Plasma cytokines as predictors of coronary heart disease

BACKGROUND AND OBJECTIVE: Growing body of evidence explicitly suggests the significant role of inflammatory processes in vascular diseases related to atherosclerosis. Monocytes, present in every phase of atherogenesis, are the principal cells accumulating in atherosclerotic plaque. Monocyte Chemotactic Protein 1 (MCP-1) seems to influence firm adherence of rolling monocytes and infiltration into the artery wall. Although the significant meaning of inflammation in atherogenesis has been proved, potential role of antiinflammatory cytokines remains unknown. Interleukin 10 (IL-10) is a major cytokine of pleiotropic antiinflammatory function known to exert inhibitory effects on monocytes. Recent data emerging from clinical and pathological studies suggest important role of thrombosis and fibrinolytic disorders in atherosclerosis complications especially in coronary heart disease (CHD). Individuals with greater Plasminogen Activator Inhibitor 1 (PAI-1) level are believed to be more susceptible to cardiovascular disease. METHODS: In our study we measured the plasma levels of MCP-1, IL-10 and PAI-1 in 10 patients with stable angina and 10 healthy subjects. We also estimated its mutual correlations. The plasma levels of MCP-1, IL-10 and PAI-1 were determined with R&D kits (ELISA). RESULTS: Plasma levels of MCP-1 were significantly higher (261.5+/-40.7 pg/mL vs 73.3+/-3.05 pg/mL; p<0.0002) and also levels of PAI-1 were higher (79.36+/-5.8 ng/mL vs 35.88+/-1.38 ng/mL; p<0.0001) in patients with SA compared with the healthy control subjects. Whereas plasma levels of IL-10 were lower (11.6+/-0.5 pg/mL vs 16.5+/-0.4 pg/mL; p<0.0001) compared with control group and correlated with both MCP-1 plasma level (r=-0.67; p<0.0015) and PAI-1 concentration (r=-0.69; p<0.0008). CONCLUSION: The data obtained confirm the predictive role of cytokines in patients with stable coronary heart disease. The negative correlation of anti-inflammatory IL-10 and PAI-1 was also found.     

Thrombin-antithrombin III complex, proinflammatory cytokines, and fibrinolytic indices for assessing the severity of inflammation in pleural effusions

This study investigated coagulation-related variables, proinflammatory cytokines, and fibrinolytic indices to assess the severity of inflammation in patients with pleural effusions. Tuberculous pleural fluids revealed significantly higher concentrations of tumor necrosis factor-α (TNF-α) and plasminogen activator inhibitor type I (PAI-1) than did malignant and pneumonic pleural fluids. Among the coagulation-related variables, thrombin-antithrombin III complex (TAT) exhibited the largest difference in mean values between pleural fluids and blood samples (125.4 ± 45.1 vs 14.3 ± 20.3 ng/ml, p <0.05). Inflammatory parameters were more closely associated with TAT than tissue type plasminogen activator (tPA), PAI-1, and D-dimers. TAT levels in the severe inflammation group (153.8 ± 45.6 ng/ml) were significantly above those in the mild inflammation group (105.6 ± 38.5 ng/ml, p <0.05); however, no significant differences were observed in PAI-1 and D-dimers levels between the two groups. In conclusion, TNF-α and PAI-1 are important indicators in patients with tuberculous pleural effusions, and measurement of TAT is useful for assessing the severity of inflammation in pleural fluids.     

Urinary 11-dehydro-thromboxane B(2) and coagulation activation markers measured within 24 h of human acute ischemic stroke.

The aim of this study was to determine the extent of change in platelet and coagulation markers in the acute phase of ischemic stroke and to assess the utility of marker measurement in stroke subtype classification. Urinary 11-dehydro-thromboxane B(2) (11-dTXB2), a marker of in vivo platelet activation, and markers of coagulation activation, including prothrombin fragment 1+2 (F1+2), thrombin-antithrombin complex (TAT), and fibrinogen, were measured in 25 patients with ischemic stroke within 24 h of onset of symptoms. Marker levels in patients with ischemic stroke were compared with those in 19 age-matched controls who had not taken aspirin for at least 2 weeks before sampling and 25 healthy controls. Median marker levels were significantly increased in stroke over those in age-matched controls for fibrinogen (344 vs. 289 mg/dl; P=0.030), F1+2 (1.40 vs. 0.80 nmol/l; P=0.003), and TAT (6.65 vs. 2.20 microg/l; P<0.0001). Median marker levels for seven patients with cardioembolic stroke and 18 with non-cardioembolic stroke were not significantly different for any marker test. Eight patients taking aspirin at the time of the stroke had significantly lower 11-dTXB2 values than patients not taking aspirin (964 vs. 4,314 pg/mg of creatinine; P=0.007). Stroke patients not taking aspirin had significantly higher 11-dTXB2 concentration than age-matched controls (4,314 vs. 1,788 pg/mg of creatinine; P=0.006). Coagulation and platelet activation markers are increased in the acute phase of stroke regardless of the clinical mechanism. This finding suggests that the markers may not be useful for predicting clinical subtype of ischemic stroke in the acute phase.     

Analysis of proinflammatory and anti-inflammatory cytokine serum concentrations in patients with multiple sclerosis by using a multiplexed immunoassay

We examined cytokines and other inflammatory markers in serum samples from 833 patients with multiple sclerosis and 117 healthy control subjects. A multiplexed immunoassay was used to assess the concentrations of 13 cytokines/inflammatory markers: interferon (IFN)-γ; interleukins (ILs)-1β, 2, 4, 5, 6, 8, 10, 12, and 13; tumor necrosis factor (TNF)-α; IL-2 receptor; and soluble CD40 ligand. Significant increases between patients and control subjects were found for IFN-γ (mean, 7.5 vs 0.4 pg/mL; P = .0002), IL-2 (mean 5.7 vs 1.0 pg/mL; P =.0002), IL-1β (mean, 23.0 vs 11.3 pg/mL; P ≤ .0001), TNF-α (mean, 4.1 vs 1.2 pg/mL; P = .01), IL-4 (mean, 1.4 vs 0.1 pg/mL; P ≤ .0001), IL-10 (mean, 16.8 vs 7.5 pg/mL; P = .03), and IL-13 (mean, 4.5 vs 0.8 pg/mL; P ≤ .0001). Profiling cytokines in multiple sclerosis may help to identify mechanisms involved in the pathogenesis of the disease, aid in monitoring the disease course and in evaluating responses to specific therapies, and, potentially, lead to new therapies directed at cytokines or their receptors.     

Interaction between interleukin-6 and the natural anticoagulant system in acute stroke.

Inflammatory reactions mediated by cytokines are involved in the pathogenesis of acute stroke. Decrease in circulating levels of protein C (PC) and protein S (PS) induced by inflammatory cytokines has been postulated as a potential mechanism for a procoagulant tendency during acute stroke. The procoagulant state associated with impairments in natural anticoagulants may induce microvascular obstruction leading to a tissue perfusion reduction that worsens cerebral ischemia. Interleukin-6 (IL-6) regulate the synthesis of C4b-binding protein (C4BP), an acute-phase protein that also regulates PS plasma levels. We measured IL-6, C4BP, erythrocyte sedimentation rate (ESR), total and free PS and PC in 44 patients with acute ischemic stroke to determine if IL-6 decreases circulating levels of natural anticoagulants through the C4BP pathway and if these acute changes in natural anticoagulants may have clinical implications. Patients with higher levels of IL-6 had more severe neurologic deficits on admission, greater infarct size, higher levels of acute-phase reactants, and lower levels of free PS. IL-6 was significantly correlated with C4BP, ESR, and free PS levels. PC levels were also lower in the group of patients with greater IL-6, but differences were not statistically significant. No correlations were found between C4BP and natural anticoagulants. Severe neurologic deficit, greater infarct volume, atrial fibrillation, increased levels of inflammatory parameters (ESR and IL-6), and reduced levels of free PS were associated with disabling stroke at 3 months, but only neurologic severity and ESR remained as independent predictors of stroke disability on multiple regression analysis. Inflammatory reactions mediated by IL-6 during the acute phase of stroke influence the modulation of free PS. However, variations in free PS levels do not have implications for clinical outcome in stroke patients. The link between proinflammatory cytokines and free PS in the acute phase of stroke is not related to the C4BP pathway.     

Soluble interleukin-2 receptor and interleukin-4 in sera of asthmatic children before and after a prednisolone course.

BACKGROUND: Cytokine-mediated interactions among inflammatory cells may play a role in the pathogenesis of bronchial asthma. OBJECTIVE: To understand the role of soluble interleukin-2 receptor (sIL-2R) and interleukin-4 (IL-4) in the disease activity of acute asthma, changes in serum concentrations of sIL-2R and IL-4 elaborated by activated T-lymphocyte before and after prednisolone therapy with clinical improvement were determined in the present study. METHODS: Circulating levels of sIL-2R and IL-4 in sera from 15 normal control subjects and in sera from 20 allergic asthmatic children with acute exacerbation and in a stable condition were determined by using commercially available ELISA kits. RESULTS: The mean concentration of serum sIL-2R was significantly higher in acute exacerbation than in children with stable asthma (368.9 +/- 395.4 pg/mL vs 291.2 +/- 361.0 pg/mL; P < .01) or in control subjects (124.6 +/- 17.8 pg/mL; P < .001). The mean concentration of serum IL-4 was higher in acute exacerbation (5.82 +/- 1.10 pg/mL) and in stable asthmatic patients (6.73 +/- 2.83 pg/mL) versus control group subjects (5.54 +/- 1.20 pg/mL). However, the difference was not statistically significant among the three study groups. CONCLUSIONS: This study provides further evidence that changes in serum IL-2R may serve as an objective indicator for clinical outcome of allergic asthmatic patients.     

Thrombopoietin and interleukin-6 levels in Henoch-Sch?nlein purpura.

BACKGROUND AND PURPOSE: Depending on the severity of the illness, thrombocytosis is found in about 60% to 70% of patients with Henoch-Sch?nlein purpura (HSP). Whether thrombocytosis is the result of an inflammatory reaction mediated by thrombopoietin (TPO) or other inflammatory cytokines such as interleukin (IL)-6 remains unknown. METHODS: Thirty two patients who met the diagnostic criteria for HSP were included. They were divided into two groups - HSP patients with thrombocytosis (n = 14) and those without thrombocytosis (n = 18) with a platelet count of 400,000/microL. Eight normal healthy controls were also included. TPO and IL-6 serum levels during the acute phase were measured by enzyme-linked immunosorbent assay. RESULTS: Patients with platelet counts greater than 400,000/microL in the acute stage had significantly lower TPO levels than patients with platelet counts lower than 400,000/microL (310 +/- 65.6 pg/mL vs 608 +/- 97.8 pg/mL, p=0.013). However, HSP patients with or without thrombocytosis had similar TPO levels as the healthy controls (441 +/- 176 pg/mL, p=0.89 and 0.29, respectively). IL-6 serum levels were significantly elevated in HSP patients during the acute stage of HSP (28.6 +/- 61.7 pg/mL vs 3.16 +/- 1.35 pg/mL, p=0.049). In patients with complications of glomerulonephritis or gastrointestinal hemorrhage (n = 12), IL-6 levels were significantly lower than in those without such complications (8.07 +/- 3.79 pg/mL vs 40.9 +/- 16.9 pg/mL, p=0.007). CONCLUSIONS: This study showed that thrombocytosis in HSP patients is a type of inflammatory reactive thrombocytosis, and that IL-6 may also play a role in the pathogenesis of HSP.     

血清CXCL12和IL-33在急性缺血性脑卒中预后评估中的价值

目的:探讨血清CXCL12 和IL-33 在急性缺血性脑卒中预后评估中的价值。方法:选取2014 年12 月至2016年6 月到本院就诊的急性缺血性脑卒中患者作为病例组(122 例),同时选取本院同期健康体检人群作为正常组(59 名),采用ELISA 法检测研究对象入组时血清CXCL12 和IL-33 的水平。参照mRS 评分将患者分为预后良好组(86 例)和预后不良组(36 例),并通过受试者工作特征曲线(ROC)和曲线下面积(AUC)评价血清CXCL12 和IL-33 在急性缺血性脑卒中患者预后评估中的作用。结果:急性缺血性脑卒中患者血清CXCL12 水平明显高于对照组[8.0 ng/ ml(IQR,6.7-8.9) VS 3.0 ng/ ml(IQR,2.3-3.8),P<0.001],IL-33 水平明显高于对照组[65.25 ng/ L(IQR,56.05-71.08)VS 35.30 ng/ L(IQR,26.73-42.55),P<0.001];急性缺血性脑卒中预后良好组患者血清CXCL12 水平明显低于预后不良组[7.4 ng/ ml(IQR,6.3-8.3) VS 9.3 ng/mL(IQR,8.3-11.1),P<0.001],预后良好组患者IL-33 水平明显高于预后不良组[66.81 ng/ L(IQR,61.12-73.29)VS.55.38ng/ L(IQR,46.75-64.71),P<0.001]经Pearson 相关性分析显示血清CXCL12 和IL-33 水平与mRS 评分分别呈正相关和负相关(r =0.524,P<0.001;r =-0.443,P<0.001)。血清CXCL12、IL-33 评估急性缺血性脑卒中预后的曲线下面积分别为0.835、0.784,灵敏度分别为77.8%、83.4%,特异度分别为73.3%、66.7%。结论:血清CXCL12 和IL-33 可能作为急性缺血性脑卒中患者预后评估的标志物。     

Acute changes of coagulation and fibrinolysis parameters after experimental thromboembolic stroke and thrombolytic therapy

Thrombolysis is the only effective pharmaceutical therapy in acute ischemic stroke in humans but has a high risk of intracerebral hemorrhage. We aimed to establish an animal model to study changes of coagulation and fibrinolytic parameters during thromboembolic ischemic stroke and thrombolysis with recombinant tissue plasminogen activator (rt-PA). We used a thromboembolic stroke model in the rat. Animals were treated with rt-PA thrombolysis (n=10) and compared with untreated (n=10), sham operated (n=10) and control animals (n=20). Coagulation parameters (APTT, PT, TT, fibrinogen, AT III, TAT) and fibrinolytic parameters (t-PA antigen concentration, t-PA activity, PAI-1 concentration, PAI activity, plasminogen, antiplasmin) were measured at two time points (2.5 and 5h after stroke induction) with a battery of commercially available test kits. We observed an (1) initiation of coagulation and inhibition of fibrinolysis by the operation procedure itself, (2) simultaneous activation of fibrinolysis and its inhibitors after stroke induction and (3) potent initiation of fibrinolysis and consumption of fibrinolysis inhibitors after rt-PA therapy of stroke. We established a model system to monitor coagulation and fibrinolysis during thrombolytic therapy of stroke in the rat. This model may be used to study the influence of these parameters on hemorrhagic stroke transformation and outcome in experimental stroke in future.     

No increase in cervicovaginal proinflammatory cytokines after Carraguard use in a placebo-controlled randomized clinical trial

BACKGROUND: Assessment of cervicovaginal cytokine levels may be helpful to evaluate subclinical epithelial inflammation during safety evaluations of candidate microbicides. METHODS: Fifty-five HIV-seronegative Thai women were enrolled in a safety trial of the candidate microbicide Carraguard and were randomized to use Carraguard or placebo gel before vaginal sex. Cervicovaginal lavages were collected at baseline and after 1 month of gel use; levels of interleukin (IL)-1beta, IL-6, IL-8, and secretory leukocyte protease inhibitor (SLPI) were measured using microwell plate-based enzyme immunoassays. Median levels were compared between the baseline and 1-month follow-up visits using paired t tests; the median change between groups was compared using Wilcoxon rank sum tests. Women were examined for the presence of genital findings; the association between genital findings and cytokine levels was studied. RESULTS: No increase in levels of proinflammatory cytokines after use of Carraguard gel or placebo gel was observed during the study. The median change from the baseline to 1 month of follow-up was not significantly different between Carraguard and placebo groups (IL-1beta: -0.3 pg/mL vs. -3.93 pg/mL; P = 0.4, IL-6: -0.3 pg/mL vs. 0 pg/mL; P = 0.3, IL-8: -40.1 pg/mL vs. -53.2 pg/mL; P = 0.8, and SLPI: -26.5 pg/mL vs. 12.6 pg/mL; P = 0.07). Genital findings with intact epithelium were found in 16 (29%) women; these women tended to have somewhat higher IL-6 levels than those with normal epithelium (14.9 pg/mL vs. 8.8 pg/mL; P = 0.08). CONCLUSION: We found no increase in proinflammatory cytokines after Carraguard and placebo gel use, suggesting that neither gel causes inflammation. Further studies to assess the role of cytokines in microbicide safety studies are warranted.     

缺血性脑卒中发病早期血小板膜糖蛋白Ⅵ和血清白介素-6水平变化

目的:探讨血小板膜糖蛋白Ⅵ(GPVI)和白介素-6(IL-6)在急性缺血性脑卒中早期诊断中的意义。方法:分别采用流式细胞术和ELISA检测缺血性脑卒中患者和健康对照人群GPVI和IL-6水平。通过受试者工作曲线评估GPVI和IL-6的灵敏度、特异度及界值。结果:缺血性脑卒中病人血小板GPVI和血清IL-6水平显著高于对照组,差异有非常显著性(P0.01)。缺血性脑卒中病人GPVI与IL-6水平呈正相关(P0.01)。缺血性脑卒中病人GPVI的ROC曲线下面积为0.984,在最适诊断参考值17.76荧光强度(FI)时,诊断的灵敏度为90.00%,特异度为93.33%。IL-6的ROC曲线下面积为0.918,在最适诊断参考值0.55ng/L时,诊断的灵敏度为66.67%,特异度为86.67%。结论:血小板GPVI含量和血清IL-6水平升高与缺血性脑卒中有关。GPVI对缺血性脑卒中具有较高的诊断价值和准确度。     

Fibrinolytic changes during acute vascular damage induced by Mediterranean spotted fever

Hemostatic abnormalities and microvascular injury have been described in patients with Mediterranean spotted fever (MSF). Rickettsial vasculitis comprises endothelial injury and the immune and phagocytic host response. Only indirect evidence of activation of the fibrinolytic system has been reported in MSF, but no data on plasma levels of their main components, tissue type plasminogen activator (t-PA) and plasminogen activator inhibitor type 1 (PAI-1), in this disease are available. To obtain quantitative information on the ‘in vivo’ activation of the fibrinolytic system during the acute and subacute phase of the endothelial damage, several fibrinolytic components were studied in 28 MSF patients. Upon admission (day 1), patients showed a significant increase both in t-PA (30±20 ng/ml) and in PAI-1 antigen levels (55±30 ng/ml) as compared with normal levels (9±4 ng/ml and 11±4 ng/ml, respectively). The levels of PAI-1 activity were also increased (median: 7U/ml, range: 0–24 U/ml), but not significantly (normal values, median: 3 U/ml, range: 0.5–5 U/ml). After remission of the disease (day 30) a significant decrease in t-PA and PAI-1 antigen levels was observed in comparison with day 1, but the PAI-1 antigen concentration remained increased in comparison with normal values. The decrease in the ratio VIII:C/vWF:Ag found in the acute phase of the disease and after 30 days could indicate that the endothelial damage remains after this period. Tissue necrosis factor α (TNFα), a potential modulator of fibrinolytic system during gram-negative sepsis, showed normal values during the acute phase of the disease and at day 30. The data found in this study indicate that the changes observed in the fibrinolytic system during MSF could be due to the endothelial damage found in the acute phase of the disease. The increased plasma PAI-1 levels could contribute to an increase in the risk of venous thrombosis in MSF patients.     

Changes in serum and ascitic monocyte chemotactic protein-1 (MCP-1) and IL-10 levels in cirrhotic patients with spontaneous bacterial peritonitis.

Spontaneous bacterial peritonitis (SBP) is a prototypical infectious disease of cirrhotic patients. It has been suggested that cirrhotic patients' response to infection is less effective because of differences in the inflammatory and immune reactions. This study aimed to investigate the expression of the inflammatory cytokines monocyte chemotactic protein-1 (MCP-1) and interleukin-10 (IL-10) in cirrhotic patients with SBP. The MCP1 and IL-10 levels in the sera and ascitic fluids of cirrhotic patients with (n = 40) or without SBP (n=17) were serially analyzed by ELISA. In the non-SBP group, the mean MCP-1 levels in sera and ascites were 53.0 +/- 45.8 pg/mL and 197.5 +/- 109.5 pg/mL, respectively, and the IL-10 levels were 10.9 +/- 9.5 pg/mL and 77.6 +/- 79.7 pg/mL, respectively. In the SBP group, the mean MCP-1 levels in serum and ascites before treatment were 164.7 +/- 126.4 pg/mL and 365.3 +/- 583.0 pg/mL, respectively, and the IL-10 levels were 31.4 +/- 44.1 pg/mL and 188.1 +/- 189.5 pg/mL, respectively. The sera MCP-1 and ascites IL-10 levels differed significantly between the two groups. In the SBP group, sera and ascitic MCP-1 and IL-10 levels fell during treatment. The low MCP-1 and IL-10 levels on the seventh day of treatment were found to have a statistically significant relationship to patient survival. MCP-1 and IL-10 levels in sera and ascites may be related to the clinical course of SBP.     

In vitro fertilization. Do short-term changes in estrogen levels produce increased fibrinolysis?

High estrogen levels have been implicated as a cause of thrombosis. Patients undergoing in vitro fertilization (IVF) present an opportunity to study isolated estrogen effects on the coagulation system, since hormonal manipulation is used to achieve varying estrogen concentrations. We evaluated the effect of estrogen levels on the fibrinolytic and clotting systems of 32 patients undergoing IVF from whom venous samples were obtained during a period of several weeks at baseline and retrieval times. Flow cytometric studies were performed to assay platelet activation. Hemostasis markers measured thrombin-antithrombin complexes (TAT), fibrinogen, plasmin-alpha 2-antiplasmin complexes (PAP), plasminogen activator inhibitor (PAI-1), and D-dimer. A significant decrease in PAI-1 and an increase in PAP were observed at high estrogen levels, suggesting an increase in fibrinolytic potential. However, the products of fibrinolysis, as measured by D-dimer levels, decreased at high estrogen levels. No changes were seen in platelet activation indicators or TAT levels. The fact that a decrease in products of fibrinolysis was seen at high estrogen levels despite concurrent changes in PAI-1 and PAP that normally accompany increased fibrinolysis may be due to alterations in the overall balance of the fibrinolytic system or to a delay in onset of fibrinolysis.     

Association between platelet activation and fibrinolysis in acute stroke patients

We aimed to evaluate platelet activation and fibrinolyis in acute atherosclerotic ischemic stroke patients to clarify the relationship between them. Plasma P-selectin antigen, tissue plasminogen activator (tPA) antigen and activity, and plasminogen activator inhibitor-1 (PAI-1) activity were determined in 60 acute atherosclerotic stroke patients and matched control subjects. All patients were examined within 72 h after stroke onset. The levels of P-selectin, tPA antigen, and PAI-1 activity were all significantly higher in stroke patients compared with controls (all p < 0.0001); the level of tPA activity was significantly lower in patients than that in controls (p < 0.0001). These markers did not change much at different time points within 72 h. In stroke group, P-selectin concentration was highly correlated to PAI-1 activity (r = 0.8433, p < 0.001), but not to tPA antigen (r = -0.1752, p > 0.05), and tPA activity (r = 0.2465, p>0.05), which was further confirmed in the multiple linear regression analysis (F = 47.052, p < 0.0001). Our results indicate increased platelet activation and decreased fibrinolysis in patients with acute atherosclerotic ischemic stroke. Increased platelet activation may be correlated with decreased fibrinolysis.     

Extracellular matrix disturbances in acute myocardial infarction: relation between disease severity and matrix metalloproteinase-1, and effects of magnesium pretreatment on reperfusion injury.

PURPOSE: The purpose of this paper is to clarify the relationship between cytokines, matrix metalloproteinase-1 (MMP-1) and severity of acute myocardial infarction (AMI). Additionally, to investigate whether magnesium (Mg) sulfate pretreatment inhibits myocardial damage in coronary reperfusion therapy for patients with AMI. SUBJECTS: At first, 34 patients with AMI were enrolled. Then, the patients were classified into 2 groups with or without congestive heart failure (CHF) (C group and NC group, respectively). Interleukin 6 (IL-6), MMP-1 and the hemodynamic parameters were measured. Second, 36 AMI patients treated with coronary reperfusion therapy were enrolled. Patients were divided into 2 groups (18 patients each) as the non-pretreated group (Control group) and the group pretreated with intravenous Mg sulfate (0.27 mmol/kg) (Mg group). IL-6, MMP-1 and the indexes of reperfusion injury were evaluated. RESULTS: There were positive correlations between peak MMP-1 level, and peak creatine kinase value and pulmonary capillary wedge pressure and peak IL-6 level (r = 0.43, r = 0.70, and r = 0.60, respectively) in all patients. There were negative correlations between peak MMP-1 level and left ventricular ejection fraction and cardiac index (r = - 0.52 and r = - 0.55, respectively). The peak blood IL-6 and MMP-1 level increased in AMI, particularly in patients with CHF (C group vs NC group; 130 vs 51 pg/mL and 37 vs 18 ng/mL, both p < 0.01). Additionally, peak IL-6 and peak MMP-1 in the Mg group were lower than those of the control group (39 vs 92 pg/mL and 16 vs 20 ng/mL, p < 0.05 and p = 0.09, respectively). The incidence of reperfusion injury including reperfusion arrhythmia and transient exacerbation of ST elevation in the Mg group was lower than that of control group (17 vs 78% and 2.5 vs 4.7 mm, p < 0.01 and p = 0.08, respectively). CONCLUSION: These results may suggest that the severity of AMI is reflected by the blood IL-6 and MMP-1 levels and that pretreatment with Mg administration protects the myocardium of patients with AMI from reperfusion injury induced by IL-6 and MMP-1.     

Inflammatory mediators of systemic inflammation in neonatal sepsis

Objective and design

Sepsis refers to severe systemic inflammation in response to invading pathogens. To understand the molecular events that initiate the systemic inflammatory response, various inflammatory mediators were analyzed in neonatal sepsis samples and compared with normal samples.

Materials and methods

We initially measured the levels of the various classical inflammatory mediators such as acute phase proteins [C-reactive protein (CRP) and procalcitonin (PCT)], granule-associated mediators (NE, MPO and NO), proinflammatory cytokines [tumour necrosis factor-α (TNFα), IL-1β and IL-6), antiinflammatory cytokines (IL-10 and IL-13) and chemokines [IL-8 and monocyte chemotactic protein (MCP-1)] and novel cytokines (IL-12/IL-23p40, IL-21 and IL-23) using ELISA. We also used the human inflammation antibody array membrane to profile the inflammatory proteins that are involved in neonatal sepsis.

Results

There were significantly higher levels of CRP (5.4 ± 0.70 mg/L), PCT (1.500 ± 0.2400 μg/L); NE (499.2 ± 22.01 μg/L), NO (54.22 ± 3.131 μM/L); TNFα (396.6 ± 37.40 pg/mL), IL-1β (445.3 ± 34.25 pg/mL), IL-6 (320.9 ± 43.38 pg/mL); IL-8 (429.5 ± 64.08 pg/mL) MCP-1 (626.25 ± 88.91 pg/mL), IL-10 (81.80 ± 9.45 pg/mL), IL-12/IL-23p40 (30.25 ± 0.6 pg/mL), IL-21 (8,263.3 ± 526.8 pg/mL) and IL-23 (6,083 ± 781.3 pg/mL) in neonates with sepsis compared to normal. The levels of MPO (21.20 ± 3.099 ng/mL) were downregulated, whereas there was no change in IL-13 (188.7 ± 10.63 pg/mL) levels in septic neonates when compared with normal. Using the human inflammation antibody array membrane, we detected the presence of 17 inflammatory proteins such as IL-3, IL6R, IL12p40, IL-16, TNFα, TNFβ, TNF R1, chemokines I-309, IP-10 (IFN-γ inducible protein 10), MCP-1, MCP-2, MIP 1β (macrophage inflammatory protein), MIP-1δ, eotaxin-2, growth factors TGFβ1 (transforming growth factor beta), PDGF (platelet derived growth factor), and cell adhesion molecule ICAM-1 (intracellular adhesion molecule) that were upregulated whereas RANTES which was downregulated in neonatal sepsis.

Conclusion

The simultaneous secretion and release of multiple mediators such as proinflammatory cytokines and chemokines, cell adhesion molecules, and growth factors were found to be involved in the initiation of systemic inflammation in neonatal sepsis. Therefore, measuring the concentration of multiple mediators may help in the early detection of neonatal sepsis and help to avoid unnecessary antibiotic treatment.     

Early induction of IL-1 receptor antagonist (IL-1Ra) in infants and children undergoing surgery.

The cytokine response to injury or trauma is of interest in terms of both its mediation of the acute phase response and its possible relation to the immunological depression observed after major surgery. In this study, the production of cytokines IL-1 beta, tumour necrosis factor-alpha (TNF-alpha), IL-6 and the naturally occurring inhibitor of IL-1, IL-1Ra, have been investigated in infants and children undergoing Swenson's pull-through operation for Hirschsprung's disease. Samples of peripheral blood were taken before, during and after surgery for the measurement of cytokines. IL-1Ra levels increased significantly (P < 0.01) at 2 h after commencement of surgery, with maximal levels for individual patients being attained between 3 h and 5 h (range 7.6-67.9 ng/ml). The mean level of IL-1Ra was maximal (26.2 ng/ml) at 5 h and returned to baseline levels between 24 h and 72 h. There were no changes observed in the circulating levels of IL-1 beta in nine out of 11 patients following commencement of surgery. TNF-alpha levels did not increase in any of the patients studied. IL-6 levels increased significantly (P < 0.02) 3 h after commencement of surgery, reaching maximum concentrations at 24 h (range 20-670 pg/ml), with levels falling between 48 h and 72 h. This study demonstrates, in vivo, the independent induction of IL-1Ra without a concomitant increase of IL-1 beta levels after major surgery. It also shows that IL-1Ra is the earliest cytokine produced in response to surgical stress.     

Induction of phagocyte-stimulating cytokines by in vitro stimulation of human peripheral blood mononuclear cells with Haemophilus influenzae

The aim of this study was to analyse the in vitro response of human peripheral blood mononuclear cells to stimulation with killed Haemophilus influenzae strains of different capsular types, isolation sites and from cases with different forms of infections. The mean stimulatory index using 10(6) bacteria/well was 10, and 80 when 10(8) bacteria/well were used for stimulation. The mean+/-SD level was 13+/-4 ng/ml for interleukin (IL)-1beta, 128+/-73 ng/ml for IL-6, 203+/-122 ng/ml for IL-8, 3160+/-1220 pg/ml for IL-10, 29+/-40 pg/ml for IL-12, 2800+/-1790 pg/ml for tumour necrosis factor (TNF)-alpha and 4+/-7 ng/ml for interferon (IFN)-gamma, when stimulating cells with the lower dose of 10(6) bacteria/well. Using the higher bacterial dose, the levels of IL-1beta, TNF-alpha and IL-12 remained similar, whereas the IL-6, IL-8 and IL-10 levels were significantly lower, and IFN-gamma levels were significantly higher. Strains isolated from the bronchial tree induced significantly higher levels of IFN-gamma and significantly lower levels of IL-6, IL-8 and IL-10 than strains from other isolation sites. In conclusion, H. influenzae generated phagocyte-activating cytokines and an IL-10/IL-12 ratio that was 1090 times that described previously for Streptococcus pneumoniae.     

Dose-dependent increase in plasma interleukin-6 after recombinant tumour necrosis factor infusion in humans.

Several studies have shown that the cytokine interleukin-6 (IL-6) is produced in response to tumour necrosis factor (TNF) in vitro. This study examines the in vivo relation between these two cytokines with assays of plasma IL-6 and TNF levels in subjects with chronic hepatitis B undergoing immunomodulatory therapy with recombinant TNF (rTNF). Plasma IL-6 was detected from 20 min after rTNF infusion with levels peaking after 2-3 h and levels correlated with the dose of rTNF administered (r = 0.67, P = 0.004). Peak levels of IL-6 (mean 295, range 266-297 ng/l) were lower than those seen in certain disease states despite the very high peak levels of rTNF (mean 11,750, range 5623-18,620 ng/l). These findings suggest that the very high levels of IL-6 found in certain disease states are not purely the result of circulating TNF. Other factors such as endotoxin or other cytokines may also play a role in determining levels of plasma IL-6.