Cylex ImmuKnow法检测细胞免疫功能在肾移植后感染诊断中的意义

摘要

目的探讨Cylex ImmuKnow法在临床应用的可行性以及在肾移植术后感染中的诊断价值。方法以48例肾移植术后患者为研究对象，以是否发生感染分为感染组（19例）及非感染组（29例），16例健康成人体检者作为对照组。肾移植术后常规应用激素+钙调蛋白阻滞剂（CNI）〔环孢素A（CsA）或他克莫司（FK506）〕+吗替麦考酚酯（MMF）三联免疫抑制治疗，围手术期应用抗人T细胞免疫球蛋白免疫诱导治疗。采用植物血凝素（PHA）刺激、单克隆抗体包被免疫磁珠分离CD4+T淋巴细胞，荧光试剂标记，测定感染发生前、发生感染时及感染恢复后1周CD4+T淋巴细胞内三磷酸腺苷（ATP）水平。结果肾移植受者发生感染时，CD4+T淋巴细胞内ATP水平明显低于同期非感染患者〔（134.20±58.24）μg/L比（306.32±107.62）μg/L，P＜0.05〕，且较感染发生前明显下降〔（134.20±58.24）μg/L比（359.37±81.35）μg/L， P＜0.01〕，在感染恢复后则上升至感染发生前的水平。结论应用Cylex ImmuKnow检测肾移植受者外周血CD4+T淋巴细胞内ATP水平可反映移植受者的整体细胞免疫状态，为肾移植术后并发感染的诊断和治疗提供客观依据，指导制定个体化免疫抑制方案。%Objective To investigate the feasibility of Cylex ImmuKnow assay in clinical application and its diagnostic value for the infection after renal transplantation. Methods We collected 48 renal transplant recipients. The patients were divided into two groups according to clinical status,namely,infection group(19 cases),and non-infection group(29 cases). Sixteen healthy adults were collected as control. Triple immunosuppressive therapy post renal transplantation consisted of prednisone,calcineurin inhibitors(tacrolimus of cyclosporin)and MMF,with anti-human T lymphocyte rabbit immunoglobulin(ATG)for the immunological induction during perioperative period as well. The adenosine triphosphate(ATP)level within the CD4+ T lymphocytes was detected through labeling the monoclonal antibody coated and immunomagnetic beads isolated CD4+T lymphocytes using fluorescent agents after stimulation by PHA before,onset and one week's recovery after the infection. Results When infection occurred, ATP concentrations in CD4+ T cells of the kidney transplant recipients were significantly lower than those in non-infection group〔(134.20±58.24)μg/L vs.(306.32±107.62)μg/L,P<0.05〕,and those before infection onset〔(134.20±58.24)μg/L vs. (359.37±81.35)μg/L,P<0.01〕. After the infection was fully defeated,the ATP concentrations in CD4+ T cells increased to the level before infection occurred. Conclusion The examination of ATP in CD4+ T cells by Cylex ImmuKnow assay could reflect the status of cellular immunity,provide reliable and objective basis for the diagnosis and treatment of infection after renal transplantation,and guide the clinical individualized immunosuppressive therapy.