外周血单核细胞亚群及CD64表达水平对脓毒症诊断和预后评估的价值

Value of peripheral blood monocyte subsets and CD64 expression in the diagnosis and prognosis of sepsis

目的探讨单核细胞亚群及CD64表达水平在脓毒症诊断和预后评估中的价值。方法采用前瞻性病例对照研究设计,选择2021年3月至2022年3月由解放军陆军特色医学中心重症监护病房(ICU)连续收治的30例脓毒症和30例非脓毒症患者作为研究对象。采集患者入ICU后1、3、5 d外周血,用流式细胞仪检测单核细胞亚群比例和表面CD64表达水平,分析其在脓毒症与非脓毒症患者中的表达差异;采用单因素及多因素Logistic回归分析筛选脓毒症发生的危险因素;采用受试者工作特征曲线(ROC曲线)评估各危险因素对脓毒症的诊断效能。结果入ICU后1 d,脓毒症患者的单核细胞及经典型单核细胞在白细胞(WBC)中的比例显著低于非脓毒症患者〔单核细胞占WBC比例:(4.13±2.03)%比(6.53±3.90)%,经典型单核细胞占WBC比例:1.97(1.43,2.83)%比3.37(1.71,5.98)%,均P<0.05〕;非经典型单核细胞在单核细胞中的比例显著高于非脓毒症患者〔(11.42±9.19)%比(6.57±4.23)%,P<0.05〕;CD64在单核细胞、经典型单核细胞、中间型单核细胞及非经典型单核细胞上的表达水平均显著高于非脓毒症患者〔平均荧光强度(MFI):单核细胞为13.10±6.01比9.84±2.83,经典型单核细胞为13.58±5.98比10.03±2.84,中间型单核细胞为13.48±6.35比10.22±2.99,非经典型单核细胞为8.21±5.52比5.79±2.67,均P<0.05〕。多因素Logistic回归分析显示,经典型单核细胞CD64〔优势比(OR)=1.299,95%可信区间(95%CI)为1.027~1.471,P=0.025〕和非经典型单核细胞占单核细胞比例(OR=1.348,95%CI为1.034~1.758,P=0.027)是脓毒症发生的独立危险因素。ROC曲线分析显示,经典型单核细胞CD64、非经典型单核细胞占单核细胞比例联合降钙素原(PCT)诊断脓毒症的ROC曲线下面积(AUC)为0.871。相关性分析显示,患者入ICU 1 d经典型单核细胞CD64表达水平与入ICU 1、3、5 d急性生理学与慢性健康状况评分Ⅱ(APACHEⅡ)呈负相关(r值分别为:-0.264、-0.428、-0.368,均P<0.05)。结论外周血经典型单核细胞CD64表达、非经典型单核细胞占单核细胞比例及血浆PCT水平3者联合对脓毒症诊断具有较好的效能,并能评估脓毒症患者的病情严重程度。

Objective To explore the value of monocyte subsets and CD64 expression in the diagnosis and prognosis of sepsis.Methods A prospective case-control study was designed.30 septic patients and 30 non-septic patients who were admitted to the intensive care unit(ICU)of the PLA Army Characteristic Medical Center from March 2021 to March 2022 were enrolled.After 1,3,and 5 days of ICU admission,peripheral blood samples were taken from patients.Flow cytometry was used to detect the proportion of monocyte subsets and the expression level of CD64 on the surface,and the difference of expression between patients in two group was analyzed.The risk variables for sepsis were analyzed using single-factor and multi-factor Logistic regression.The diagnostic efficacy of each risk factor for sepsis was determined using the receiver operator characteristic curve(ROC curve).Results One day after ICU admission,the proportions of monocytes and classic monocytes in white blood cells(WBC)of septic patients were significantly lower than those of non-septic patients[proportion of monocytes to WBC:(4.13±2.03)%vs.(6.53±3.90)%,proportion of classic monocytes to WBC:1.97(1.43,2.83)%vs.3.37(1.71,5.98)%,both P<0.05].The proportion of non-classical monocytes in monocytes was significantly higher in septic patients than that in non-septic patients[(11.42±9.19)%vs.(6.57±4.23)%,P<0.05].The levels of CD64 expression in monocytes,classic monocytes,intermediate monocytes and non-classic monocytes were significantly higher in sepsis patients than those in non-septic patients[mean fluorescence intensity(MFI):13.10±6.01 vs.9.84±2.83 for monocytes,13.58±5.98 vs.10.03±2.84 for classic monocytes,13.48±6.35 vs.10.22±2.99 for intermediate monocytes,8.21±5.52 vs.5.79±2.67 for non-classic monocytes,all P<0.05].Multivariate Logistic regression research showed that CD64 in typical monocytes[odds ratio(OR)=1.299,95%confidence interval(95%CI)was 1.027-1.471,P=0.025]and the proportion of non-typical monocytes in monocytes(OR=1.348,95%CI was 1.034-1.758,P=0.027)were the independent risk factors for sepsis.ROC curve showed that the area under the ROC curve(AUC)of CD64 expression of classical monocytes,the fraction of non-classical monocytes in monocytes,and procalcitonin(PCT)in the diagnosis of sepsis was 0.871.A correlation analysis revealed a negative relationship between the acute physiology and chronic health status evaluationⅡ(APACHEⅡ)on the first,third,and fifth days following ICU admission and the expression level of CD64 in patients'classic monocytes(r values were-0.264,-0.428 and-0.368,respectively,all P<0.05).Conclusions Combining the proportion of non-classical monocytes in monocytes,the level of plasma PCT,and the CD64 expression of classic monocytes in peripheral blood has good efficacy in identifying sepsis and assessing its severity.