Evaluation of a newly identified soluble CD14 subtype as a marker for sepsis

CD14, a high-affinity receptor for lipopolysaccharide (LPS), is a glycoprotein expressed on the surface membranes of monocytes/macrophages. We have identified a previously unknown form of soluble CD14, named soluble CD14 subtype (sCD14-ST), that is increased in patients with sepsis. To measure sCD14-ST concentrations in plasma, we prepared anti-sCD14-ST antibodies and developed an enzyme immunoassay (EIA) for this soluble form of CD14. With this assay, quantitative measurements are available within 4 h, and we compared the levels of sCD14-ST in plasma from normal subjects (healthy controls), patients with systemic inflammatory response syndrome (SIRS), and sepsis patients. The level of sCD14-ST in subjects with sepsis was much higher than the levels in subjects with SIRS and the healthy controls. Additionally, when a subject's sCD14-ST level was used as a diagnostic marker for sepsis, the area under the receiver operating characteristic (ROC) curve was 0.817, thereby demonstrating that elevated sCD14-ST levels were a better marker for sepsis than the other molecular markers we tested. sCD14-ST levels also correlated with procalcitonin (PCT) levels and with sequential organ failure assessment (SOFA) scores. Finally, changes in sCD14-ST concentration correlated with the severity of sepsis. Taken together, these results indicate that sCD14-ST is a useful marker for the rapid diagnosis of sepsis and for monitoring the severity of the disease.     

可溶性白细胞分化抗原14在脓毒症中的研究进展

脓毒症、严重脓毒症和脓毒症休克是急诊科经常处理的疾病,有很高的病死率,早期诊断是提高生存率的关键因素之一。近几年,降钙素原被用作诊断脓毒症患者的生物学标记物,尽管它与感染密切相关,但是特异性有限,在发生全身炎症反应综合征时会升高。可溶性白细胞分化抗原14(sCD14-st,又称presepsin)是一种存在于单核细胞/巨噬细胞中的糖蛋白。初步报道显示脓毒症患者体内的sCD14-st水平比健康者体内的sCD14-st显著升高。     

重症脓毒症患者血清脂多糖结合蛋白及其受体变化的临床研究

目的观察重症脓毒症患者血清脂多糖结合蛋白（LBP）及其受体CD14（CD14）的变化，探讨两者与重症脓毒症严重程度及预后的关系。方法采用酶联免疫吸附法（ELISA）测定41例重症脓毒症患者发病后不同时间血清LBP和可溶性CD14（sCD14）浓度以及发病后24h内血清前降钙素（PCT）和内毒索（LPS）浓度，并且进行相关性分析。结果重症脓毒症患者发病后各时间点血清LBP和sCD14水平较正常对照均显著升高（P均〈0．01）；死亡组患者各时间点血清LBP水平与存活组间差异均无显著性（P均〉0．05），而sCD14则均较存活组显著升高（P均〈0．05）。血清LBP水平与PCT、急性生理学与慢性健康状况评分Ⅱ（APACHEⅡ）无显著相关性，与LPS呈显著正相关；而sCD14与PCT、APACHEⅡ评分呈显著正相关，与LPS无显著相关性。结论重症脓毒症患者血清LBP和sCD14浓度均显著升高，LBP仅反映机体急性炎症反应，而不能作为判断疾病严重程度及预后的指标；而sCD14升高程度可提示预后，在一定程度上反映脓毒症的严重程度。     

脑梗塞患者血清可溶性CD14水平的初步观察

目的　通过检测脑梗塞患者血清可溶性CD14 (sCD14 )水平,研究缺血性脑血管病患者外周血单核细胞的活化状态。方法　收集脑梗塞患者和正常人血清,用单多抗双夹心ELISA法检测血清sCD14含量并进行统计分析。结果　脑梗塞患者血清sCD14水平较同年龄正常人显著升高(P <0. 0 0 1) ,其中感染组、未感染组血清sCD14含量较正常人均显著升高(P值分别为0 .0 0 1和0 . 0 11) ;而感染组与未感染组之间血清sCD14含量无显著性差异(P =0 .175 ) ;血清sCD14含量与白细胞数无显著相关性(P =0. 17)。结论　脑梗塞患者血清sCD14水平升高,提示其外周血单核细胞处于活化状态,但与是否存在感染无明显相关性。     

冠心病患者sCD14血清水平检测的临床意义

目的探讨血清可溶性CD14(sCD14)水平在监测冠心病病情及其与冠状动脉病变程度的关系。方法168例冠心病患者,按临床诊断分为三组:急性心肌梗死(AMI)60例、不稳定型心绞痛(UAP)58例、稳定型心绞痛(SAP)50例和对照组55例。用酶联免疫吸附试验检测各组血清sCD14的水平,并比较各组间的差异。对冠心病患者的外周血白细胞总数变化及其与血清sCD14水平进行直线相关分析。结果AMI组、UAP组及SAP组的血清sCD14水平比对照组高(P＜0．05);AMI组、UAP组sCD14水平和SAP组相比,其值增加明显;AMI组和UAP组结果相似;冠心病患者sCD14水平与外周血白细胞总数变化呈正相关。结论sCD14可能是冠状动脉粥样硬化的标志,参与了冠心病的发病过程,其值与冠状动脉病变程度密切相关。     

Presepsin (soluble CD14 subtype) and procalcitonin levels for mortality prediction in sepsis: data from the Albumin Italian Outcome Sepsis trial

Introduction

Sepsis, a leading cause of death in critically ill patients, is the result of complex interactions between the infecting microorganisms and the host responses that influence clinical outcomes. We evaluated the prognostic value of presepsin (sCD14-ST), a novel biomarker of bacterial infection, and compared it with procalcitonin (PCT).

Methods

This is a retrospective, case–control study of a multicenter, randomized clinical trial enrolling patients with severe sepsis or septic shock in ICUs in Italy. We selected 50 survivors and 50 non-survivors at ICU discharge, matched for age, sex and time from sepsis diagnosis to enrollment. Plasma samples were collected 1, 2 and 7 days after enrollment to assay presepsin and PCT. Outcome was assessed 28 and 90 days after enrollment.

Results

Early presepsin (day 1) was higher in decedents (2,269 pg/ml, median (Q1 to Q3), 1,171 to 4,300 pg/ml) than in survivors (1,184 pg/ml (median, 875 to 2,113); P = 0.002), whereas PCT was not different (18.5 μg/L (median 3.4 to 45.2) and 10.8 μg/L (2.7 to 41.9); P = 0.31). The evolution of presepsin levels over time was significantly different in survivors compared to decedents (P for time-survival interaction = 0.03), whereas PCT decreased similarly in the two groups (P = 0.13). Presepsin was the only variable independently associated with ICU and 28-day mortality in Cox models adjusted for clinical characteristics. It showed better prognostic accuracy than PCT in the range of Sequential Organ Failure Assessment score (area under the curve (AUC) from 0.64 to 0.75 vs. AUC 0.53 to 0.65).

Conclusions

In this multicenter clinical trial, we provide the first evidence that presepsin measurements may have useful prognostic information for patients with severe sepsis or septic shock. These preliminary findings suggest that presepsin may be of clinical importance for early risk stratification.     

Association between serum soluble CD40 ligand levels and mortality in patients with severe sepsis

Introduction

CD40 Ligand (CD40L) and its soluble counterpart (sCD40L) are proteins that exhibit prothrombotic and proinflammatory properties on binding to their cell surface receptor CD40. The results of small clinical studies suggest that sCD40L levels could play a role in sepsis; however, there are no data on the association between sCD40L levels and mortality of septic patients. Thus, the aim of this study was to determine whether circulating sCD40L levels could be a marker of adverse outcome in a large cohort of patients with severe sepsis.

Methods

This was a multicenter, observational and prospective study carried out in six Spanish intensive care units. Serum levels of sCD40L, tumour necrosis factor-alpha and interleukin-10, and plasma levels of tissue factor were measured in 186 patients with severe sepsis at the time of diagnosis. Serum sCD40L was also measured in 50 age- and sex-matched controls. Survival at 30 days was used as the endpoint.

Results

Circulating sCD40L levels were significantly higher in septic patients than in controls (P = 0.01), and in non-survivors (n = 62) compared to survivors (n = 124) (P = 0.04). However, the levels of CD40L were not different regarding sepsis severity. Logistic regression analysis showed that sCD40L levels >3.5 ng/mL were associated with higher mortality at 30 days (odds ratio = 2.89; 95% confidence interval = 1.37 to 6.07; P = 0.005). The area under the curve of sCD40L levels >3.5 ng/mL as predictor of mortality at 30 days was 0.58 (95% CI = 0.51 to 0.65; P = 0.03).

Conclusions

In conclusion, circulating sCD40L levels are increased in septic patients and are independently associated with mortality in these patients; thus, its modulation could represent an attractive therapeutic target.     

Diagnostic accuracy of presepsin (soluble CD14 subtype) for prediction of bacteremia in patients with systemic inflammatory response syndrome in the Emergency Department

BackgroundBacteremia is indicative of severe bacterial infection with significant mortality. Its early diagnosis is extremely important for implementation of antimicrobial therapy but a diagnostic challenge. Although blood culture is the “gold standard” for diagnosis of bacteremia this method has limited usefulness for the early detection of blood-stream infection. In this study we assessed the presepsin as predictor of bacteremia in patients with systemic inflammatory response syndrome (SIRS) on admission to the Emergency Department and compare it with current available infection biomarkers.MethodsA total of 226 patients admitted to the Emergency Department with SIRS were included. In 37 patients blood culture had a positive result (bacteremic SIRS group) and 189 had a negative blood culture result (non-bacteremic SIRS group). Simultaneously with blood culture, presepsin, procalcitonin (PCT) and C-reactive protein (CRP) were measured. Receiver operating characteristic (ROC) curve analysis was performed for each biomarker as predictor of bacteremia.ResultsPresepsin values were significantly higher in bacteremic SIRS group when compared with non-bacteremic SIRS group. ROC curve analysis and area under curve (AUC) revealed a value of 0.750 for presepsin in differentiating SIRS patients with bacteremia from those without, similar than that for PCT (0.787) and higher than that for CRP (0.602). The best cut-off value for presepsin was 729 pg/mL, which was associated with a negative predictive value of 94.4%.ConclusionPresepsin may contribute to rule out the diagnosis of bacteremia in SIRS patients admitted to the Emergency Department.     

Diagnostic and prognostic utility of soluble CD 14 subtype (presepsin) for severe sepsis and septic shock during the first week of intensive care treatment

Introduction

The aim of this study was to evaluate the diagnostic and prognostic value of presepsin in patients with severe sepsis and septic shock during the first week of ICU treatment.

Methods

In total, 116 patients with suspected severe sepsis or septic shock were included during the first 24 hours of ICU treatment. Blood samples for biomarker measurements of presepsin, procalcitonin (PCT), interleukin 6 (IL-6), C reactive protein (CRP) and white blood cells (WBC) were drawn at days 1, 3 and 8. All patients were followed up for six months. Biomarkers were tested for diagnosis of sepsis, severe sepsis, septic shock and for prognosis of 30-days and 6-months all-cause mortality at days 1, 3 and 8. Diagnostic and prognostic utilities were tested by determining diagnostic cutoff levels, goodness criteria, C-statistics and multivariable Cox regression models.

Results

Presepsin increased significantly from the lowest to most severe sepsis groups at days 1, 3 and 8 (test for linear trend P <0.03). Presepsin levels revealed valuable diagnostic capacity to diagnose severe sepsis and septic shock at days 1, 3 and 8 (range of diagnostic area under the curves (AUC) 0.72 to 0.84, P = 0.0001) compared to IL-6, PCT, CRP and WBC. Goodness criteria for diagnosis of sepsis severity were analyzed (≥sepsis, cutoff = 530 pg/ml; ≥severe sepsis, cutoff = 600 pg/ml; ≥septic shock, cutoff = 700 pg/ml; P <0.03). Presepsin levels revealed significant prognostic value for 30 days and 6 months all-cause mortality (presepsin: range of AUC 0.64 to 0.71, P <0.02). Patients with presepsin levels of the 4^th quartile were 5 to 7 times more likely to die after six months than patients with lower levels. The prognostic value for all-cause mortality of presepsin was comparable to that of IL-6 and better than that of PCT, CRP or WBC.

Conclusions

In patients with suspected severe sepsis and septic shock, precipices reveals valuable diagnostic capacity to differentiate sepsis severity compared to PCT, IL-6, CRP, WBC. Additionally, presepsin and IL-6 reveal prognostic value with respect to 30 days and 6 months all-cause mortality throughout the first week of ICU treatment.

Trial registration

ClinicalTrials.gov {"type":"clinical-trial","attrs":{"text":"NCT01535534","term_id":"NCT01535534"}}NCT01535534. Registered 14 February 2012.     

严重烧伤并发肝损害时可溶性脂多糖受体CD14的表达

严重烧伤患者常并发肝功能损害 ,这与烧伤后的内毒素血症 (ETM)关系密切。体外实验发现 ,脂多糖结合蛋白(L BP) /脂多糖受体 (CD14 )在脂多糖(L PS)的识别和细胞应答反应中具有重要的调节作用 ,可能是增敏内毒素细胞效应的重要系统之一 〔1〕,但严重烧伤患者可溶性脂多糖受体 CD14 (s CD14 )的变化及其与 L PS诱发肝损害的关系不十分清楚。为此我们进行了研究 ,探讨s CD14在严重烧伤后肝损害中的意义。1　资料与方法1.1　病例 :烧伤面积大于 5 0 %患者 2 0例 ,男 13例 ,女 7例 ;平均年龄 (2 9.2±6 .5 )岁 ;皆为热力烧伤 ,无中度以…     

Evaluation of sepsis in a critically ill surgical population

We report a new clinical rating system which assesses septic patients' ongoing disease course and its severity. Our system incorporates the Therapeutic Intervention Scoring System (TISS) and Acute Physiology and Chronic Health Evaluation to measure discrete organ system abnormalities, plus a multiple system organ failure scale to quantify the number of abnormal organ systems. The resulting score, which reflects the severity of multiple organ dysfunction and grades responsiveness to therapy, was validated against the actual disease course. Retrospective and prospective profiles of individual surgical ICU patients demonstrated that this tracking method was a more effective indicator of severity of sepsis and more sensitive to the day-to-day changes in clinical status than either the TISS or APACHE II components alone. We also demonstrate that a graphic illustration of daily system scores yields clinically useful information relevant to the patients' septic course.     

Lipopolysaccharide-binding protein,lipopolysaccharide, and soluble CD14 in sepsis of critically ill neonates and children

OBJECTIVE: To compare the diagnostic accuracy of lipopolysaccharide-binding protein (LBP) for sepsis in critically ill neonates and children with the two markers participating in the same inflammatory pathway, lipopolysaccharide and soluble CD14. DESIGN AND SETTING: Prospective, observational study in a multidisciplinary neonatal and pediatric intensive care unit. PATIENTS: 47 critically ill neonates and 49 critically ill children with systemic inflammatory response syndrome (SIRS) and suspected sepsis, classified into two groups: those with and those without sepsis. INTERVENTIONS: Serum LBP, lipopolysaccharide, soluble CD14, C-reactive protein, and procalcitonin were measured on 2 consecutive days. The area under the receiver operating characteristic curve (AUC), sensitivity, specificity, and predictive values were evaluated. RESULTS: AUC for LBP on the first day of suspected infection was 0.97 in neonates aged under 48 h, 0.93 in neonates over 48 h and 0.82 in children. AUCs for lipopolysaccharide and soluble CD14 were 0.77 and 0.74 in neonates under 48 h, 0.53 and 0.76 in neonates over 48 h, and 0.72 and 0.53 in children. AUCs for procalcitonin and C-reactive protein were 0.65 and 0.89 in neonates under 48 h, 0.65 and 0.91 in neonates over 48 h, and 0.76 and 0.69 in children. CONCLUSIONS: In critically ill neonates and children LBP concentration on the first day of suspected sepsis is a better marker of sepsis than lipopolysaccharide, soluble CD14, procalcitonin, and in neonates younger than 48 h and children, also a better marker than C-reactive protein. Lipopolysaccharide and soluble CD14 are not suitable markers for the differentiation of infectious and noninfectious SIRS.     

Platelet and soluble CD40L in meningococcal sepsis

Objective To determine the influence of meningococcal sepsis on levels of platelet derived CD40L and on endothelial CD40 expression.Design and setting Prospective observational study in two tertiary paediatric intensive care units.Patients and participants 63 children with meningococcal sepsis and 10 age-matched controls.Measurements and results (a) sCD40L ELISA of plasma from patients with meningococcal sepsis (n = 49) and age matched controls (n = 10). This demonstrated higher sCD40L levels in patients (median 0.29 ng/ml, IQR 0.2–0.41) than controls (0.09 ng/ml, 0.08–0.12). However, there was no relationship between plasma sCD40L level and platelet count or disease severity. (b) Flow cytometry of fresh blood from patients with meningococcal sepsis (n = 11) and age-matched controls (n = 10) for membrane bound CD40L and CD62P on circulating platelets. This demonstrated low levels of CD40L and CD62P in patients and controls. CD40L⁺ platelets were 3.5% (3.0–4.8) in patients and 4.9% (3.5–4.3) in controls. CD62P⁺ platelets were 10.7% (6.4–12.8) in patients and 7.9% (5.9–13.0) in controls. (c) Immunohistochemistry of skin biopsy specimens from six patients, staining for endothelial CD40 expression at sites of microthrombus formation, which demonstrated preserved CD40 expression in vascular endothelium at sites of microthrombus formation.Conclusions The elevated sCD40L level in meningococcal sepsis implies release of sCD40L from platelets. However, there was no relationship between plasma sCD40L level and the degree of thrombocytopenia or disease severity. Furthermore, platelet surface bound CD40L was similar in controls and patients. Thus, further investigation is needed to determine whether platelet CD40L contributes to inflammation and thrombosis in MCS.D.P.I. and S.N.F. were supported by the Medical Research Council, UK     

CD14 receptor occupancy in severe sepsis: results of a phase I clinical trial with a recombinant chimeric CD14 monoclonal antibody (IC14)

OBJECTIVE: Binding of bacterial cell wall components to CD14 and co-receptors on myeloid cells results in cellular activation and production of proinflammatory mediators. A recombinant anti-CD14 monoclonal antibody (IC14) has been shown to decrease lipopolysaccharide-induced responses in animal and human models of endotoxemia. This study was performed to evaluate the safety, pharmacokinetics, pharmacodynamics, and clinical pharmacology of IC14 in patients with severe sepsis. DESIGN: Randomized, double-blind, placebo-controlled, dose-ranging, multiple-center trial. SETTING: Six medical and surgical intensive care units located in Germany and The Netherlands. PATIENTS: Forty patients with severe sepsis. INTERVENTIONS: IC14 was administered intravenously to eight patients/cohort as single (1 mg/kg or 4 mg/kg) or multiple doses (4 mg/kg daily for 4 days, or 4 mg/kg on day 1 followed by 2 mg/kg daily for 3 days). A placebo group (two patients/cohort) was also included. MEASUREMENTS AND MAIN RESULTS: The overall incidence and types of adverse events were similar among treatment groups. One patient in the group receiving multiple-dose IC14 4 mg/kg daily for 4 days experienced an anaphylactic reaction after receiving the first dose of study drug. IC14 did not induce antibody formation or increase the incidence of secondary bacterial infection. A mean IC14 serum concentration of approximately 1 microg/mL was required to achieve 50% of maximum membrane-bound CD14 receptor occupancy on peripheral blood monocytes. The pattern of proinflammatory and anti-inflammatory cytokines, chemokine, soluble receptor, soluble E-selectin, and acute phase proteins in response to treatment was highly variable by patient and IC14 treatment group. CONCLUSIONS: Single and multiple doses of IC14 were generally well tolerated and did not induce antibody formation or increase the incidence of secondary bacterial infection. The results suggest that CD14 blockade with IC14 warrants further clinical investigation to determine its ability to attenuate the proinflammatory response due to infection.     

脑卒中患者血清可溶性CD14、同型半胱氨酸和7项血脂结果分析

目的 探讨血清中可溶性CD14(sCD14)、同型半胱氨酸(HCY)和7项血脂指标与脑卒中的相关性. 方法 采用ELISA法检测195例脑卒中患者及120例健康对照组血清sCD14;使用罗氏C7O2全自动生化分析仪检测195例脑卒中患者及120例健康对照组血清HCY和7项血脂,并对检测结果进行分析. 结果 脑卒中患者中sCD14、HCY、TG、CHOL、HDL、LDL-C、Apo A、Apo B、LP(a)含量分别为(79.98±36.67)μg/mL、(20.78± 17.98) μmol/L、(1.48±0.78)mmol/L、(5.19±0.93)mmol/L、(1.15±0.31)mmol/L、(2.62±0.83)mmol/L、(1.34±0.26) g/L、(0.97±0.76) g/L、(188.07±167.68)mg/L;脑卒中患者血清sCD14、HCY、TG、CHOL、LDL-C含量明显升高,与健康对照组比较,差异具有统计学意义(P＜0.05),而脑卒中患者HDL、Apo A、Apo B、LP(a)含量与健康对照组比较,差异无统计学意义(P＞0.05).结论 脑卒中患者血清sCD14、HCY和TG、CHOL、LDL-C明显相关,对了解患者的发病和辅助诊断具有重要的临床意义.     

人可溶性CD14检测体系的建立

目的　建立人可溶性CD14双抗体夹心ELISA检测体系。方法　用抗人CD14单抗为捕获抗体 ,加入待测抗原 ,第二抗体为兔抗人CD14多克隆抗体 ,最后加入辣根过氧化物酶标记的羊抗兔IgG。结果　本检测体系灵敏度高 ,可达 5ng ml,特异性强 ,测量范围 5～ 12 0ng ml,重复性好 ,各项技术指标与国外同类产品相当。结论　本检测体系能够检测人体液中和细胞培养上清中sCD14 ,适用于临床及基础研究。     

乌司他丁对重症脓毒症患者脂多糖结合蛋白及其受体的影响

目的观察重症脓毒症患者血清LBP及sCD14的变化规律,探讨尿胰蛋白酶抑制剂的作用及可能的作用机制。方法用酶联免疫吸附（EusA）法测定40例重症脓毒症患者各时点的血清LBP和sCD14浓度。将其随机分为乌司他丁组（U组）与对照组（C组）,在相同的常规治疗基础上,U组给予乌司他丁针剂20万u静脉注射,2次／d,持续5d。C组则给予同等量的生理盐水作为安慰剂对照。在治疗前、治疗后第2,3,6天抽取右侧桡动脉血测定血清LBP及sCD14的浓度,观察其动态变化的规律,并统计两组患者28d的病死率。结果重症脓毒症患者在诊断成立的第2天血清LBP及sCD14水平达高峰并随后回落,第6天LBP水平仍高于正常而sCD14水平则降至正常范围。死亡组血清sCD14水平较存活组有升高,在第6天差异才具有统计学意义。治疗前两组患者APACHEI／评分值相似（P〉0．05）,U组患者28d的病死率为18．2％（4／22）。C组为50．0％（9／18）,差异具有统计学意义（P〈0．05）。治疗5d后,血清sCD14水平在U组有下降（P〈0．05）。结论血清sCD14水平可以作为判断重症脓毒症患者预后的指标之一;尿胰蛋白酶抑制剂能够改善脓毒症患者的预后,降低病死率,可能与其改变脓毒症患者血清中sCD14的水平有关。