肾移植受者外周血IL-2 ELISA检测与PBLIL-2 mRNA实时定量PCR检测的比较

Comparative study of peripheral blood IL-2 determination by ELISA with PBL IL-2 mRNA expression by real time PCR in recipients of renal transplantation

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 compare the consistency and accuracy of determination of peripheral blood lymphocyte （PBL） interleukin- 2 （IL-2） with ELISA and evaluation of the expression of PBL IL-2 mRNA with real time PCR（RT-PCR）. Methods In 20 kidney recipients, the PBL IL-2 level was determined with ELISA and the mRNA expression in PBL IL-2 was evaluated by real time fluorescence quan- titative PCR （RT-FQ-PCR） before operation and C0/C2 time on the 7th day after operation. The same detection and evaluation were also done in 15 chronic allograft nephropathy （CAN） patients and 5 non CAN patients. Results IL-2 and IL-2 mRNA expression showed positive correlation before operation and 1 year later after operation, R^2=0. 58 and 0. 4 respectively, both P〈0. 01. No apparent linear relation was found at CO and C2 time on the 7th day after operation, Re =0. 15 and 0. 08 respectively, both P〉0. 05. On the 7th day after operation, IL-2 and the inhibition rate of IL-2 mRNA expression were （16. 31±9. 59）% and （69. 92±7. 10）%, respectively. A positive correlation existed between C2 IL-2 concentration and the inhibition rate of IL-2 mRNA expression, Re =0. 56, P〈0. 01, while no apparent correlation was found between （22 IL-2 concentration and the IL-2 inhibition rate, Re =0. 02, P〉0. 05. Also, IL-2 inhibition and PBL IL-2 mRNA inhibition showed no linear correlation. Conclusions The technique of determining PBL IL-2 by ELISA can be used for evaluating immunity status and immunosuppressive tolerance in patients with renal transplantation before the operation and at stable stage after operation. However, it should not be used for assessing perioperative immunity status and evaluating effect of cyclosporine.