可溶性CD30和Th1/Th2检测在慢性移植肾功能不全患者免疫状态评估中的应用

Determination of sCD30 and Th1/Th2 in the evaluation of immune status of patients with chronic renal allograft dysfunction

目的探讨将血清可溶性CD30(sCD30)和外周血CD3+CD8-T淋巴细胞Th1/Th2检测用于评估慢性移植肾功能不全(CRAD)患者免疫状态的价值。方法通过分析临床资料,结合移植肾穿刺活检,选取移植术后远期(>6个月)免疫损伤为主的慢性移植肾功能不全患者(A组)、非免疫损伤为主的慢性移植肾功能不全患者(B组)和移植肾功能正常患者(C组)各15例,采用流式细胞术检测每例患者外周血CD3+CD8-T淋巴细胞IFN-γ和IL-4表达率,以IFN-γ和IL-4表达率之比作为Th1/Th2比例,同时留取标本以ELISA法行血清sCD30水平测定。结果A、B、C组患者外周血CD3+CD8-T细胞IFN-γ表达率(11.2%±6.2%、10.9%±6.4%、12.3%±6.9%)无统计学差异(P>0.05)。A组患者外周血CD3+CD8-T细胞IL-4表达率(4.0%±2.8%)明显低于B、C组(7.9%±5.5%、10.2%±7.5%,P<0.01),且其Th1/Th2比例(3.1±1.1)明显高于B、C组(1.5±0.5、1.4±0.5,P<0.01)。B、C组患者外周血CD3+CD8-T细胞IL-4表达率和Th1/Th2比例无统计学差异(P>0.05)。A组患者血清sCD30水平(20.2±12.4ng/ml)明显高于B、C组(7.8±3.1ng/ml、7.6±3.0ng/ml,P<0.01),而B、C组患者血清sCD30水平无统计学差异(P>0.05)。受试者工作特征(ROC)曲线分析显示,当Th1/Th2比例取1.95时,鉴别以免疫损伤为主的CRAD的敏感度为80%,特异度为90%;血清sCD30水平取10.0ng/ml时,鉴别以免疫损伤为主的CRAD的敏感度为93.3%,特异度为86.7%。结论以免疫损伤为主的CRAD患者大都存在Th1/Th2平衡失调(向Th1方向偏移),且血清sCD30水平相对较高。根据外周血CD3+CD8-T细胞Th1/Th2比例和血清sCD30水平鉴别以免疫损伤为主的CRAD具有较高的敏感性和特异性。

Objection To discuss the significance of determining serum sCD30 and Th1/Th2 of CD3＋CD8-T lymphocytes in peripheral blood for evaluating immune status of patients with chronic renal allograft dysfunction（CRAD）.Methods By analyzing the clinical data combined with the result of needle biopsy of the allograft,15 patients with CRAD due mainly to immune injury（group A）,15 patients with CRAD due to non-immune injury（group B）and 15 patients with normal allograft function（group C）in late post-transplantation period（〉6 month）were enrolled for the present study.The expression percentage of IFN-γ and IL-4 of CD3＋CD8-T lymphocytes in peripheral blood of all the 3 groups of patients were determined with flow cytometry（FCM）.The ratio of IFN-γ/IL-4 was taken as the ratio of Th1/Th2.Meanwhile,the serum samples of all patients were collected,and the level of serum sCD30 was determined later by ELISA.Results No significant difference was found in the expression percentage of IFN-γ of CD3＋CD8-T lymphocytes cells in peripheral blood of the patients among all the three groups（11.2%±6.2%,10.9%±6.4% and 12.3%±6.9% for group A,B and C,P〉0.05）.The expression percentage of IL-4 of CD3＋CD8-T lymphocytes cells in peripheral blood of the patients in group A（4.0%±2.8%）was significantly lower than that in group B and C（7.9%±5.5% and 10.2%±7.5%,P〈0.01）.The ratio of Th1/Th2 in the patients of group A（3.1±1.1）was significantly higher than that in the patients of group B and C（1.5±0.5 and 1.4±0.5,P〈0.01）.However,no significant difference was found between group B and C on the expression percentage of IL-4 and the ratio of Th1/Th2 of CD3＋CD8-T lymphocytes cells in peripheral blood（P〉0.05）.The level of serum sCD30 in the patients of group A（20.2±12.4ng/ml）was significantly higher than that of group B and C（7.8±3.1ng/ml and 7.6±3.0ng/ml,P〈0.01）,but no significant difference was found on the levels of serum sCD30 in the patients between group B and C（P〉0.05）.ROC curve analysis indicated that when the ratio of Th1/Th2 was at the cut-off value of 1.95,the sensitivity and specificity to identify CRAD caused mainly by immune injury was 80% and 90%,respectively;and when the level of serum sCD30 was at the cut-off value of 10ng/ml,the sensitivity and specificity to identify CRAD caused mainly by immune injury was 93.3% and 86.7%,respectively.Conclusions Disequilibrium of Th1/Th2（drift to Th1）and raised level of serum sCD30 exist in most of the patients with CRAD which was caused mainly by immune injury.It is with high sensitivity and specificity to identify the CRAD by determining the ratio of Th1/Th2 of CD3＋CD8-T lymphocytes cells in peripheral blood and the level of serum sCD30.