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目的探讨检测CD4+T淋巴细胞内三磷腺苷(ATP)水平用于评价肾移植受者细胞免疫功能的价值。方法采用植物血凝素刺激、单克隆抗体包被免疫磁珠分离CD4+T淋巴细胞,以荧光试剂标记,测定27例肾移植受者(研究组)的术前,术后3d、7d、14d、30d以及发生感染时的CD4+T淋巴细胞内ATP水平,并以14名健康人作对照(对照组)。研究组患者术后均采用抗人T细胞CD3鼠单抗(OKT3)诱导治疗,并予免疫抑制剂治疗,其中21例采用以环孢素(CsA)为基础的三联用药方案,6例采用以他克莫司(FK506)为基础的三联用药方案,比较采用不同用药方案患者的ATP水平,采用Pearson相关分析评估肾移植患者ATP水平与环孢素或他克莫司血药浓度的关系。结果肾移植受者术前CD4+T淋巴细胞内ATP水平为(382±187)ng/ml,明显低于正常成人的(496±116)ng/ml(P<0.05),术后7d降至最低,14d开始回升,30d时仍低于术前水平(P<0.01)。3例术后并发肺部感染者,其CD4+T淋巴细胞内ATP水平下降,病情加重时下降更加明显,感染控制后有所回升。采用环孢素三联方案治疗的患者各时点的ATP水平与采用他克莫司三联方案治疗的患者比较差异无统计学意义。肾移植受者的ATP水平与上述两药血药浓度变化无关。结论测定肾移植受者CD4+T淋巴细胞内ATP水平可反映患者的整体细胞免疫状态,指导临床制定个体化治疗方案。
Objective To investigate the value of detecting adenosine triphosphate (ATP) in CD4 + T lymphocytes to evaluate cellular immune function in renal allograft recipients. Methods Phytohemagglutinin was used to stimulate CD4 (superscript +) T lymphocytes by immunomagnetic beads coated with monoclonal antibody and labeled with fluorescent reagent. The preoperative, postoperative 3d, 7d, 14d postoperatively in 27 renal transplant recipients (study group) , 30d and the level of ATP in CD4 + T lymphocytes at the time of infection, and 14 healthy people as control (control group). All patients in the study group were treated with anti-human T cell CD3 murine monoclonal antibody (OKT3) and immunosuppressed. 21 cases were treated with cyclosporine (CsA) -based triple therapy and 6 cases were treated with Tacrolimus (FK506) -based triple-drug regimen was used to compare the ATP levels of patients treated with different regimens. Pearson correlation analysis was used to assess the relationship between ATP levels and plasma concentrations of cyclosporine or tacrolimus in renal transplant recipients. Results The level of ATP in CD4 + T lymphocytes was (382 ± 187) ng / ml preoperatively in renal transplant recipients, which was significantly lower than that in normal adults (496 ± 116 ng / ml, P <0.05) The lowest, 14d began to rise, still below the preoperative level 30d (P <0.01). In 3 cases with postoperative pulmonary infection, the level of ATP in CD4 + T lymphocytes decreased, and the decline was more obvious when the condition was aggravated. There was no significant difference in the level of ATP at different time points between patients treated with cyclosporin triple therapy and those treated with tacrolimus triple therapy. The level of ATP in renal transplant recipients has nothing to do with the change of plasma concentrations of these two drugs. Conclusion The determination of ATP level in CD4 + T lymphocytes in renal transplant recipients can reflect the overall cellular immune status of patients and guide the clinical development of individualized treatment regimens.