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目的研究ELISA检测肾移植受者外周血IL-2水平与实时定量PCR(RT-FQ-PCR)检测外周血淋巴细胞(PBL)IL-2 mRNA表达结果的一致性和准确性。方法对20例肾移植受者,采用ELISA和RT-FQ-PCR法分别测定术前、术后7天谷浓度(C0)和峰浓度(C2)时间外周血IL-2和PBLIL-2 mRNA水平;同时选取15例慢性移植肾肾病(CAN)和5例非CAN患者,测定方法同上。结果外周血IL-2水平与PBLIL-2 mRNA表达在术前和手术1年后呈正相关(R2分别为0.58和0.40,P均<0.01),在术后7天C0及C2时间无明显线性关系(R2分别为0.15和0.08,P均>0.05)。术后7天,C2对应外周血IL-2和PBLIL-2 mRNA表达抑制率分别为16.31%±9.59%和69.92%±7.10%;C2与IL-2 mRNA表达抑制率呈正相关,与IL-2抑制率无明显相关性(R2=0.02,P>0.05)。IL-2 mRNA表达抑制率与IL-2抑制率无明显相关性。结论外周血IL-2ELISA检测可以用于肾移植受者术前和术后稳定期免疫状态检测以及免疫耐受研究等,但不能用于肾移植受者围手术期免疫状态和C2效应检测。
Objective To study the consistency and accuracy of ELISA for the detection of IL-2 mRNA in peripheral blood from kidney transplant recipients and the detection of IL-2 mRNA expression in peripheral blood lymphocytes (PBL) by real-time quantitative polymerase chain reaction (RT-FQ-PCR). Methods Twenty (20) renal transplant recipients were enrolled in this study. The levels of IL-2 and PBLIL-2 mRNA in peripheral blood were measured by ELISA and RT-FQ-PCR at preoperative and postoperative 7 days, respectively. At the same time, 15 cases of chronic allograft nephropathy (CAN) and 5 cases of non-CAN were selected. Results The level of IL-2 in peripheral blood was positively correlated with the level of PBLIL-2 mRNA 1 and 2 years after operation (R2 = 0.58 and 0.40, P <0.01 respectively), but there was no significant linear relationship between C0 and C2 at 7 days after operation (R2 0.15 and 0.08, P> 0.05 respectively). After 7 days, the inhibitory rates of IL-2 and PBLIL-2 mRNA in C2-corresponding peripheral blood were 16.31% ± 9.59% and 69.92% ± 7.10% respectively. The inhibitory rates of C2 and IL-2 mRNA were positively correlated with IL- No significant correlation between inhibition rate (R2 = 0.02, P> 0.05). IL-2 mRNA expression inhibition rate and IL-2 inhibition rate was not significantly correlated. Conclusion The detection of IL-2 ELISA in peripheral blood can be used to study the immune status and immune tolerance in preoperative and postoperative stable renal transplant recipients, but can not be used in perioperative immune status and C2 effect in renal transplant recipients.