| PURPOSE Study immunity status and CsA effect of early KT and CAN patients, explore the relation of TGF-pl with CAN, compare the coincidence and accuracy of measuring peripheral blood (PL) interleukin-2 by ELISA with evaluating PBLIL-2 mRNA expression by real time PCR.MATERIAL AND METHODS By real time fluoresence quatitative polymerase chain reaction(RT-FQ-PCR), we tested some cytokines mRNA expression in PBL of 32 KT patients preoperation and 3, 7, 14, 28 days later after operation;For 1 case of DGF and 2 cases of AR patients, added measurements after DGF or AR happened. For 20 cases of patients who used CsA use, added C2 time test and measured CsA concentration at 0 and 2 hour 7 days later after operation;measured peripheral blood IL-2 level by ELISA preoperation and CO and C2 time of 7 days later after operation. Took 10 cases of patients who applied for routine operation as controls, tested cytokines mRNA expression preoperation and 3, 7 days later after operation. For 15 cases of CAN and 22 cases of non CAN patients, grouped them according to CsA or MMF use, measured some cytokines mRNA expression in PBL;for 20 of which, measured peripheral blood IL-2 level by ELISA.RESULTS There was no difference in cytokines mRNA expression between KT and OC group before operation;IL-10 mRNA expression of KT group was higher than that of OC group after operation, but IL-2, IFN-γ mRNA expression was apparently lower than that of OC group 7 days later after operation. For 2 cases of early AR patients, IL-2. IL-10. IFN-γ, TGF-β1 mRNA expression got stronger before and aftert AR happening, high IL-2 mRNA expression appeared most apparently. 7 days later after operation, IL-2, IFN-γ mRNA inhibited rate of C2 time was (69.47±7.09)% and (65.95±8.15)% respectively;it had fine correalation with C2 concentration, R~2=0.53 and 0.31 respectively, P<0.01 and <0.05 respectively;it had no correlation with COconcentration. IL-2 and IL-10 mRNA expression of CAN group was lower than that of non CAN group, IFN-y and TGF-pl mRNA expression was higher than that of non CAN group apparently, P<0.01. TGF-pi mRNA expression of CsA group was higher than that of FK506 group, P<0.01;TGF-Pl mRNA expression appeared negative correlation with allograft function, P<0.01. Peripheral blood IL-2 and PBL IL-2 mRNA expression showed positive correlation before operation and during 1—5 year after operation, R2=0.58 and 0.39 respectively, both P <0.01;they appeared no correlation at CO and C2 time 7 days later after operation, R2=0.16 and 0.08 respectively, both P >0.05. 7 days later after operation, the inhibited rate of peripheral blood IL-2 was lower apparently than that of PBL IL-2 mRNA expression, P<0.01;C2 Concentration and IL-2 inhibited rate appeared no fine correlation, R2=0.05, P >0.05. IL-2 inhibited rate and PBL IL-2 mRNA inhibited rate had no correlation also, R2=0.09, P>0.05.CONCLUSIONS Measuring cytokines mRNA expression in PML by RT-FQ-PCR may be an effective method for detecting immunity status of KT patients and evaluating CsA effect. Thl and Th2 cytokines may both play critical role in the development and progression of AR and CAN, but maybe their mechanism is different at some extent;we still couldn't decide the development and progression of AR and CAN simply according to whether Thl/Th2 cell subset swift or not. Excessive depression of IL-2 may inhibit the function of T regulatory cell ( Tr), and then provoke the CAN development. CsA may cause high TGF-Pl mRNA expression, it may accompany with the development and progression of CAN, and affect graft function. IL-2 may coincide with IL-2 mRNA expression in patients with renal transplantation preoperation and at stable stage after operation, may lack of correlation perioperation. For KT patients, IL-2 measurement by ELISA may be considered for studying immunity status and immunotolerance preoperation and at stable stage after operation;but may not be suitable for measuring immunity satus of KT patients at early time after operation and evaluating cyclosporine effect perioperation.
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