流式细胞微球捕获技术检测肾移植受者外周血细胞因子的应用

摘要

目的 应用流式细胞微球捕获技术检测肾移植受者外周血细胞因子,分析其在巴利昔单抗应用前后、肾移植手术前后以及移植物排斥反应发生时的含量变化及临床意义.方法 72例肾移植受者根据术后移植物功能状况分为2组:肾功能稳定组53例,急性排斥反应组19例.根据巴利昔单抗使用与否分为2组:巴利昔单抗使用组32例,巴利昔单抗未使用组40例.采集72例肾移植受者术前、术后、急性排斥反应发生时和30名健康供者的外周血,应用流式细胞微球捕获技术分析IFN-γ、TNF-α、IL-10、IL-5、IL-4、IL-2共6种细胞因子含量.对肾移植供者与受者组、肾移植受者术后功能稳定组与急性排斥反应组、术后4周巴利昔单抗使用组与未使用组、巴利昔单抗使用前后细胞因子含量进行比较.结果 术前肾移植受者组外周血TNF-α、IL-10、IL-5、IL-4、IL-2分别为(1.65±0.10)、(2.55±0.19)、(1.88±0.14)、(1.85±0.12)、(2.12±0.09)ng/L,低于健康供者组的(3.04±0.17)、(3.33±0.26)、(4.03±0.25)、(2.73±0.16)、(4.03±0.26)ng/L,差异有统计学意义(t值分别为6.890、2.375、7.851、3.955、7.153,P值分别为<0.01、<0.05、<0.01、<0.01、<0.01);术前肾移植受者组IFN-γ含量为(2.50 ±0.18)ng/L,健康供者组为(3.00±0.24)ng/L,差异无统计学意义(t=1.625,P>0.05).肾功能稳定组IFN-γ、IL-10分别为(2.71±0.11)、(3.91±0.52)ng/L,低于急性排斥反应组的(3.30±0.36)、(12.01±5.35)ng/L,差异有统计学意义(t值分别为5.061、11.465,P值分别为<0.01、<0.05).使用巴利昔单抗的肾移植受者,在巴利昔单抗使用后IFN-γ、TNF-α和IL-10分别为(2.78±0.17)、(1.58±0.07)、(2.77±0.24)ng/L,与使用前的(2.90±0.21)、(1.67±0.12)、(2.45±0.16)ng/L比较,差异有统计学意义(t值分别为5.605、6.011、4.126,P值分别为<0.01、<0.01、<0.05).术后4周巴利昔单抗使用组IFN-γ、IL-10、IL-4分别为(2.90±0.31)、(9.08±0.16)、(2.73±0.11)ng/L,与未使用组的(3.28±0.11)、(4.17±0.21)、(2.11±0.20)ng/L比较,差异有统计学意义(t值分别为4.268、4.263、3.762,P值分别为<0.01、<0.01、<0.05).结论 流式细胞微球捕获技术使用微量标本可同时检测6种细胞因子.联合检测肾移植受者外周血中的细胞因子能够为临床诊疗提供更有意义的指导依据.%Objective To measure the cytokines levels in peripheral blood from kidney transplantation recipients by using cytometric bead array and to analyze their change and the clinical significance in pre- and post- kidney transplantation, inducting with basiliximab and graft rejection. Methods A total of 72 renal transplantation recipients were divided into two groups, kidney function stable group(n =53) and acute rejection group (n = 19). And they were also grouped by induction with basiliximab or not,32 in basiliximab group and 40 in without basilixmab group. The levels of IFN-γ, TNF-α, IL-10, IL-5,IL-4, IL-2 were measured by cytometric bead array in peripheral blood of 72 kidney transplantation recipients and 30 healthy donors at differential time. The data was analyzed according to the following grouping:donors and recipients, kidney function stable group and acute rejection group post transplantation and with or without basiliximab group. Results The levels of TNF-α, IL-10, IL-5, IL-4, IL-2 in recipients before transplantation were ( 1.65 ±0. 10) ,(2. 55 ±0. 19) ,( 1.88 ±0. 14) ,(1.85 ±0. 12) ,(2. 12 ±0. 09) ng/L,respectively. While they were (3.04 ±0. 17), (3.33 ±0. 26), (4.03 ±0.25), (2.73 ±0. 16), (4.03 ±0. 26) ng/L respectively in healthy donors. There was statistical significance between the two groups ( t =6. 890, 2. 375, 7. 851,3.955,7.153, P<0. 01, <0. 05, <0.01, <0.01, <0.01). While the level of IFN-γ in recipients before transplantation was (2. 50 ±0. 18) ng/L,compared with (3. 00 ±0. 24) ng/L in healthy donors. There was no statistical significance between the two groups( t = 1. 625, P > 0. 05 ). The levels of IFN-γ and IL-10 in kidney function stable group were (2. 71 ± 0. 11 ) ng/L and (3.91 ± 0. 52) ng/L,while they were ( 3.30 ± 0. 36 ) ng/L and ( 12. 01 ± 5.35 ) ng/L in acute rejection group. There were statistical dirrerences between the two groups ( t = 5. 061, 11. 465, P < 0. 01, < 0. 05 ). Before induction with basiliximab, the levels of IFN-γ, TNF-α, IL-10 in recipients were (2.90 ±0. 21 ), ( 1.67 ±0. 12),(2. 45 ± 0. 16) ng/L respectively. But they were ( 2. 78 ± 0. 17 ), ( 1.58 ± 0. 07 ), ( 2. 77 ± 0. 24 ) ng/L respectively after induction with basiliximab, which showed significantly different ( t = 5. 605, 6.011,4. 126, P <0. 01, <0. 01, <0. 05). Four weeks after kidney transplantation in recipients with basiliximab,the levels of IFN-γ, IL-10, IL-4 were (2. 90 ± 0. 31 ), (9. 08 ± 0. 16), (2. 73 ± 0. 11 ) ng/L. While they were (3.28 ±0. 11 ), (4. 17 ±0. 21 ), (2. 11 ±0. 20) ng/L respectively in recipients without basiliximab induction, which were significantly different from those with basiliximab induction (t = 4. 268,4. 263,3.762, P <0. 01, <0. 01, < 0. 05 ). Conclusions Six kinds of cytokines can be measured by cytometric bead array simultaneously and accurately. The data suggests that the detection of multiple cytokines in kidney transplantation recipients by cytometric bead array can provide more guidance for clinical diagnosis and therapy.