0.05). While CD3+CD4+T lymphocytes count in infected group was the lowest in the three group,while compared with normal renal function after transplantation,and the difference had statistical significance too(P<0.05).The count of CD3+CD4+T lymphocytes in acute rejection group was compared with other groups,the difference had statistical significance(P<0.05,P<0.05).③The percentage of CD3+T lymphocytes in the infected group and after treatment has was not statistically significant difference (P>0.05).After the positive treatment, the percentage of CD3+CD4+increased from19.87±7.95to38.42±7.69, the difference wasstatistically significant(P<0.05); and the ratios of CD3+CD4+/CD3+CD8+ratios also increased from0.36±0.74to1.25±0.68,the difference was statistically significant too (P<0.05).While the percentage of CD3+CD8+decreased from46.76±8.58to35.28±7.15, the difference was statistically significant (P<0.05). The count of CD3+CD4+T lymphocytes increased from208±151to462±179, and the difference was statistically significant(P<0.05). The count of CD8+T cell was reduced from316±182to393±187, and the difference was statistically significant(P<0.05).④The percentage of CD3+Tlymphocytes inthe acute rejection group than after treatment has was not statistically significant difference (P>0.05). After the positive treatment,the percentage of CD3+CD4+decreased from49.14±7.81to38.26±7.25, the difference was statistically significant (P<0.05); and the ratio of CD3+CD4+/CD3+CD8+reduced from2.53±0.68to1.45±0.73, and the difference was statistically significant (P<0.05).While the percentage of CD3+CD8+increased, from21.37±8.26to39.83±8.15,the difference was statistically significant (P<0.05).The count of CD4+T cell was reduced from697±189to546±205,and the difference was statistically significant (P<0.05).The count of CD8+T cell was reduced from363±194to418±216,and the difference was statistically significant(P<0.05).Conclusion①CD3+CD4+T lymphocyte is closely related with the graft rejection and infection.②If we observed the changes of T lymphocyte subsets by dynamic, especially to observe the changes of CD4+T cell,and it wll be helpful for the clinical medication.PartⅢComparing the proportions of the complications between T cell monitoring of renal transplant recipients and the control group of recipientsObjectivewe try to detect the proportions of T lymphocyte and their subsets in peripheral blood of renal transplant recipients in order to monitor their immune states. According to the change, the immunosuppressant drugsdosage could be tweaked, and it wll be helpful for the clinical medication.Method This reasch included52patients,who received kidney transplantation because of the chronic renal failure. According to whether or not monitor Tlymphocytein peri operation period and after June,we divided the case into T cell monitoring group and did not use T cell monitoring group. A detailed record of information of the two groups of patients, including: Gender, age, weight, donor type, matches, immune induction therapy, based immune suppression scheme, serum creatinine (Scr), delayed graft function (DGF), acute rejection (AR), infection, clinical data/kidney survival,etc. We detect CD3+CD4+T cell count one per week in preoperative and postoperative3months, and according to the level of CD3CD4+T cell count adjust immune induction and immune inhibitor,then the target of CD3+, CD4+T cell count would be controlled in300-500/uL or preoperative level of60%to80%according to the previous references and our experience.And we endeavor to observe the probability of acute rejection and infection,and compare the probability in two group of renal transplant recipients.Results①The data in three groups has no significant difference in Gender, age, weight and HLA match, donor type, basic protocols, antibody induction therapy and the incidence of DGF (P>0.05).②The mean of creatinine in monitoring group1month after renal transplantation is140.8±22.0μmol/L, while it is145.6±37.6μmol/Lin6month after renal transplantation. The mean of creatinine in the control group1month after renal transplantation is153.5.4±25.5μmol/L, while it is149.2±59.1μmol/Lin6month after renal transplantation. It has no significant difference between the two groups.③The incidence of acute rejection in the monitoring group was lower than the control group,while the incidence of infection has no significant difference between the two groups.ConclusionIf we observed the changes of T lymphocyte subsets by dynamic, it wll be helpful for the clinical medication. |
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