免疫细胞功能分析在肾移植受者免疫状态监测中的应用

发布时间：2018-03-29 15:32

本文选题：肾移植　切入点：细胞免疫　出处：《浙江大学》2011年硕士论文

【摘要】：背景作为慢性肾功能衰竭患者最理想的肾脏替代疗法,肾移植开展半个多世纪以来,移植技术已基本攻克,但是排斥反应仍然是移植医生和病人面临的最大问题,也是移植物长期存活的最大障碍。器官移植受者在术后需要终生服用免疫抑制剂来避免排斥的发生,但是移植物存活率改善的同时受者发生药物中毒、机会性感染和癌症的风险也增加,而且基础免疫反应、手术创伤、麻醉、药物代谢动力学的个体差异等因素均可影响到免疫状态,所以必须进行移植术后免疫状态的监测。如何了解移植受者术后的免疫状态,合理应用免疫抑制药,在免疫抑制不足和免疫抑制过度之间取得平衡,是移植临床面临的重要课题。Cylex ImmuKnow法是唯一得到FDA认可的检测移植受者应用免疫抑制剂时细胞免疫(CMI)功能的方法。它直接反应细胞免疫的功能,针对免疫抑制用药环节,24小时之内可出结果,而且结果量化,是目前检测移植术后病人的免疫状态,指导用药的较好指标。但是,作为一项新的免疫学评价指标,ImmuKnow;法尚需在大范围、大样本量中验证与完善,尚需验证其是否符合中国人群的种族遗传背景。目的利用免疫细胞功能分析(ImmuKnow ATP测定)动态监测肾移植受者的细胞免疫反应性,分析免疫细胞功能与急性排斥,急性肾小管坏死和移植后感染的相关性,评估其在肾移植免疫状态监测中的价值。方法：健康成人20例；2010年3月-2010年12月在浙江大学医学院附属第一医院肾脏病中心首次接受同种肾移植受者,分别在术前、术后2w、lm、3m、6m及临床出现异常(排斥,感染等时间点)动态监测免疫细胞功能,同时纳入同期间出现排斥反应,感染等移植受者,共131例,根据临床情况分组：健康对照组(HC),肾移植术前组(Pre-Tx),术后稳定组(Tx),术后感染组(Infection),排斥反应组(AR),和急性肾损伤组,术后稳定、排斥和急性肾损伤组均经肾组织穿刺活检证实,感染组均经胸片、胸部CT或病原学检查证实诊断。抽取外周肝素钠抗凝血(2m1),进行细胞免疫功能分析(ImmuKnow ATP),同时采血2ml用于淋巴细胞亚群分析(CD4/CD8),1ml用于Fk506血药浓度检测,5ml用于混合淋巴细胞培养(MLR)。实验数据应用SPSS 16.0软件进行分析,两组之间比较采用T检验,排斥组与其他各组相比较采用One-Way ANONA, Hochberg's GT2法,P0.05有统计学意义,相关性分析采用Logistic回归分析方法。结果健康人ATP浓度为401±153ng/ml,肾移植受者术前ATP浓度405±161ng/ml,两者相比(中位数),肾移植术前组(317 ng/ml)ATP浓度低于HC组(404ng/ml)。ImmuKnow ATP浓度显示肾移植术后受者免疫状态出现波动,术后1个月时ATP浓度(603±76ng/ml)高于术后3个月(424±191 ng/ml)6个月(396±175 ng/m1)随访值,随着时间延长趋于平稳。AR组ATP浓度(539±217 ng/ml)与其他各组(Tx组332±155 ng/ml, Infection组288±140 ng/ml,AKI组336±125 ng/ml)比较均高于其他各组,差异有统计学意义(P0.05)。将移植受者的ATP浓度与MLR、CD4/CD8和Fk506血药浓度分别做相关性分析,发现ATP浓度与三者无明显相关,相关系数分别为R2=0.0072,R2=2E-06,R2=0.004。结论1本研究用最新免疫状态监测方法细胞免疫功能分析(ImmuKnow ATP浓度检测法)动态观察肾移植受者术后细胞免疫状态的变化规律,术后一个月内受者免疫状态处于较强状态,随着时间延长趋于稳定。2 ImmuKnow法检测的ATP值在急性排斥反应发生时有上升的趋势,对急性排斥反应的发生有一定的预示作用。3ImmuKnow ATP值可直接反应细胞免疫状态,与临床常用的检测方法CD4/CD8,MLR,Fk506血药浓度均无相关性。
[Abstract]:The background is the most ideal in patients with chronic renal failure renal replacement therapy, renal transplantation has carried out more than half a century, transplantation technology has been basically overcome, but rejection is still the biggest problem facing transplantation for doctors and patients, is the main obstacle of long-term graft survival. Organ transplant recipients require lifelong immunosuppression to prevent rejection the occurrence of postoperative graft survival, but at the same time improve the recipients of drug poisoning, the risk of opportunistic infections and cancer also increased, and the reaction of immune, surgical trauma, anesthesia, dynamic factors of individual differences in drug metabolism can affect the immune state, so it is necessary to monitor immune status after transplantation how to understand the immune state of transplantation. After the operation, the reasonable application of immunosuppressive drugs, in immunosuppression insufficiency and strike a balance between excessive, is An important issue facing the.Cylex ImmuKnow transplantation is the only FDA approved by the detection of transplantation application of cell-mediated immunity (CMI) function method. It cell immunity function in immunosuppressive drugs link, within 24 hours can be obtained, and quantitative results, is the detection of post transplant patients the immune state, better indicators to guide medication. However, as a new index, immunological evaluation of ImmuKnow; method is needed in a wide range, large amount of verification and improvement, still need to verify their compliance with the ethnic background China population.
Objective to analyze the function of immune cells (ImmuKnow ATP assay) dynamic monitoring of renal transplant cells immunoreactive to the analysis of immune cell function and acute rejection, acute tubular necrosis and infection after transplantation, evaluate the immune status of kidney transplantation in the monitoring value.
Methods: 20 healthy adults; March 2010 -2010 year in December for the first time of allogeneic renal transplant recipients at the Zhejiang University School of medicine, First Affiliated Hospital, respectively before surgery, postoperative 2W, LM, 3M, 6m and clinical abnormalities (rejection, infection time points) dynamic monitoring of immune cell function, at the same time in the same during rejection, infection of transplant recipients, a total of 131 cases, according to the clinical situation of groups: healthy control group (HC), before renal transplantation group (Pre-Tx), postoperative stable group (Tx), postoperative infection group (Infection), rejection group (AR), and acute kidney injury group, postoperative stability, rejection and acute kidney injury group were confirmed by renal biopsy, infection was confirmed by chest X-ray, chest CT and etiological examination confirmed the diagnosis. The anti coagulation heparin from peripheral (2M1), analysis of cellular immune function (ImmuKnow, ATP) and blood 2ml lymphocyte subsets for points Analysis (CD4/CD8), 1ml Fk506 for the detection of blood concentration, 5ml for mixed lymphocyte culture (MLR). The experimental data were analyzed using SPSS 16 software, between the two groups were compared with T test, the rejection group and the other groups compared with One-Way ANONA, Hochberg's GT2, P0.05 was statistically significant, correlation analysis using Logistic regression the analysis method.
The healthy people ATP concentration was 401 + 153ng/ml, preoperative renal transplant recipients ATP concentration was 405 + 161ng/ml, two compared (median), before renal transplantation group (317 ng/ml) ATP concentration was lower than that of group HC (404ng/ml).ImmuKnow showed ATP concentration after renal transplantation immune status fluctuation, after 1 a month when the concentration of ATP (603 + 76ng/ml) 3 months after operation was higher than that of (424 + 191 ng/ml) and 6 months (396 + 175 ng/m1) follow-up value with time stable.AR group concentration of ATP (539 + 217 ng/ml) and other groups (group Tx, 332 + 155 ng/ml, 288 in Infection group. 140 ng/ml, AKI = 336 + 125 ng/ml) were higher than the other groups, the difference was statistically significant (P0.05). The ATP concentration of the recipients with MLR, CD4/CD8 and Fk506 blood concentration respectively to do correlation analysis, found no significant correlation between ATP concentration and the correlation coefficients were three, R2= 0.0072, R2=2E-06. R2=0.004.
Conclusion this research used 1 new immune state monitoring method of cellular immune function (ImmuKnow ATP concentration detection method) cell immune status changes observed after kidney transplantation, the recipient immune state in a strong state within one month after the operation, with the prolongation of time tends to detect stable.2 ImmuKnow method ATP value there is an upward trend in the incidence of acute rejection, acute rejection of the.3ImmuKnow ATP indicates the value of cellular immunity and the direct reaction of CD4/CD8, commonly used in clinical detection methods of MLR, Fk506 blood concentrations were not correlated.

【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R699.2;R392

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