Endotoxin, TNF-alpha, interleukin-6 and parameters of the cellular immune system in patients with intraabdominal sepsis.

The correlation of endotoxin (ET), tumor necrosis factor alpha (TNF-alpha), interleukin-6 (IL-6), and cellular immune parameters with multiple organ failure and lethal outcome in intraabdominal infections was studied in a group of 18 patients with peritonitis, abscess or pancreatitis. Of these patients, 7 developed respiratory failure and 5 died due to multiple septic organ failure. The peak levels of ET (2.7 +/- 1.3 ng/ml) in the course of the disease were followed by moderate increases of TNF-alpha (mean 147 +/- 41 pg/ml) and IL-6 (170 +/- 61 pg/ml) within 2 days. Analysis of the parameters for the last 12 days prior to death or discharge showed, that the patient group with lethal outcome was characterized by significant lower mean plasma levels of TNF-alpha (less than 75 pg/ml versus greater than 160 pg/ml) and IL-6 (less than 130 pg/ml versus greater than 270 pg/ml), as well as high rates of unstimulated thymidine uptake into peripheral mononuclear blood cells (greater than 44000 cpm/8 x 10(6) PMBC/18 h versus less than 24000 cmp), T-lymphocyte depression (CD3; approximately greater than 40% reduction) with lower T-helper/inducer subset cell numbers (mean CD:CD8 ratio 1.0 +/- 0.55 versus 1.8 +/- 0.2) and lower lectin (PHA) stimulation values (1.9 +/- 1.4 versus 4.1 +/- 1.0). These data demonstrate an anergic immune status with low mediator levels and depressed T-lymphocyte function in patients with poor prognosis.(ABSTRACT TRUNCATED AT 250 WORDS)     

鼠白细胞介素12质粒对小鼠支气管哮喘模型气道炎症及细胞因子的影响

目的　研究鼠白细胞介素 12 (IL 12 )基因表达 (mIL 12 )质粒对小鼠支气管哮喘 (简称哮喘 )模型气道炎症及细胞因子的影响 ,并分析其作用机制。方法　卵白蛋白 (OVA)致敏建立小鼠哮喘模型。BALB/c小鼠 4 1只 ,分为 6组。即哮喘模型组 (A组 ) 8只 :OVA致敏 +OVA气雾攻击 ;模型对照组 (B组 ) 6只 :用生理盐水代替 1%OVA溶液雾化 ,余处理同A组 ;mIL 12质粒预防组 (C组 ) 8只 :实验第 1、3、5天各肌肉注射mIL 12质粒 10 0 μg ;mIL 12质粒治疗组 (D组 ) 8只 :实验第 14、16、18天各肌肉注射mIL 12质粒 10 0 μg ;空质粒预防组 (E组 ) 5只 :实验第 1、3、5天各肌肉注射空质粒 10 0 μg ;空质粒治疗组 (F组 ) 6只 :实验第 14、16、18天各肌肉注射空质粒 10 0 μg。测定所有小鼠支气管 肺泡灌洗液 (BALF)中的嗜酸粒细胞 (EOS)个数和IL 4、IL 5、γ干扰素 (IFN γ)水平。结果B组小鼠EOS个数和IL 4、IL 5、IFN γ浓度分别为 (0 0 1± 0 0 3)× 10 8/L、(2 4± 4 ) pg/ml、(33± 6 ) pg/ml、(72 5± 5 9) pg/ml,C组为 (0 0 6± 0 0 4 )× 10 8/L、(43± 13) pg/ml、(6 3± 10 )pg/ml、(6 2 6± 6 0 ) pg/ml,D组为 (0 11± 0 12 )× 10 8/L、(38± 14 )pg/ml、(6 6± 14 ) pg/ml、(6 6 1± 4 0 ) pg/ml,与A     

Stimulating effect of growth hormone on cytokine release in children

OBJECTIVE: The aim of the present study was to investigate the effect of exogenously administered GH on serum levels of interleukin (IL)-1beta, IL-2, IL-12, tumor necrosis factor (TNF)-alpha and interferon (IFN)-gamma and their relation with IGF-I levels in normal short stature children. DESIGN AND METHODS: 23 short prepubertal non GH-deficient children (10 females and 13 males) whose mean+/-s.d. chronological age was 11.95+/-1.85 Years (from 8.80 to 14.89 Years), and mean+/-s.d. bone age was 10.48+/-2.44 Years, were evaluated during a somatomedin generation test (human GH 0.1 IU/kg per day for 4 days) to exclude a partial GH resistance as the cause of short stature; 34 sex- and age-matched healthy subjects were studied as controls. Circulating cytokine values were measured in basal conditions in all children, and 12 h following the 4th GH subcutaneous injection in the 23 short children only. RESULTS: No significant differences were found between short children and controls in basal values of serum IGF-I (192.1+/-18.3 and 198.2+/-28.2 ng/ml respectively). In short subjects there was a significant increase in serum IGF-I levels after the 4th GH injection (from 192.1+/-18.3 ng/ml, i.e. -1.16+/-0.16 standard deviation score (SDS) to 338.2+/-27.1 ng/ml, i.e. 0.14+/-0.17; P<0.00001). No significant differences were found between short children and controls in basal concentrations of serum INF-gamma (19+/-4 and 26+/-5 mIU/ml respectively), IL-1alpha (24.950+/-3.613 and 20.896+/-2.778 pg/ml respectively), IL-2 (3.945+/-1.209 and 4.794+/-0.562 pg/ml respectively), IL-12 (1.093+/-0.269 and 1.976+/-0.596 pg/ml respectively), and TNF-alpha (1.794+/-0.559 and 2.188+/-0.346 pg/ml respectively). Likewise, a significant increase was found in serum INF-gamma (before 19+/-4 and after four GH injections 185+/-57 mIU/ml respectively; P<0.008), IL-1beta (24.950+/-3.613 to 43.339+/-5.431 pg/ml respectively; P<0.0001), IL-2 (3.945+/-1.209 to 9.165+/-2.331 pg/ml respectively; P<0.003), IL-12 (1.093+/-0.269 to 3.724+/-0.637 pg/ml respectively; P<0.0007) and TNF-alpha (1.794+/-0.559 to 9.266+/-3.066 pg/ml respectively; P<0.01). CONCLUSIONS: Cytokine release can be affected by short-term GH administration in normal children indicating a direct influence of GH on the immune system.     

内毒素血症时肝窦内皮细胞脂多糖受体CD14蛋白表达的观察

目的观察内毒素血症时肝窦内皮细胞(LSECs)中CD14蛋白合成和CD14 基因的表达,以及CD14蛋白在内毒素介导LSECs激活中的作用. 方法经尾静脉注入脂多糖(LPS,E coli O111B4)5mg/kg,建立大鼠内毒素血症动物模型,分别于术后0(对照组)、3、6、12、24h活杀取材.用兔抗鼠CD14抗体和异硫氢酸荧光素(FITC)标记的羊抗兔IgG对LSECs进行孵育后,流式细胞仪测定LSECs的平均荧光强度(MFI)及FITC阳性细胞数;用原位杂交法测定LSEC中CD14 mRNA的表达.用原位胶原酶灌注法分离大鼠LSECs,用不同浓度LPS(0、0.01、1、10、100μg/ml)刺激LSECs.并用CD14抗体阻断LSECs的 CD14蛋白后,再用不同浓度LPS(0、0.01、1、10、100μg/ml)刺激LSECs.测定LPS介导LSECs肿瘤坏死因子(TNF)-α及白细胞介素-6(IL-6)分泌及CD14抗体对LSECs细胞因子分泌的影响. 结果内毒素血症大鼠3、6、12和24h 时LSECs的MFI明显增加;FITC阳性细胞数也明显增多,分别为54.32%、65.83%、85.61%和45.65%,与对照组的4.45%比较差异有非常显著意义(P＜0.01).原位杂交显示,内毒素血症大鼠LSECs中CD14 mRNA的表达明显增强,而对照组CD14 mRMA无阳性表达.LPS组TNF-α的含量(pg/ml)分别为54.49±6.02、84.65±10.16、206.54±23.55、349.87±39.47和365.76±40.31;CD14阻断组TNF-α的含量(pg/ml)分别为55.93±6.95、63.32±7.81、85.34±9.72、112.75±13.54、198.66±21.54;两组间比较差异有非常显著意义(P＜0.01).LPS组IL-6的含量(pg/ml)分别为103.34±12.52、187.39±20.31、243.87±27.83、289.51±30.15、298.53±31.94;CD14阻断组IL-6的含量(pg/ml)分别为104.37±11.49、125.02±13.58、164.59±19.47、183.47±20.17、221.76±26.43;两组间比较差异有非常显著意义(P＜0.01). 结论内毒素血症时LSECs能合成CD14蛋白及表达CD14基因;抗CD14抗体对LPS诱导LSECs TNF-α和IL-6的分泌有抑制作用;CD14蛋白的表达在内毒素介导LSECs激活中可能起重要作用.     

Effects of splenectomy in patients with cirrhosis undergoing hepatic resection for hepatocellular carcinoma

AIM: To study the effects of splenectomy in patients with cirrhosis undergoing hepatic resection for hepatocellular circinoma.METHODS: Twenty-six patients with HCC associated with cirrhosis were divided into hepatectomy with splenectomy group (splenectomy group, n=11) and hepatectomy without splenectomy group (non-splenectomy group, n=15). T lymphocyte subsets such as CD4, CD8, CD4/CD8, helper T (Th) lymphocyte cytokines such as interferon γ(IFN-γ),interleukin 2(IL-2), interleukin 10(IL-10) and white blood cell (WBC), platelet (PLT), total bilirubin (T-Bil) were measured and used as parameters to evaluate the effects of splenectomy.RESULTS: There was no significant difference in CD4, CD8,CD4/CD8, IL2, IFN-γ, IL10, WBC, PLT, T-Bil levels between two groups before surgery. Two months after operation,the levels of CD4 (41.2 %±4.2 % vs 34.7 %±3.8 %), CD4/CD8 (1.7±0.2 vs 1.0±0.2), IFN-γ (102.3±15.9 pg/ml vs 86.5±14.8 pg/ml), IL-2(97.2±15.6 pg/ml vS77.6±14.5 pg/ml) were increased and those of CD8 (25.6±3.9 vs32.8 %±4.1%),IL-10 (56.9±10.4 pg/ml vs 72.6±15.3 pg/ml) were decreased in splenectomy groups as compared with those in nonsplenectomy group (P<0.05). WBC and PLT counts in the splenectomy group were 8.9±1.6×109 and 310±32×109,respectively, which were significantly higher than those in non-splenectomy group (3.7±1.4×109 and 104±41×109) respectively on the 14th post-operative day. T-Bil concentration in the splenectomy group (24±7 μmol/L) was significantly lower than that in the non-splenectomy group (37±13 μmol/L) on the 7th post-operative day (P<0.05).CONCLUSION: Splenectomy combined with hepatectomy for HCC associated with cirrhosis is helpful for the recovery of T-lymphocyte subsets and the maintenance of Th:L/Th2 cytokine balance.     

Physical training modulates proinflammatory cytokines and the soluble Fas/soluble Fas ligand system in patients with chronic heart failure

OBJECTIVES: We sought to investigate the effects of physical training on circulating proinflammatory cytokines and the soluble apoptosis mediators Fas (sFas) and Fas ligand (sFasL) in patients with chronic heart failure (CHF). BACKGROUND: Recent investigations have shown an overexpression of circulating proinflammatory cytokines and soluble apoptosis mediators in patients with CHF, which may be related to their exercise intolerance and clinical deterioration. METHODS: Plasma levels of tumor necrosis factor-alpha (TNF-alpha), soluble TNF receptors I and II (sTNF-RI and sTNF-RII, respectively), interleukin-6 (IL-6), soluble IL-6 receptor (sIL-6R), sFas and sFasL were measured in 24 patients with stable CHF (New York Heart Association functional class II/III; left ventricular ejection fraction 23.2 +/- 1.3%) and in 20 normal control subjects before and after a 12-week program of physical training in a randomized, crossover design. Functional status of patients with CHF was evaluated by using a cardiorespiratory exercise test to measure peak oxygen consumption (VO2max). RESULTS: Physical training produced a significant reduction in plasma levels of TNF-alpha (7.5 +/- 1.0 pg/ml vs. 4.6 +/- 0.7 pg/ml, p < 0.001), sTNF-RI (3.3 +/- 0.2 ng/ml vs. 2.7 +/- 0.2 ng/ml, p < 0.005), sTNF-RII (2.6 +/- 0.2 ng/ml vs. 2.3 +/- 0.2 ng/ml, p = 0.06), IL-6 (8.3 +/- 1.2 pg/ml vs. 5.9 +/- 0.8 pg/ml, p < 0.005), sIL-6R (34.0 +/- 3.0 ng/ml vs. 29.2 +/- 3.0 ng/ml, p < 0.01), sFas (5.5 +/- 0.7 ng/ml vs. 4.5 +/- 0.8 ng/ml, p = 0.05) and sFasL (34.9 +/- 5.0 pg/ml vs. 25.2 +/- 4.0 pg/ml, p < 0.05), as well as a significant increase in VO2max (16.3 +/- 0.7 ml/kg per min vs. 18.7 +/- 0.8 ml/kg per min, p < 0.001). Good correlations were found between a training-induced increase in VO2max and a training-induced reduction in levels of the proinflammatory cytokine TNF-alpha (r = -0.54, p < 0.01) and the apoptosis inducer sFasL (r = -0.57, p < 0.005) in patients with CHF. In contrast, no significant difference in circulating cytokines and apoptotic markers was found with physical training in normal subjects. CONCLUSIONS: Physical training reduces plasma levels of proinflammatory cytokines and the sFas/sFasL system in patients with CHF. These immunomodulatory effects may be related to the training-induced improvement in functional status of patients with CHF.     

白细胞介素12调节小鼠Th1/Th2的抗结核分支杆菌感染研究

目的 观察、评价白细胞介素１２（ＩＬ－１２）对结核分支杆菌感染小鼠细胞因子的影响和疗效。方法 将ＢＡＬＢ／ｃ小鼠４２只,制成小鼠结核分支杆菌感染模型,随机分为对照组（ＰＢＳ）和ＩＬ－１２共两组（２１只）。给予ＰＢＳ或ＩＬ－１２治疗,检测血清γ干扰素（ＩＮＦ－γ）、ＩＬ－４、ＩＬ－１０水平（ＥＬＩＳＡ法）和器官菌数计数,观察治疗后生存率。结果 ＩＬ－１２组小鼠无一死亡、肺、肝、脾菌落数分别为（２２７     

Effects of carvedilol on plasma levels of interleukin-6 and tumor necrosis factor-alpha in nine patients with dilated cardiomyopathy

OBJECTIVES: Whether beta-blocker therapy changes the circulating levels of cytokines as congestive heart failure improves remains uncertain. METHODS: Nine patients with idiopathic dilated cardiomyopathy, who had previously received conventional treatment and were classified as New York Heart Association (NYHA) functional class II, received carvedilol by stepwise dose increase up to 20 mg daily, and the plasma interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) levels were measured. RESULTS: IL-6 was significantly reduced from 0.80 +/- 0.49 pg/ml before therapy to 0.21 +/- 0.08 pg/ml after carvedilol was increased to 20 mg daily (p < 0.05). Moreover, IL-6 level had already decreased significantly compared to the baseline when the dose of carvedilol had reached 10 mg daily (0.28 +/- 0.12 pg/ml, p < 0.05). TNF-alpha levels did not change significantly. CONCLUSIONS: These results demonstrate that IL-6 concentration is significantly decreased by beta-blocker therapy. The efficacy for heart failure may be related to the change of IL-6 concentration.     

Plasma levels of interleukin 2, 6, 10 and phenotypic characterization of circulating T lymphocytes in ischemic heart disease.

The purpose of this study was to assess lymphocyte receptors expression in patients with ischemic heart diseases, as well as to measure the plasma levels of interleukin (IL) 2, 6 and 10. T Lymphocytes are found in large numbers in human atherosclerotic plaques, indicating that immune and inflammatory mechanisms are important factors in the pathogenesis of atherosclerosis. Recent data have also implicated T lymphocytes in the pathogenetic mechanism of unstable angina and ischemic heart disease. Three groups of patients were studied: 42 with an acute ischemic syndrome (AIS), 36 with stable angina (SA) and 39 healthy controls. To characterize lymphocyte phenotype, flow cytometry was performed in whole-blood samples. IL-2, IL-6 and IL-10 were measured using the ELISA method. Double fluorescence evaluation showed an increase in CD8+/CD11b+ cells (cytotoxic T lymphocytes) and in CD11b+/CD16+CD56+ cells (NK lymphocytes) in the AIS group and in SA group as compared to the control group (P < 0.05 and P < 0.001, respectively). IL-2 was increased in the AIS and SA groups compared to the control group (AIS 4.5 +/- 0.5 pg/ml; SA 6.3 +/- 0.6 pg/ml; controls 2.4 +/- 0.8 pg/ml, P < 0.05), whereas IL-6 was higher in the AIS group than in the other two groups (AIS 10.8 +/- 1.8 pg/ml; SA 1.8 +/- 0.8 pg/ml; controls 1.2 +/- 0.6 pg/ml, P < 0.0001). These data show that patients with ischemic heart disease have an increase in circulating cytotoxic T lymphocytes and in IL-2 plasma levels, irrespective of their clinical presentation, compared to normal control subjects, whereas IL-6 is elevated only in patients with AIS.     

冠心病患者分泌型磷脂酶A2的变化及其与白细胞介素8、溶血磷脂酸的关系

目的了解冠心病患者分泌型磷脂酶 A2(sPLA2)的变化,并通过测定 IL-8、LPA 等相关细胞因子,对 sPLA2的作用机制进行初步探讨。方法冠状动脉造影确诊的262例患者,其中急性冠脉综合征(ACS)110例,稳定性冠心病(SCHD)63例,正常患者89例,分别测定 sPLA2、白细胞介素8(IL-8)、高敏 C 反应蛋白(hs-CRP)和溶血磷脂酸(LPA)水平,并进行对比分析。结果冠心病患者sPLA2[(68±17)U/ml]、IL-8[(182±80)pg/ml]以及 LPA[(2.85±0.36)μmol/L]均明显高于对照组[sPLA2:(55±12)U/ml,IL-8:(119±33)pg/ml,LPA:(2.34±0.36)μmol/L,均 P<0.01],其中ACS 组的 sPLA2[(71±18)U/ml]、IL-8[(195±78)pg/ml]也明显高于 SCHD 组[sPLA2:(63±12)U/ml],IL-8:[(159±79)pg/ml,均 P<0.01;sPLA2与 IL-8(r=0.203,P=0.007)、LPA(r=0.658,P<0.01)、hs-CRP(r=0.231,P=0.005)均呈正相关;sPLA2≥63.75 U/ml 时,患冠心病的相对危险度为6.248(P<0.01)。结论冠心病患者 sPLA2明显升高,它可能与 IL-8共同参与冠心病的炎症发展过程,并与下游相关产物 LPA 进一步加速其进展。提示 sPLA2升高是冠心病的独立危险因素之一。     

Effects of growth hormone on circulating cytokine network, and left ventricular contractile performance and geometry in patients with idiopathic dilated cardiomyopathy.

BACKGROUND: Recent experimental and clinical data indicate that abnormal central and peripheral immune reactions contribute to the progression of chronic heart failure, and that immunomodulation may be an important therapeutic approach in this syndrome.Aims We sought to study the effects of growth hormone (GH) administration on circulating pro-inflammatory/anti-inflammatory cytokine balance, and to investigate whether these GH-induced immunomodulatory effects are associated with the improvement of left ventricular (LV) contractile performance in idiopathic dilated cardiomyopathy (DCM) patients. METHODS: Plasma pro-inflammatory cytokines tumour necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), granulocyte-macrophage colony-stimulating factor (GM-CSF) and its soluble receptor (sGM-CSFR), chemotactic cytokine macrophage chemoattractant protein-1 (MCP-1), soluble adhesion molecules intercellular adhesion molecule-1 (sICAM-1) and vascular cell adhesion molecule-1 (sVCAM-1), and, finally, anti-inflammatory cytokines interleukin-10 (IL-10) and transforming growth factor-beta2 (TGF-beta2) were measured (ELISA method) in 12 patients with DCM (NYHA class III; LV ejection fraction: 23.6+/-1.7%) before and after a 3-month subcutaneous administration of GH 4IU every other day (randomized crossover design). Peak oxygen uptake (VO2 max), LV dimensions, LV mass index, end-systolic wall stress (ESWS), mean velocity of circumferential fibre shortening (Vcfc), and contractile reserve (change of ratio Vcfc/ESWS after dobutamine administration) were also determined at the same period. RESULTS: Treatment with GH produced a significant reduction in plasma TNF-alpha (7.8+/-1.1 vs 5.5+/-0.9pg/ml, P=0.013), IL-6 (5.7+/-0.5 vs 4.7+/-0.4pg/ml, P=0.043), GM-CSF (27.3+/-1.7 vs 23.3+/-1.8pg/ml, P=0.042), sGM-CSFR (4.0+/-0.4 vs 3.2+/-0.4ng/ml, P=0.039), MCP-1 (199+/-5 vs 184+/-6pg/ml, P=0.048), sICAM-1 (324+/-34 vs 274+/-27ng/ml, P=0.008) and sVCAM-1 (1238+/-89 vs 1043+/-77ng/ml, P=0.002) in DCM patients. A significant increase in ratios IL-10/TNF-alpha (1.9+/-0.3 vs 3.5+/-0.9, P=0.049), IL-10/IL-6(2.6+/-0.6 vs 3.2+/-0.5, P=0.044) and TGF-beta2/TNF-alpha (3.1+/-0.6 vs 4.4+/-0.6, P=0.05) was alsofound with GH therapy. A significant reduction in ESWS (841+/-62 vs 634+/-48gr/cm(2), P=0.0026) and LV end-systolic volume index (LVESVI, 128+/-12 vs 102+/-12ml, P=0.035) as well as a significant increase in posterior wall thickness (PWTH, 9.2+/-0.5 vs 10.3+/-0.6mm, P=0.034), contractile reserve (0.00029+/-0.0001 vs 0.00054+/-0.0001circ*cm(2)/gr*s, P=0.00028) and VO2max (15.3+/-0.7 vs 17.1+/-0.9ml/kg/min, P=0.002) were observed after GH administration. Good correlations were found between GH-induced increase in contractile reserve and the increases in VO2max (r=0.63, P=0.028), IL-10/TNF-alpha (r=0.69, P=0.011) and TGF-beta2/TNF-alpha (r=0.58, P=0.046) ratios, as well as the reduction in plasma TNF-alpha levels (r=-0.86, P=0.0004). CONCLUSIONS: GH administration modulates beneficially circulating cytokine network and soluble adhesion molecules in patients with DCM, whilst enhancing contractile reserve and diminishing LV volumes. These GH-induced anti-inflammatory effects may be associated with the improvement in LV contractile performance and exercise capacity as well as with the reverse of LV remodelling of patients with DCM.     

Effects of hyaluronate on CD44 expression of infiltrating cells in exudate of rat air pouch, induced by sensitization with lipopolysaccharide.

OBJECTIVE: To investigate the effects of hyaluronate (HA) on CD44 expression of infiltrating cells in vivo. METHODS: Intra-air pouch injection of 10 microg lipopolysaccharide (LPS) with 0.4, 4.0, or 40 mg HA, 40 mg carboxymethylcellulose (CMC), or saline was performed on rats immunized with LPS. The percentage of CD44+ cells in the exudate of the air pouch was measured by flow cytometry, and the concentrations of interleukin 1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) in the air pouch exudate were measured by ELISA. IL-1beta and TNF-alpha in the air pouch lining layer were stained by immunohistochemistry. RESULTS: The percentage of CD44+ cells in air pouch exudate was greater in the presence of HA, with a dose dependent increase (0.4 mg, 9.4+/-2.6%, n = 4; 4.0 mg, 13.8+/-2.9%, n = 4; 40 mg, 24.9+/-6.3%, n = 3; p < 0.05), while it was 4.9+/-1.2% (n = 4) in the presence of 40 mg CMC. The concentration of IL-1beta was lower in the presence of 40 mg HA (251.0+/-61.4 pg/ml, n = 4) or 40 mg CMC (168.2+/-43.5 pg/ml, n = 4; p < 0.05) than in saline (403.0+/-60.5 pg/ml, n = 4). The concentration of TNF-alpha was lower in the presence of 40 mg HA (14.0+/-6.7 pg/ml, n = 4) or 40 mg CMC (7.04+/-7.0 pg/ml, n = 4) than in saline (38.2+/-12.2 pg/ml, n = 4). Extensively stained lining cells in superficial layer of the air pouch with IL-1beta and TNF-alpha were observed in rats inoculated with 0.4 mg HA. CONCLUSION: These findings suggest that HA might affect the inflammatory process through modifying CD44 expression on infiltrating cells in air pouch exudate.     

Hepatitis C Virus and related changes in immunological parameters in non Hodgkin's lymphoma patients

Viral hepatitis is a common and important problem in immunocompromised cancer patients. The present study was conducted to investigate changes in some cellular and humoral immunological parameters as a consequence of HCV infection in non Hodgkin's lymphoma patients (NHL). The study included 40 NHL patients: 20 anti-HCV antibody positive (Gr. I ), and 20 anti-HCV antibody negative (Gr.II ). In addition, forty non-cancer controls (NCCs) were included: 20 of them were anti-HCV antibody positive (Gr. III) and 20 anti-HCV antibody negative (Gr. IV). The studied immunological parameters included serum levels of interleukin-1 (IL-1), interleukin-2 (IL-2), interleukin-6 (IL-6), and soluble tumor necrosis factor receptors (s-TNFr) measured by ELISA, as well as assessment of T and B lymphocyte subsets by PAP immunostaining method. Mean IL-1 level (pg/ml) was significantly higher in Gr. 1 (14 +/- 6) and Gr. III (20 +/- 12) as compared to those in Gr. II (7 +/- 5) and Gr. IV (9 +/- 6). Mean IL-2 level (pg/ml) was also significantly higher in Gr. I (132 +/- 101) and Gr. III (135 +/- 59) compared to those in Gr. II (36 +/- 29) and Gr. IV (31 +/- 48). On the other hand, level of IL-6 showed no significant difference between groups. The mean level of sTNF-r, (ng/ml) was only significantly higher in Gr. I (2.9 +/- 1.7) when compared to that in Gr. IV (1.9 +/- 2.2). In group IV, the average percentage of CD3 (70 +/- 4%) and CD4 (44 +/- 5%) were significantly higher than in those of Gr. I (CD3 = 51 +/- 11%, CD4 = 30 +/- 12%), Gr. II (CD3 = 52 +/- 7%, CD4 = 30 +/- 8%), and Gr. III (CD3 = 52 +/- 9%, CD4 = 26 +/- 8%). From all the above immunological and virological features two main tips could be inferred: (1) HCV leads a mild course of infection in NCCs evidenced by normal ALT level in all but 20% of subjects, normal IL-6, sTNF-r, lower counts of CD4+ T cells and hence a mild hepatocellular injury, and (2) In the immunocompromised NHL patients the virus leads potentially more aggressive course as evidenced by higher viremia, as well as significant elevation in sTNF-r, and CD8+ depression.     

弓形虫感染大鼠外周血T淋巴细胞亚群及IFN-γ、TNF-α、IL-4动态变化

目的 观察弓形虫感染大鼠外周血T淋巴细胞亚群及干扰素-γ（IFN-γ）、肿瘤坏死因子-α（TNF-α）、白细胞介素-4（IL-4）动态变化。 方法 将54只清洁级Sprague-Dawley（SD）成年大鼠随机分成9组,其中8组每鼠腹腔注射经纤维素-11（CF-11）纯化的活弓形虫速殖子悬液2 ml（2×10⁵/L）。分别于弓形虫感染后1、3、7、14、28、35、42、60 d时取大鼠外周全血,流式细胞术检测T细胞亚群变化,ELISA检测外周血IFN-γ、TNF-α、IL-4动态变化。对照组腹腔注射2 ml生理盐水。 结果 T淋巴细胞亚群CD8+水平在感染弓形虫后3 d（14.22%）、7 d（14.93%）、14 d（16.08%）均显著低于正常水平（23.08%）（P＜0.05）,至感染后28 d恢复正常水平,而CD4+水平在整个感染过程中无明显变化。IFN-γ和IL-4水平在感染后第7 d分别升高至6.73 pg/ml和6.91 pg/ml（P＜0.05）,至感染60 d仍高于正常水平;TNF-α至感染后28 d（14.37 pg/ml）及60 d均高于正常水平（10.81 pg/ml）（P＜0.05）。 结论 弓形虫感染后大鼠外周血CD8+水平一度降低,28 d后恢复正常,CD4+无明显变化,IFN-γ、IL-4和TNF-α均高于正常水平,但出现时间有差异。     

The increase of parathyroid hormone-related peptide and cytokine levels in synovial fluid of elderly rheumatoid arthritis and osteoarthritis

We simultaneously measured the concentrations of parathyroid hormone related peptide (PTHrP) and cytokines in synovial fluid (SF) to clarify the relationship between PTHrP and cytokine network in the SF of elderly patients with arthritis. SF was collected from knee joints of five RA patients aged 66+/-11 years old and nine osteoarthritis (OA) patients aged 80+/-9 years old. PTHrP in SF was measured by enzyme-linked immunosorbent assay (ELISA), whereas tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-6 (IL-6) and interleukin-8 (IL-8) in SF were all measured by ELISA. The PTHrP levels in the SF of RA patients (2.56+/-0.89 pmol/l) were significantly (p<0.05) higher than those of OA patients (1.66+/-0.17 pmol/l). TNF-alpha, IL-1beta, IL-2 and IL-6 concentrations in SF of RA were also significantly higher than those in SF of OA (TNF-alpha 22.5+/-14.8 vs 4.8+/-3.0 pg/ml, p<0.01; IL-1beta 11.8+/-11.4 vs 1.4+/-1.3, p<0.05; IL-2 59.9+/-46.6 vs 12.5+/-8.0 pg/ml, p<0.05; IL-6 18424+/-8901 vs 3547+/-2948 pg/ml, p<0.01). The concentrations of IL-4 and IL-8 in SF of RA were similar to those of OA. Immunohistochemical studies revealed the presence of immunoreactive PTHrP in synovial fibroblasts from RA and OA. Among cytokines, only IL-6 was positively correlated with PTHrP levels in SF (r=0.685, p<0.01). In the culture of synovial cells from RA and OA, PTHrP was produced in RA more than OA after phorbol 12-mysistate 13-acetate (TPA) stimulation. These results indicate that PTHrP and cytokines, especially IL-6, might be involved in the inflammatory processes of elderly RA and OA. This is the first study in which PTHrP and cytokine levels were simultaneously examined in synovial fluid of elderly RA and OA.     

Differentiation of dendritic cells in monocyte cultures isolated from patients with unstable angina

OBJECTIVE: Dendritic cells (DC) stimulate T-cell proliferation and activation in the course of adaptive immunity. Its role in unstable coronary plaque in humans is unknown. So we investigated the functional role of DC in patients with unstable angina pectoris (US) using human monocyte-derived DC. METHODS: Twenty milliliters of blood was drawn from the femoral artery of 20 patients, who underwent coronary angiography. Ten patients with a diagnosis of US and 10 patients with normal CAG were included in the observation and control groups, respectively. The mononuclear cells were separated from the peripheral blood and cultured in RPMI1640 with supplement of rh GM-CSF and rh IL-4 to induce DC. The shape and ultrastructure of DC was analyzed with electronic microscopy. The phenotype of DC was analyzed with FACS and the alloantigen presenting capacity of DC was evaluated by mixed lymphocyte reaction (MLR). The levels of cytokines in allogenic DC/T cell cultures were assayed by ELISA. RESULTS: The expression rate of CD86 in patients with US was 30.8+/-3.3%, which was obviously higher than that of normal DC (19.4+/-3.0%), P<0.001. The capacity of proliferation of patients DC to induce allogenic T cells (OD 1.82+/-1.29), which was obviously higher than that of the normal DC (OD 0.81+/-0.41), P<0.005. TNF-alpha (40.05+/-7.15 pg/ml), IL-1beta (19.01+/-1.39 pg/ml) and IL-6 (40.80+/-16.04 pg/ml), produced during MLR were higher in patients with US than that of normal patients 7.85+/-1.10, 12.18+/-1.93 and 19.55+/-0.7, respectively, P<0.05. IL-10 (10.94+/-0.56 pg/ml) produced during MLR was lower in patients with US than that of normal patients (16.63+/-3.40 pg/ml), P<0.05. CONCLUSION: Our results suggest that the function of DC in patients with US is increased and these activated effects of DC may play a primary role in the immune process of plaque rupture.     

High serum tumor necrosis factor-alpha levels are associated with lack of response to infliximab in fistulizing Crohn's disease

OBJECTIVES: Infliximab, a chimeric monoclonal antibody directed against tumor necrosis factor-alpha (anti-TNF-alpha), has been effective in the treatment of patients with active Crohn's disease and with fistulas. We investigated the effect of infliximab on circulating cytokines and acute phase proteins in patients with fistulas to determine the clinical response to anti-TNF-alpha. METHODS: A total of 36 patients with fistulizing Crohn's disease were selected for study. Serum from patients was drawn before the infusion on day 0 and at wk 2, 4, 6, 8, and 10 after completion of treatment. Circulating concentrations of TNF-alpha, interleukin-1beta (IL-1beta), and IL-6 were measured by ELISA. The functional activity of circulating TNF-alpha was assessed by the WEHI 164 TNF-alpha bioassay. Acute phase proteins were also determined. RESULTS: Elevated TNF-alpha, IL-1beta, IL-6, and acute phase proteins were observed in patients with Crohn's disease. Of the patients with fistulas, 22 (61.1%) responded to treatment. Before receiving infliximab, higher levels of serum TNF-alpha were found in patients who did not respond to infliximab compared with those who did (median interquartile range 26, 0-245 pg/ml; n = 14 vs 0, 0-22 pg/ml, n = 22). Patients showed no change in circulating levels of TNF-alpha during the course of the study. CONCLUSIONS: This treatment produces a clinical improvement in about two-thirds of CD patients with fistulas. The circulating levels of TNF-alpha are associated with the response to infliximab and could help to identify patients who would benefit from anti-TNF-alpha treatment.     

持续吸入变应原引起支气管哮喘小鼠免疫耐受

目的探讨吸入变应原引起支气管哮喘(简称哮喘)过敏性气道炎症免疫耐受形成的机制。方法BALB/c小鼠60只,按随机数字表法分为实验组(50只)和空白对照组(10只),实验组小鼠先给予腹腔注射卵清白蛋白(OVA)1mg,每周1次,共3周。雾化吸入OVA每天1h(含OVA80μg),连续10d。依据吸入OVA时间分为A、B、C、D、E5组,每组10只。A组雾化吸入10d后处死。B、D组继续每天1次,每次1h,每周5次,分别吸入OVA4周及8周,然后每天1h,连续10d吸入OVA后处死。C组停止吸入OVA4周后再次吸入OVA,每天1h,连续10d后处死。E组每天1次,每次1h,每周5次,吸入OVA4周,停止雾化吸入OVA4周,然后每天1h,连续10d吸入OVA后处死。测定各组小鼠支气管肺泡灌洗液(BALF)中细胞总数,嗜酸粒细胞、淋巴细胞、CD4+、CD8+、CD4+IL-10+分类及BALF中白细胞介素4(IL-4)、γ干扰素(IFN-γ)、IL-10的含量。测定血清中IL-4、IFN-γ、IL-10、OVA、IgE、IgG1、IgG2a水平,并对各组小鼠肺组织病理学进行分析。结果空白对照组BALF中嗜酸粒细胞、B淋巴细胞、CD4+IL-10+细胞分别为0.010±0.000、2.1±1.9、4.9±1.5,A组分别为0.480±0.110、5.1±2.6、5.1±2.3,B组分别为0.120±0.020、8.9±3.6、10.4±3.6,C组分别为0.560±0.050、4.7±1.7、6.3±3.1,D组分别为0.070±0.030、10.1±2.9、12.7±4.5,E组分别为0.680±0.030、5.6±3.2、6.1±3.4,各组间比较差异有统计学意义(F值分别为36.46、31.89、167.89,P均<0.01)。B、D组BALF中CD4+IL-10+细胞数与A组比较差异有统计学意义(q=5.8、6.4,P均<0.05);空白对照组BALF中IL-4、IL-10水平分别为(21±3)pg/ml、(44±12)pg/ml,A组分别为(128±23)pg/ml、(68±18)pg/ml,B组分别为(54±12)pg/ml、(127±27)pg/ml,C组分别为(133±21)pg/ml、(78±17)pg/ml,D组分别为(8±18)pg/ml、(135±34)pg/ml,E组分别为(143±26)pg/ml、(76±15)pg/ml,组间比较差异有统计学意义(F分别为37.20、143.78,P均<0.01)。B、D两组BALF中IL-10水平与A组比较差异有统计学意义(q分别为7.8、9.6,P均<0.05)。结论持续吸入变应原可使小鼠气道炎症减轻,产生免疫耐受,调节T淋巴细胞产生的IL-10参与了耐受形成。     

TNF-alpha levels in children with growth hormone deficiency and the effect of long-term growth hormone replacement therapy.

OBJECTIVE: Growth hormone (GH) has been suggested to modulate the release of some cytokines including TNF-alpha. To investigate TNF-alpha levels in children with GH deficiency (GHD), to evaluate alteration in TNF-alpha levels during recombinant human GH (rhGH) treatment, and to analyze possible correlations between TNF-alpha and GH, IGF-1 and IGFBP-3. DESIGN: Twenty-four children, aged 12.60+/-2.27 years, with isolated GHD and given rhGH therapy, as subcutaneous ingestion of 0.03-0.04mg/kg once-daily dose, were evaluated. Eleven had complete and 13 had partial GHD. Thirty-three healthy children were studied as controls. Age and sex distribution, body mass indexes of two groups were similar. In children with GHD, blood samples were drawn before (TNF-alpha0), and at 6 (TNF-alpha6) and 12 (TNF-alpha12) months of the treatment with rhGH. TNF-alpha was determined using a human TNF-alpha ELISA assay (Biosource International). RESULTS: TNF-alpha0 levels were significantly higher in children with GHD than in controls (41.79+/-25.04 and 8.63+/-4.48pg/ml, respectively, p<0.001) and decreased significantly during rhGH treatment (TNF-alpha0=41.79+/-25.04, TNF-alpha6=13.67+/-9.95, TNF-alpha12=10.86+/-6.61pg/ml, p<0.05). There was no correlation between TNF-alpha levels and BMI, IGF-1/logIGF1, IGFBP-3 levels and growth velocity of the patients with GHD. Although no correlation between TNF-alpha and peak GH levels after stimulation was present; a moderate reverse correlation between TNF-alpha and basal serum concentrations of GH (r=-0.512, p=0.046) was demonstrated. CONCLUSIONS: TNF-alpha levels are significantly higher in children with GHD than the controls, and long-term therapy with rhGH effectively reduces its level. Our data suggest that GH plays an inhibitory role on TNF-alpha release in humans. However, due to inconsistent results up to now, further prospective, controlled and long term studies are needed to elucidate the issue.