Interleukin 12 induces activation of fibrinolysis and coagulation in humans

Interleukin 12 (IL-12) has potential efficacy in malignant, infectious and allergic diseases. Its side-effects include activation of coagulation and fibrinolysis, as documented in chimpanzees. We assessed the coagulative and fibrinolytic response in 18 patients with renal cell carcinoma after subcutaneous injection of 0.5 microg/kg recombinant human IL-12. IL-12 induced a fibrinolytic response in 17 patients (94%): plasmin-alpha2-anti-plasmin complexes (PAPc) increased from 11.8 +/- 6.6 nmol/l (mean +/- SD) to a maximum of 18.8 +/- 7.4 nmol/l at 72 h. Baseline levels of tissue plasminogen activator (tPA) and plasminogen-activator inhibitor-I (PAI) were elevated in eight and 14 patients respectively. tPA increased from 12.6 +/- 5.2 ng/ml to a maximum of 19.0 +/- 6.7 ng/ml at 72 h. PAI decreased from 111 +/- 69 ng/ml to a minimum of 65 +/- 53 ng/ml at 8 h, thereafter remaining below baseline. Elevation of PAPc correlated with elevation of tPA and reduction of PAI. A coagulative response occurred in nine patients (50%): thrombin-anti-thrombin III complexes increased from 29 +/- 53 ng/ml to a maximum of 460 +/- 322 ng/ml at 12 h. Patients with and without a coagulative response had similar levels of recombinant human IL-12, interferon-gamma or tumour necrosis factor-alpha. We conclude that IL-12 can activate both fibrinolysis and coagulation in a significant proportion of patients with cancer. The time-frame and sequence of these activation processes differ from those known for other cytokines.     

Blood coagulation activation and fibrinolysis during a downhill marathon run.

Prolonged physical exercise is associated with multiple changes in blood hemostasis. Eccentric muscle activation induces microtrauma of skeletal muscles, inducing an inflammatory response. Since there is a link between inflammation and coagulation we speculated that downhill running strongly activates the coagulation system. Thirteen volunteers participated in the Tyrolean Speed Marathon (42,195 m downhill race, 795 m vertical distance). Venous blood was collected 3 days (T1) and 3 h (T2) before the run, within 30 min after finishing (T3) and 1 day thereafter (T4). We measured the following key parameters: creatine kinase, myoglobin, thrombin-antithrombin complex, prothrombin fragment F1 + 2, D-dimer, plasmin-alpha(2)-antiplasmin complexes, tissue-type plasminogen activator antigen, plasminogen-activator-inhibitor-1 antigen and thrombelastography with ROTEM [intrinsic pathway (InTEM) clotting time, clot formation time, maximum clot firmness, alpha angle]. Thrombin generation was evaluated by the Thrombin Dynamic Test and the Technothrombin TGA test. Creatine kinase and myoglobin were elevated at T3 and further increased at T4. Thrombin-antithrombin complex, prothrombin fragment F1 + 2, D-dimer, plasmin-alpha(2)-antiplasmin complexes, tissue-type plasminogen activator antigen and plasminogen-activator-inhibitor-1 antigen were significantly increased at T3. ROTEM analysis exhibited a shortening of InTEM clotting time and clot formation time after the marathon, and an increase in InTEM maximum clot firmness and alpha angle. Changes in TGA were indicative for thrombin generation after the marathon. We demonstrated that a downhill marathon induces an activation of coagulation, as measured by specific parameters for coagulation, ROTEM and thrombin generation assays. These changes were paralleled by an activation of fibrinolysis indicating a preserved hemostatic balance.     

CO2 artificial pneumothorax on coagulation and fibrinolysis during thoracoscopic esophagectomy

Background:CO₂ artificial pneumothorax creates a sufficient operative field for thoracoscopic esophagectomy. However, it has potential complications and continuous CO₂ insufflation may impede coagulation and fibrinolysis. We sought to compare the effects of CO₂ artificial pneumothorax on perioperative coagulation and fibrinolysis during thoracoscopic esophagectomy.Methods:We investigated patients who underwent thoracoscopic esophagectomy with (group P, n = 24) or without CO₂ artificial pneumothorax (group N, n = 24). The following parameters of coagulation–fibrinolysis function: intraoperative bleeding volume; serum levels of tissue plasminogen activator (t-PA), plasminogen activator inhibitor (PAI-1), thromboelastogram (TEG), D-Dimer; and arterial blood gas levels were compared with two groups.Results:Group P showed higher levels of PaCO₂, reaction time (R) value and kinetics (K) value, but significantly lower pH value, alpha (α) angle and Maximum Amplitude (MA) value at 60 minutes after the initiation of CO₂ artificial pneumothorax than group N ((P < .05, all). The t-PA level after CO₂ insufflation for 60 minutes was significantly higher in group P than in group N (P < .05), but preoperative levels were gradually restored on cessation of CO₂ insufflation for 30 min (P > .05). There was no significant difference in D-dimer.Conclusion:CO₂ artificial pneumothorax during thoracoscopic esophagectomy had a substantial impact on coagulation and fibrinolysis, inducing significant derangements in pH and PaCO₂.Trial registration:The study was registered at the Chinese clinical trial registry (ChiCTR1800019004)     

Tumor necrosis factor-alpha induces activation of coagulation and fibrinolysis in baboons through an exclusive effect on the p55 receptor

Tumor necrosis factor-alpha (TNF-alpha) can bind to two distinct transmembrane receptors, the p55 and p75 TNF receptors. We compared the capability of two mutant TNF proteins with exclusive affinity for the p55 or p75 TNF receptor with that of wild type TNF, to activate the hemostatic mechanism in baboons. Both activation of the coagulation system, monitored by the plasma levels of thrombin-antithrombin III complexes, and activation of the fibrinolytic system (plasma levels of tissue-type plasminogen activator, and plasminogen activator inhibitor type I), were of similar magnitude after intravenous injection of wild type TNF or the TNF mutant with affinity only for the p55 receptor. Likewise, wild type TNF and the TNF p55 specific mutant were equally potent in inducing neutrophil degranulation (plasma levels of elastase- alpha 1-antitrypsin complexes). Wild type TNF tended to be a more potent inducer of secretory phospholipase A2 release than the p55 specific TNF mutant. Administration of the TNF mutant binding only to the p75 receptor did not induce any of these responses. We conclude that TNF-Induced stimulation of coagulation, fibrinolysis, neutrophil degranulation, and release of secretory phospholipase A2 are predominantly mediated by the p55 TNF receptor.     

Interleukin-2 administration causes reversible hemodynamic changes and left ventricular dysfunction similar to those seen in septic shock

Interleukin-2, a lymphocyte product, has well demonstrated antitumor activity in humans. Early clinical studies showed hemodynamic alterations in patients receiving the drug as antitumor immunotherapy. We serially assessed interleukin-2-associated hemodynamic parameters and left ventricular ejection fractions in five patients with neoplastic diseases unresponsive to conventional therapies. By day 4 of therapy, compared with baseline (preinterleukin-2), all patients developed tachycardia (p less than 0.01), decreased mean arterial blood pressure (p less than 0.05), increased cardiac index (p less than 0.05), and decreased systemic vascular resistance (p less than 0.01). In addition, left ventricular ejection fraction fell from 58.0 +/- 4.7 to 36.4 +/- 4.0 percent (0.05 less than p less than 0.10), which was associated with a trend toward left ventricular dilatation manifested by an increase in left ventricular end-diastolic volume index. Transient renal dysfunction was noted in all five patients, and one developed transient respiratory failure; both types of organ dysfunction recovered to baseline values after cessation of immunotherapy. Thus, interleukin-2 induces multiple reversible cardiovascular abnormalities that are similar to the hemodynamic manifestations of human septic shock.     

Markers of activation of coagulation and fibrinolysis in patients with Cushing's syndrome

Patients with active Cushing's syndrome have an increased thrombotic tendency. We chose to reassess the mechanism underlying the thrombophilic state associated with this clinical condition using sensitive markers of coagulation and fibrinolysis activation in 17 patients with active disease. The results were compared with those obtained in 12 Cushing's patients successfully treated by surgery and in 20 normal individuals. The general pattern of results in patients with active disease was the finding of increased levels of von Willebrand factor (VWF: Ag), a marker of enhanced metabolic function of endothelial cells (VWF:Ag 181 +/- 42 vs 110 +/- 43, p<0.001 in normal subjects), accompanied by signs of heightened thrombin and plasmin generation, expressed by high levels of thrombin-antithrombin (TAT 5.59+/-3.6 vs 3.06+/-0.92 ng/ml in controls, p<0.01) and plasmin-antiplasmin complexes (PAP 407+/-176 vs 245+/-67 ng/ml in controls, p<0.01). VWF:Ag and TAT values were significantly higher in hypertensive than in normotensive patients with active disease (205+/-40 vs 155+/-26 U/dl, p<0.05 and 7.49+/-3.7 vs 3.45+/-1.8, p<0.01, respectively). Plasma levels of plasminogen activator inhibitor type 1 were higher, though not to a statistically significant extent, in patients with active disease compared to controls (12.8+/-12.3 vs 5.6+/-7.4 IU/ml, NS) and positively correlated with body mass index (r=0.66, p<0.01). After surgical control of Cushing's syndrome, there was a partial or complete reversal of the abnormalities to values similar to those found in normal individuals. Our data suggest that the thrombophilic state present in patients with active Cushing's syndrome is related to an enhanced metabolic function of endothelial cells; this in turn may be caused by an heightened production of thrombin with secondary hyperfibrinolysis. Primary prophylaxis with anticoagulants is recommended in these patients when they are exposed to a thrombophilic condition such as surgery.     

Interleukin-6 changes deformability of neutrophils and induces their sequestration in the lung

Interleukin-6 (IL-6) is an important mediator of both the hepatic and the bone marrow components of the acute-phase response. Previous studies from our laboratory have shown that cells released into the circulation from the marrow preferentially sequester in the lung. The present study was designed to examine the mechanism of this sequestration using a single dose of recombinant human IL-6 to stimulate the marrow in rabbits. Marrow release was monitored by labeling polymorphonuclear leukocyte (PMN) precursors in the marrow with the thymidine analogue, 5'-bromo-2-deoxyuridine (BrdU), 24 h before IL-6 treatment. This treatment caused a neutrophilia that was associated with the increase of circulating BrdU- labeled PMN (PMN(BrdU)) and morphometric studies confirmed that PMN(BrdU) released from the marrow preferentially sequestered in the lung microvessels compared to unlabeled PMN. IL-6 treatment increases PMN F-actin content (p < 0.05) that was not due to cell activation by IL-6. In vitro studies show that IL-6 treatment decreased the deformability of circulating PMN (p < 0.05). These studies confirm that IL-6 treatment causes an accelerated release of PMN from the bone marrow and shows that these newly released PMN have high levels of F-actin, are less deformable, and preferentially sequester in lung microvessels.     

Inhibition of coagulation,fibrinolysis, and endothelial cell activation by a p38 mitogen-activated protein kinase inhibitor during human endotoxemia

P38 mitogen-activated protein kinase (MAPK) is an important component of intracellular signaling cascades that initiate various inflammatory cellular responses. To determine the role of p38 MAPK in the procoagulant response to lipopolysaccharide (LPS), 24 healthy subjects were exposed to an intravenous dose of LPS (4 ng/kg), preceded 3 hours earlier by orally administered 600 or 50 mg BIRB 796 BS (a specific p38 MAPK inhibitor), or placebo. The 600-mg dose of BIRB 796 BS strongly inhibited LPS-induced coagulation activation, as measured by plasma concentrations of the prothrombin fragment F1 + 2. BIRB 796 BS also dose dependently attenuated the activation and subsequent inhibition of the fibrinolytic system (plasma tissue-type plasminogen activator, plasmin-alpha2-antiplasmin complexes, and plasminogen activator inhibitor type 1) and endothelial cell activation (plasma soluble E-selectin and von Willebrand factor). Activation of p38 MAPK plays an important role in the procoagulant and endothelial cell response after in vivo exposure to LPS.     

Histidine-rich glycoprotein: a possible modulator of coagulation and fibrinolysis

Histidine-rich glycoprotein (HRG) is one of the major plasma proteins and thought to function in blood coagulation, fibrinolysis, and innate immune systems. The amino acid sequence of HRG revealed a multidomain structure consisting of cystatin-like domains 1 and 2, a Pro-rich domain 1, a His-rich domain, a Pro-rich domain 2, and a C-terminal domain. Broad ligand-binding properties of HRG are involved in the multivalent functions of HRG. Among various functions of HRG, its interactions with heparin/heparan sulfate, fibrinogen, and plasminogen are thought to be intimately related to its roles in the coagulation and fibrinolytic systems. Recent studies on these topics are mainly reviewed in this article. The newly disclosed abilities of HRG in angiogenesis, its antibacterial effect, its activation of T-cell lines in cooperation with Concanavalin A, and the identification of a putative receptor for HRG on T cell lines are also described.     

Exploration of rapid bedside monitoring of coagulation and fibrinolysis parameters during thrombolytic therapy.

Monitoring coagulation parameters during thrombolytic therapy could be useful for prediction and treatment of haemorrhagic episodes. Technology based on dry reagent chemistry has been developed that allows rapid (less than 10 min) assays on small samples of whole blood. The assay principle is based on the restriction of motion of paramagnetic particles during fibrin polymerization, and subsequent liberation of particle motion during fibrinolysis. This technology was used to monitor prothrombin time (PT), activated partial thromboplastin time (aPTT), fibrinogen levels and fibrinolysis profiles during thrombolytic therapy with tissue plasminogen activator for acute myocardial infarction. The PT and aPTT obtained with the COAG-1 correlated well with conventional assays (r = 0.93 and 0.92 for PT and aPTT, respectively; p = 0.0001). Fibrinogen estimates, obtained by COAG-2 also correlated well with modified Clauss assays (r = 0.86, p = 0.0001). The rapid determination of the aPTT may improve management of adjunctive anticoagulant therapy following thrombolysis. The fibrinolysis profile may be useful during thrombolytic therapy to verify that a lytic state has been achieved, to monitor the lytic state throughout therapy, and to verify that the lytic state normalizes once therapy has been completed.     

Studies on oral contraceptive-induced changes in blood coagulation and fibrinolysis and the estrogen effect on endothelial cells

Summary Blood coagulation (fibrinogen, thrombinantithrombin III complexes, TAT, and prothrombin fragment F₁₊₂) and fibrinolytic parameters [fibrin split-product D-dimer, tissue plasminogen activator (t-PA) activity, plasminogen activator inhibitor-1 activity (PAI-1), and plasmin-antiplasmin-complexes (PAP)] were evaluated in 16 women on low estrogen (EE) oral contraceptive (OC) therapy. Blood samples were taken before and between days 18 and 22 of the first, third, and sixth treatment cycle. Fibrinogen levels were found significantly elevated during OC treatment compared with pretreatment values, while TAT and also F₁₊₂ levels remained unchanged. Treatment-induced activation of fibrinolysis was documented by elevated D-dimer [pretreatment (pt): 172 ng/ml (range: 65–640 ng/ml), cycle 6 (c.6): 351 ng/ml (range: 93–960 ng/ ml),p<0.05)] and PAP [(pt: 46.6 ng/ml (13–220 ng/ml), c.6: 66.4 ng/ml (21–200 ng/ml),p<0.05] plasma levels. Among the fibrinolytic components a decrease in PAI-1 [pt: 10.8 ng/ml (2–56 ng/ml), c.6: 5.3 ng/ml (2.2–14.4 ng/ml),p<0.05] and an increase in t-PA activity [pt: 0.23 U/ml (0.17–0.45 U/ml), c.6: 0.33 U/ml (0.2–0.9 U/ml),p<0.05] were detected. Experiments with cultured human endothelial cells (EC) showed that EE influenced neither EC hemostatic regulatory activities (tissue factor, thrombomodulin) nor the secretion of the fibrinolytic components t-PA and PAI-1.     

Interleukin-1 induces lipid peroxidation and membrane changes in rat hippocampus: An age-related study

BACKGROUND: The proinflammatory cytokine, interleukin-1, is traditionally associated with the immune response but recent evidence indicates that it plays a role in neuronal function. Its expression is increased in neurodegenerative conditions and preliminary evidence suggests that it is also increased with increasing age. Receptors for interleukin-1 are differentially distributed in the brain with a high density in the hippocampus, where interleukin-1beta exerts inhibitory effects on release and calcium channel function. OBJECTIVE: The aim of this study was to investigate the possibility that interleukin-1 might lead to age-related changes in membrane composition. METHODS: Lipid peroxidation was assessed in the presence or absence of interleukin-1beta in hippocampal tissue prepared from 4- and 22-month-old rats. These data were analysed in parallel with age-related changes in arachidonic acid and interleukin-1beta concentrations in the hippocampus. RESULTS: We report that interleukin-1beta increased lipid peroxidation in hippocampal tissue prepared from 4- but not 22-month-old rats, and that this effect was inhibited by alpha-tocopherol. The attenuated response to interleukin-1beta in tissue prepared from aged rats correlated with increased expression of endogenous interleukin-1beta. Thus, using an ELISA, we have demonstrated an age-related increase in the concentration of interleukin-1beta, which is accompanied by an age-related decrease in membrane arachidonic acid. CONCLUSION: We propose that increased interleukin-1beta expression impacts on membrane composition and therefore contributes to age-related impairments in neuronal function.     

二氧化碳气腹时间对老年腹腔镜胆囊切除术患者凝血-纤溶和血管内皮细胞活性的影响

目的 观察二氧化碳(CO2)气腹时间对老年人腹腔镜胆囊切除术(LC)患者凝血-纤溶和血管内皮细胞活性的影响.方法 胆石症择期行LC患者45例,年龄＞60岁,术后根据气腹持续时间分组:气腹时间≤60 min组21例;气腹时间＞60 min组24例.于入院检查时(术前)、术毕、术后第1、2、3天抽取静脉血检测凝血酶原时间(PT)、激活部分凝血活酶时间(APTT)、凝血酶原片段1+2(F1+2)浓度、抗凝血酶-Ⅲ(AT-Ⅲ)活性、纤维蛋白原(Fib)浓度、组织纤溶酶原激活物(t-PA)浓度、纤溶酶原激活物抑制物-1(PAI-1)浓度、D-二聚体(D-D)浓度、血管性血友病因子(vWF)活性.结果 (1)凝血指标:术后第3天,＞60 min组的,F1+2为 (1.60±0.26) μg/L,高于≤60 min组的(1.32±0.24) μg/L(P＜0.05);AT-Ⅲ为(84.82±20.21)%,低于≤60 min组的(97.49±16.87)%(P＜0.05);术后第2、3天的Fib分别为(3.87±0.62)、(3.98±0.77)g/L,高于≤60 min组的 (3.42±0.72)、(3.42±0.63)g/L(P＜0.05).(2)纤溶-抗纤溶指标:＞60 min组术后第2 、3天的PAI-1为(33.93±10.42)、(32.90±11.25) μg/L高于≤60 min组的(26.69±9.49)、(26.31±7.06)μg/L(P＜0.05).(3)血管内皮细胞活性指标:＞60 min组术后第2 、3天的vWF为(174.53±44.03)%、(176.31±47.6)%,高于≤60 min组的(134.37±37.74)%、(131.21±36.34)% (P＜0.05).结论 老年LC患者,术后有明显的凝血-纤溶激活和血管内膜损伤;随气腹时间延长,凝血激活和纤溶抑制程度高,凝血-纤溶相对不平衡,血管内膜损伤更明显,可能增加血栓形成风险.

Abstract：

Objective To observe the effect of duration of carbon dioxide pneumoperitoneum on coagulation, fibrinolysis and endothelial activation in elderly patients undergoing laparoscopic cholecystectomy (LC). Methods The 45 elderly patients with cholelithiasis scheduled for LC, aged over 60 yeas, were placed in different groups respectively after surgery according to the duration of pneumoperitoneum. The duration of pneumoperitoneum was ≤60 minutes in group A (n=21),and more than 60 minutes in group B (n=24). Venous blood samples were taken on admission (baseline), at the end of surgery, the 1st, 2nd and 3rd day after surgery for determination of prothrombin time (PT), activated partial thromboplastin time (APTT), prothrombin fragment F1+2 (F1+2), antithrombin 3 (AT-Ⅲ activity), fibrinogen (Fib), tissue plasminogen activator (t-PA), plasminogen activator inhibitor type-1 (PAI-1), D-dimer (D-D), von Willebrand factor (vWF activity). Results Concerning the coagulation activation, at the 3rd postoperative day, the level of F1+2 was significantly higher in group B than in group A [(1.60±0.26) μg/L vs. (1.32±0.24) μg/L, P＜0.05]; AT-III was significantly higher in group B than in group A [(84.82%±20.21%) vs. (97.49%±16.87%), P＜0.05]. At the 2nd and 3rd postoperative day, the levels of Fib were significantly higher in group B than in group A [(3.87±0.62) g/L vs. (3.42±0.72) g/L, (3.98±0.77) g/L vs. (3.42±0.63) g/L, respectively, P＜0.05]. Concerning fibrinolysis, But at the 2nd and 3rd postoperative day, the level of PAI-1 was significantly higher in group B than in group A [(33.93±10.42) μg/L vs. (26.69±9.49) μg/L, (32.90±11.25) μg/L vs. (26.31±7.06) μg/L respectively, P＜0.05]. Concerning endothelial activation, at the 2nd and 3rd postoperative day, vWF was significantly higher in group B than in group A [(174.53%±44.03%) vs. (134.37%±37.74%), (176.31%±47.6%) vs. (131.21%±36.34%), respectively, P＜0.05]. Conclusions Marked activations of coagulation-fibrinolysis and endothelial activation are observed postoperatively in elderly patients undergoing laparoscopic cholecystectomy. Along with prolonged duration of pneumoperitoneum, more pronounced alterations of increased coagulation, reduced fibrinolysis and endothelial activation are observed, which could constitute an imbalanced situation of coagulation-fibrinolysis and increases the risk of venous thrombosis.