肾移植受者术前供者反应性干扰素γ和转化生长因子β生成细胞的频数及其意义

Pre-transplant donor reactive IFN-γ producing cell and TGF-β producing cell frequencies and their clinical significance

目的检测肾移植受者术前血中淋巴细胞供者反应性干扰素γ（IFN-γ）和转化生长因子β（TGF-β）生成细胞的频数，并探讨与急性排斥反应（AR）和术后6个月移植肾功能的关系。方法分离156例。肾移植受者术前外周血淋巴细胞和尸肾供者脾脏细胞（活体移植为供者外周血淋巴细胞），应用酶联免疫斑点技术（ELISPOT）检测受者术前血中淋巴细胞供者反应性IFN-γ和TGF-γ生成细胞的频数，比较术后AR和非AR患者的差异。采用SPSS11．0软件行t检验和Х^2检验。结果AR患者供者反应性IFN-γ ELISPOT显著高于非AR患者，分别为27．3和19．2（P=0．013）。IFN-γ ELISPOT阳性48例（31％），出现AR28例（58％），阴性108例，出现AR25例（23％，P=0．007）。移植术后6个月IFN-γ ELISPOT阳性组GFR为（53±15）ml／min，阴性组为（73±16）ml／min；2组比较差异有统计学意义（P=0．005）。结论移植前IFN-γ ELISPOT检测可作为肾移植受者移植前的细胞“交叉配型试验”，为进一步避免预致敏的供受者组合提供新手段。术前IFN-γ ELISPOT与肾移植术后6个月肾功能相关，可能为肾移槽噩者远期预后提供信息。

Objective To evaluate the frequencies of pre-transplant donor reactive IFN-γ and TGF-β producing cells using enzyme-linked immunosorbent （ELISPOT） assay and determine their correlation with the diagnosis of acute rejection and kidney graft function at 6 months post transplant. Methods One hundred and fifty-six kidney transplant recipients who were transplanted and followed up were enrolled. ELISPOT measurements of frequencies of donor reactive IFN-γ producing cells and TGF-β producing cells were carried out in pre-transplant peripheral blood lymphocyte （PBL） from the recipients as the responders, and splenocytes from the cadaveric donor or PBLs from live donor as stimulator cells. Patients with or without acute rejection episodes during the follow up were compared for the differences in IFN-γ and TGF-β ELISPOT values. Results Kidney transplant recipients who experienced acute rejection episodes had significantly higher IFN-γ ELISPOT than non-rejectors （mean rank 27.3 vs. 19.2, P=0. 013）. Forty-eight patients were defined as IFN-γ ELISPOT positive （〉 30/300 000）, among whom 28（58%） had developed acute rejection, while only 25/108（23%） negative patients had acute rejection （chi-square, P =0. 007）. Calculated glomerular filtration rate （Cockcroft-Gault） was （73 ± 16） ml/min in IFN-γ ELISPOT negative patients versus （53 ± 15） ml/min in positive patients （P=0. 005）. Conclusions Pre-transplant IFN-γ ELISPOT can be served as a final cellular cross match test for kidney transplant recipients, which may be used by the clinicians to fur-ther improve donor and recipient pairing. Pre-transplant IFN-γ ELISPOT correlated with kidney allograft function at 6 month post transplant, thereby may be a potential predictor for long-term outcomes and guide individualized treatment on a more rational basis.