甘氨酸对内毒素性肝损害保护作用的机制

目的:探讨甘氨酸(Gly)对内毒素(LPS)性肝损害的保护机制．方法:BALB／c小鼠随机分为三组,LPS组(n= 50)经腹腔注射10 mg／kg的LPS,Gly组(n=50) 在注射相同剂量LPS前3 d开始喂饲含50 g／L 的Gly的饲料,正常生理盐水对照组(n=50), 经腹腔注射等体积的生理盐水,光镜观察组织病理学改变,免疫组织化学法检测TLR4表达水平:ELISA法检测血浆TNF-α,IL-10浓度及 RT-PCR检测肝组织中TNF-α,IL-10及TLR4的 mRNA表达水平．结果:Gly能明显提高小鼠存活率,肝脏病理损害程度减轻:Gly组TNF-α水平显著低于LPS 组,差异有统计学意义(708．83±51．29 ng／L vs 1852．8±126．64 ng／L,F=786．21,P<0．05);Gly 组IL-10N加且高峰前移,与LPS组比较差异有统计学意义(418．64±38．86 ng／L vs 211．15 ±26．44 ng／L,P<0．05);Gly组肝组织中TNF-α及TLR4表达也明显减弱,IL-10表达明显增强, 与LPS组比较差异均有统计学意义(分别为 TNF-α A值:1．59±0．14 vs 0．91±0．11;TLR4 A值:0．97±0．12 vs 0．53±0．11;IL-10A值:0．62 ±0．08 vs 1．06±0．15;P均<0．05)．结论:Gly能明显减轻LPS所致的肝损害,其机制可能与其下调肝细胞的TLR4表达,同时上调IL-10的水平有关．     

羟基红花黄色素A缓解脂多糖诱导内皮细胞炎症因子表达升高的研究

目的:研究羟基红花黄色素A(HSYA)抑制脂多糖(LPS)诱导的脐静脉血管内皮细胞(Eahy926细胞)炎症因子表达升高的作用。方法:采用RT-qPCR法测定Toll样受体4(TLR4)、白介素6(IL-6)、白介素1β(IL-1β)和肿瘤坏死因子α(TNFα)mRNA表达水平及ELISA法测定IL-6、IL-1β和TNFα蛋白表达水平。结果:HSYA浓度为5×10-6mol/L、10×10-6mol/L和20×10-6mol/L时可抑制LPS(终浓度为1μg/mL)诱导的Eahy926细胞IL-6、IL-1β和TNFαmRNA(模型组vs.正常组P值均<0.01;HSYA干预中高剂量组vs.模型组P值均<0.01;HSYA干预低剂量组vs.模型组,P值均<0.05)和TNFα蛋白(模型组vs.正常组,P值均<0.01;HSYA干预IL-1β表达的中高剂量组vs.模型组,P值<0.05;HSYA干预TNFα表达的中剂量组vs.模型组,P值<0.05;高剂量组vs.模型组,P值<0.01。HSYA干预IL-6表达的低剂量组vs.模型组,P值<0.05;HSYA干预中高剂量组vs.模型组,P值<0.01)表达的升高,并随HSYA剂量升高可见药效增强。结论:HSYA对LPS诱导的Eahy926细胞TLR4、IL-6、IL-1β、TNFα表达升高有抑制作用。     

CD14和Toll样受体4的表达在内毒素致肝炎重型化中的意义

目的 探讨CD14和Toll样受体4(TLR4)在内毒素(LPS)诱导机体炎性因子分泌过程中的作用,阐明乙型肝炎重型化的机制. 方法 采用流式细胞学方法和逆转录聚合酶链反应等技术,检测30例慢性重型乙型肝炎患者,30例慢性乙型肝炎患者和20名健康者(对照组)的外周血单核细胞的mCD14水平及CD14 mRNA,TLR4 mRNA表达水平,同时应用动态浊度法测定患者血浆LPS水平,应用酶联免疫吸附法检测患者血清肿瘤坏死因子(TNF)α、白细胞介素(IL)-1 β,IL-6的含量.数据的统计分析采用SPSS11.5统计软件完成,分别采用单因素方差分析、非参数检验、Nemenyi法和直线相关分析.结果 外周血单个核细胞mCD14表达水平:慢性重型乙型肝炎组为74.2%±12.3%,慢性乙型肝炎组为63.6%±11.8%,对照组为60.3%±7.20/;CD14mRNA和TLR4 mRNA的相对表达:慢性重型乙型肝炎组分别为2.92±0.67和1.86±0.45,慢性乙型肝炎组分别为1.34±0.51和0.93±0.18,对照组分别为0.92±0.58和0.73±0.16,慢性重型乙型肝炎组均明显高于慢性乙型肝炎组和对照组,F值分别为11.473、85.037和102.328,P值均<0.01,差异有统计学意义.血浆LPS、IL-l,IL-6水平:慢性重型乙型肝炎组分别为(1.87±1.61)Eu/ml,(0.96±0.16)pg/ml和(68.34±48.30)pg/ml,慢性乙型肝炎组分别为(0.11±0.11)Eu/ml、(0.19±0.02)pg/ml,(19.28±4.65)pg/ml,对照组分别为(0.03±0.03)Eu/ml、(0.15±0.01)pg/ml、(12.01±3.88)pg/ml,慢性重型乙型肝炎组均明显高于慢性乙型肝炎组和对照组,χ2值分别为32.065、83.472、36.236,P值均<0.01.外周血TNFα表达水平:慢性重型乙型肝炎组为(19.78±9.25)pg/ml、慢性乙型肝炎组分别为(7.26±6.52)pg/ml、对照组分别为(4.15±4.06)pg/ml、F=35.092,P<0.01.相关分析显示,在慢性重型乙型肝炎组LPS水平与mCD14、CD14:mRNA,TLR4 mRNA表达水平呈明显的相关性,n=0.865、r2=0.415、r3=0.524,P值均<0.05.结论 LPS可能通过以CD14和TLR4为主的LPS受体激活及其信号传导,刺激炎性因子分泌的途径,在肝炎重型化过程中发挥重要作用.     

白介素-10抑制肝星状细胞细胞间黏附分子-1的表达

目的探讨白介素-10(IL-10)对肝星状细胞(HSC)细胞间黏附分子-1(ICAM-1)表达的影响.方法采用肝脏离体胶原酶灌注消化及密度梯度离心的方法来分离培养HSC,传代后的HSC随机分为4组对照组(A组)、肿瘤坏死因子α(TNFα)100U/ml组(B组)、TNFα100U/ml+IL-102ng/ml组(C组)及TNFα100U/ml+IL-1020ng/ml组(D组).加药后24h,采用细胞酶联免疫吸附分析(ELISA)和半定量逆转录聚合酶链反应(RT-PCR)等方法,检测HSCICAM-1蛋白及mRNA表达.结果B组的ICAM-1蛋白和mRNA表达量分别为1.400±0.077和1.301±0.095,明显高于A组的0.559-0.071(P＜0001)和0.666±0.023(P＜0.001);C组和D组的ICAM-1蛋白表达量分别为1.017±0066和0.919±0.039,mRNA表达量分别为1.116±0.017和0.979±0.067,均低于B组的ICAM-1蛋白和mRNA表达(P均＜005).结论IL-10通过抑制HSCICAM-1表达,在抑制肝脏炎症、肝纤维化中发挥作用.     

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.     

Il-6及TNF-α表达与粥样斑块稳定性及冠心病的相关性分析

目的：分析IL - 6和TN F - α的表达与粥样斑块稳定性及冠心病的相关性。方法：收集我科收治的临床诊断为冠心病的患者72例,另外收集健康体检者30例,将其分为稳定性板斑块组、不稳定性斑块组及正常对照组,检测三组患者白细胞介素6及肿瘤坏死因子α的表达情况及观察冠状动脉狭窄程度的Gensini积分与IL-6及TNF-α的相关性。结果：稳定性斑块组血清IL-6为28.43±13.16pg/ml,不稳定性斑块组为38.32±18.74pg/ml,远高于对照组血清IL-6的值3.75±2.42pg/ml,稳定性斑块组血清TNF - α为19.09±7.70pg/ml,不稳定性斑块组为35.26±14.72pg/ml,远高于对照组血清TNF - α的值3.53±1.99pg/ml,差异均具有统计学意义(P<0.05),血清IL - 6值与Gensini积分成正相关(r=0.692,P=0.000),血清TN F - α值与Gensini积分成正相关(r=0.599,P=0.000)。结论：IL-6及TNF-α与粥样斑块稳定性及冠心病之间具有相关性,且IL-6及TNF-α的血清值越高,冠状动脉血管损害越严重。     

实验性酒精性肝病时脂多糖结合蛋白和脂多糖受体CD14的表达

目的观察大鼠酒精性肝病时脂多糖结合蛋白(lipopolysaccharide binding protein,LBP)和脂多糖受体CD14的表达及其在酒精性肝损害中的作用.方法随机将Wistar大鼠分为乙醇喂养组和葡萄糖喂养对照组,分剐在饮水中加入乙醇(剂量5-12 g@kg-1@d-1)和相同量的葡萄糖.两组大鼠分别于4周和8周测定其血浆中内毒揪素(LPS)浓度及血清中ALT变化,同时用RT-PCR测定肝组织中LBP和CD14 mRNA的表达,并在光镜和电镜下观察肝脏的形态学改变.结果乙醇喂养组4周和8周时大鼠血浆LPS浓度分别为(129±21)pg/ml和(187±35)Pg/m1,明显高于对照组的(48±9)pg/ml和(53±11)pg/ml(f值分别为11.2和11.6,P＜0.05);乙醇组大鼠血清ALT浓度为(112±15)U/L和(147±22)U/L,也明显高于对照组的(31±12)U/L和(33±9)U/L(t值分别为5.9和20.6,P＜0.05).乙醇组大鼠肝组织中LBP和CD14 mRNA的表达水平明显高于对照组(P＜0.05),其肝组织发生显著的病理变化,主要表现为脂肪变性、炎性细胞浸润及细胞坏死.对照组肝组织中LBP和CD14mRNA无明显表达,其病理变化也不明显.结论乙醇能诱导大鼠血中LPS浓度升高和肝缝织中LBP与CD14 mRNA的表达显著增强,增高的LBP和CD14 mRNA能增加肝脏对LPS的敏感性,可能造成肝脏损害.     

大黄甘草汤对急性坏死性胰腺炎大鼠并发的肺损伤的影响

目的 观察大黄甘草汤对急性坏死性胰腺炎(ANP)大鼠并发肺损伤的治疗效果,探讨其作用机制.方法 90只Wistar大鼠按完全随机法分为假手术组、ANP组和大黄甘草汤治疗(治疗)组,每组30只.采用逆行胰胆管注射4%牛磺胆酸钠方法制备ANP模型.治疗组在制模后给予大黄甘草汤(0.25 g/ml)0.6 ml/100 g体重灌胃,1次/12 h;假手术组与ANP组予等量生理盐水灌胃.术后6、12、24 h分批处死大鼠,取胰腺及肺组织行病理学检查并评分;测血清及肺组织IL-6、IL-10、TNF-α水平;免疫组化法检测肺组织Toll样受体4(TLR4)的表达量.结果 制模后12 h,假手术组、ANP组和治疗组的血IL-6水平分别为(14.4±4.0)pg/ml、(171.4±41.3)pg/ml、(156.9±34.7)pg/ml,IL-10水平为(13.7±4.5)pg/ml、(120.5±23.7)pg/ml、(148.3±44.4)pg/ml,TNF-α水平为(22.4±4.7)pg/ml、(261.3±51.4)pg/ml、(235.3±45.9)pg/ml;肺组织IL-6水平为(257.3±55.9)pg/g、(2578.3±403.0)pg/g、(2370.0±491.0)pg/g,IL-10水平为(80.8±20.8)Pg/g、(642.0±107.3)pg/g、(695.3±151.7)Pg/g,TNF-α水平为(207.6±98.6)pg/g、(1769.1±635.6)Pg/g、(1401.1±450.5)pg/g;胰腺病理评分为0、7.0±1.3、6.3±1.0;肺脏病理评分为0、6.3±1.4、5.6±1.0;肺组织TLR4表达量为0.09 ±0.03、0.59±0.09、0.52±0.08.ANP组和治疗组的上述指标均显著高于假手术组(P值均＜0.01);除IL-10外,治疗组的指标又显著低于ANP组(P值均＜0.05).肺组织病理学损伤与胰腺组织损伤呈正相关(r=0.807,P＜0.01),与TLR4表达水平亦呈正相关(r=0.519,P＜0.01).结论 大黄甘草汤可快速改善ANP并发的肺损伤,其机制可能与抑制TLR4的表达、降低IL-6和TNF-α、升高IL-10水平有关.     

大鼠非酒精性脂肪性肝炎形成过程中血清内毒素含量的变化

目的 探讨内毒素在非酒精性脂肪性肝炎发病中的作用。方法 通过高脂饮食建立大鼠非酒精性脂肪性肝炎(NASH)模型,并在实验第4、8、12、16、24周分批处死,同期设正常饮食组作为对照。腹主动脉采血,测定血清内毒素,肿瘤坏死因子(TNFα)和白细胞介素-1(IL-1)β水平,肝组织切片行溶菌酶、CD14免疫组织化学染色。 结果 成功建立大鼠NASH伴肝纤维化模型,NASH大鼠外周血内毒素水甲仅在24周脂肪性肝炎肝纤维化阶段明显升高为(0.23±O.06)Eu/L,对照组为(0.15±0.03)Eu/L(t>2.179,P<0.05),而4周起肝组织CD14表达就上调,溶菌酶阳性的库普弗细胞被激活,并随着实验的进展更加明显。血清TNFα水平从8周起明显增高为(26.39±24.21)pg/ml,对照组为(9.82±9.29)pg/ml(t>2.145,P<0.05),IL-1β从16周起升高为(23.76±21.81)pg/ml,对照组为(6.25±2.98)pg/ml(t>2.145,P<0.05)。 结论 NASH时存在内毒素性肝损伤。内毒素激活肝脏库普弗细胞以及促使TNFα等细胞因子释放可能是NASH的发病机制之一。     

二草清肝汤对内毒素性肝损害防护作用的实验研究

目的：探讨中药二草清肝汤对内毒素/脂多糖（LPS）性肝损害的防护机制。方法：采用LPS腹腔注射（10mg/kg）制备小鼠内毒素血症肝损害模型,BALB/c小鼠200只随机分为正常对照组、LPS组、二草清肝汤小剂量组和二草清肝汤大剂量组;光镜观察肝组织病理学改变,全自动生化分析仪检测血浆丙氨酸氨基转移酶（ALT）水平,酶联免疫吸附法检测血浆白细胞介素-12（IL-12）、肿瘤坏死因子（TNF）-α浓度,免疫组织化学法检测Toll样受体4（TLR4）表达水平,逆转录聚合酶链反应（RT-PCR）检测肝组织TLR4 mRNA表达水平。结果：二草清肝汤能明显提高小鼠存活率,肝组织病理损害程度减轻;二草清肝汤小剂量组及大剂量组小鼠的IL-12、TNF-α水平显著低于LPS组,差异有显著性意义（均P（0.05）;二草清肝汤小剂量组和大剂量组小鼠肝组织TLR4 mRNA水平也明显低于LPS组,差异均有显著性意义（均P（0.05）,但二草清肝汤大、小剂量组间差异无显著性意义（P〉0.05）。结论：二草清肝汤能明显减轻LPS所致的肝损害,其机制可能与其下调肝脏各种细胞的TLR4表达,同时下调IL-12、TNF-α的水平有关。     

Toll样受体参与小鼠肝脏缺血再灌注损伤

目的探讨Toll样受体是否参与小鼠肝脏缺血再灌注损伤及其机制. 方法用Toll样受体缺损小鼠(C3H/Hej,Hej组)和野生型(C3H/Heouj,Heouj组)小鼠复制部分肝脏缺血再灌注损伤模型,于缺血45min,再灌注1h和3h处死动物,检测血清天门冬氨酸氨基转移酶(AST)和血清肿瘤坏死因子α(TNFα)的含量;并以northern blot及髓过氧化物酶(MPO)试验分别检测缺血肝组织TNFα mRNA的表达和MPO的含量. 结果 (1)再灌注1、3h,与假手术组相比,小鼠血浆AST明显升高,但Hej组明显低于Heouj组(661.83U/L±106.09U/L和1215.5U/L±174.03U/L,t＝-6.65,P<0.01;1145.17U/L±132.43U/L和2958.17U/L±186.81U/L,t＝-5.57,P<0.01);(2)再灌注3h时,与假手术组相比,Hej组和Heouj组小鼠血清TNFα浓度明显升高,且前者明显低于后者(152.39pg/ml±43.3pg/ml和249.12pg/ml±51.89pg/ml,t＝-3.13,P<0.05);(3)再灌注1h,除假手术组外,Hej组和Heouj组小鼠缺血肝组织内可见TNFα mRNA的表达,但前者的表达水平明显低于后者,杂交带密度分析显示两者之间差异有显著性 (80.3±28.8与189.4±24.6,t＝-3.25,P<0.05);(4)再灌注3h,与假手术组相比,Hej组和Heouj组小鼠缺血肝组织内MPO含量明显升高,且前者含量明显低于后者(0.059±0.004和0.173±0.025,F＝33.49,P<0.01). 结论 Toll样受体可能通过其介导的炎性通路参与了小鼠肝脏缺血再灌注损伤.     

Candida-specific interferon-gamma deficiency and toll-like receptor polymorphisms in patients with chronic mucocutaneous candidiasis

Chronic mucocutaneous candidiasis (CMC) is a group of disorders, characterised by persistent mucocutaneous infections with Candida species. The underlying defect of CMC has not been elucidated, but a defective cytokine response may be involved. Therefore, we investigated whether an imbalance between IFNgamma and IL-10 may play a role in this disorder. We assessed the cytokine production in whole-blood cultures from CMC patients using Candida albicans, lipopolysaccharide and phytohaemagglutinin as stimuli. As the Toll-like receptors are important pattern recognition receptors for Candida species, we also investigated Toll-like receptor polymorphisms in these patients. Patients with CMC had a significantly decreased IFNgamma production when whole blood was stimulated with C. albicans (232 +/- 120 vs 2279 +/- 609 pg/ml, p<0.02). When stimulated with phytohaemagglutinin, the differences were not significant (3549 +/- 1320 vs 7631 +/- 1790 pg/ml). The Candida-stimulated production of IL-10 tended to be higher in CMC patients, whereas TNF and IL-1beta production were similar in patients and controls. Stimulation with LPS showed no differences in cytokine production between patients and controls. Two out of seven patients had the TLR4 Asp299Gly polymorphism and none had the TLR2 Arg677Trp polymorphism. These data support the hypothesis that deficient IFNgamma production is involved in the pathogenesis of CMC, whereas a role for genetic polymorphisms of Toll-like receptor 2 and 4 is not obvious in these patients.     

Differential activation of cytokine secretion in primary human colonic fibroblast/myofibroblast cultures

BACKGROUND: Fibroblasts and myofibroblasts are known to secrete a wide spectrum of cytokines, but the individual spectrum is tissue-specific. We investigated the effect of cell activation on cytokine secretion of isolated human colonic fibroblasts/myofibroblasts from control patients and patients with mucosal inflammation. METHODS: Primary cultures of human colonic submucosal fibroblasts/myofibroblasts were incubated with IL-1alpha (100 U/ml), IL-Ibeta (10 ng/ml), IL-10 (10 ng/ml), TNF (10 ng/ml), PMA (10 ng/ml), LPS (50 ng/ml), IL-4 (10 ng/ml), or a combination of IL-1 and TNF. Secreted cytokines were determined by ELISA. NF-kappaB activation was demonstrated by electrophoretic mobility-shift assays (EMSA). RESULTS: Incubation of colonic fibroblasts/myofibroblasts with IL-1, LPS, TNF and PMA induced secretion of IL-6, IL-8, M-CSF and GM-CSF. IL-8 and IL-6 secretion could be stimulated by IL-1alpha, IL-1beta, TNF, PMA and LPS within 6 h of incubation. IL-6 secretion was stimulated from 0.5 +/- 0.01 pg/h x microg fibroblast protein to 18.5 +/- 2.6 pg/h x microg fibroblast protein with IL-1beta (P < 0.01). IL-8 secretion was stimulated from 1.0 +/- 0.1 pg/h x microg fibroblast protein to 41.1 +/- 3.6 pg/h x microg (P < 0.005). IL-4 and IL-10 did not change cytokine secretion significantly. No significant differences between cultures from normal and inflamed mucosa were observed. TNF and IL-1 induced NF-kappaB activation. ALLN, a proteasome and NF-kappaB activation inhibitor, reduced TNF-mediated IL-8, GM-CSF and M-CSF induction significantly, whereas induction of IL-6 secretion remained unchanged. CONCLUSION: Human colonic myofibroblasts can secrete large amounts of IL-6, IL-8, M-CSF and GM-CSF upon stimulation. The induction of IL-8, M-CSF and GM-CSF, but not of IL-6 secretion, is mediated mainly by NF-kappaB activation. The cytokine profile and the total amounts of cytokines released suggest that colonic myofibroblasts can play a role in leukocyte recruitment and during mucosal inflammation. They therefore have to be regarded as an important part of the mucosal immune system.     

The effect of immunotherapy on interleukin-1 and tumor necrosis factor production of monocytes in asthmatic children.

A longitudinal study was conducted to determine the pathogenesis and effect of immunotherapy (IT) on monocyte function. Production of interleukin-1 (IL-1) and tumor necrosis factor (TNF) by peripheral blood monocytes in 31 asthmatic children before and one year after IT was compared. Twenty-two children completed the treatment course, and 13 age-matched healthy children served as controls. Adherent monocytes were isolated and stimulated with either crude mite extract of Dermatophagoid farinae (Df) for 7 days or lipopolysaccharide (LPS) for 3 days. The amount of TNF and IL-1 in culture supernatant was quantified by TNF and IL-1 enzyme-linked immunosorbent assay (ELISA) kits, respectively. The LPS-stimulated TNF production in patients was not different before or after IT (245.8 +/- 110.9 vs. 213.3 +/- 161.6 pg/0.1 ml, p +/- 0.202), but was significantly higher than the control (66.7 +/- 42.7 pg/0.1 ml; p less than 0.0001). The LPS-stimulated IL-1 production was similar among the three groups. When stimulated with Df antigen, monocytes from asthmatic patients produced a greater amount of TNF and IL-1 than did those from the control (p less than 0.001). Furthermore, although the production of TNF decreased after successful IT (360.2 +/- 181.6 vs 243.9 +/- 189.1 pg/0.1 ml, p less than 0.05), the production of IL-1 did not change (679.9 +/- 254.1 vs. 534.8 +/- 257.6 pg/0.1 ml, p greater than 0.05). Thus, repeated long-term administration of allergen (IT) was able to suppress specifically the TNF, but not IL-1 production of monocytes.(ABSTRACT TRUNCATED AT 250 WORDS)     

Tumor necrosis factor (cachectin) in human visceral leishmaniasis

High tumor necrosis factor-alpha (TNF alpha) levels were present in the serum of 24 of 28 active visceral leishmaniasis (VL) patients (142.9 +/- 113.9 pg/ml, mean +/- SD), whereas levels were not elevated in 26 of 30 patients with cryptic leishmanial infection (16 asymptomatic, 4 with self-healing subclinical infection, and 10 posttreatment VL cases). Serum TNF alpha levels were also not elevated in 15 normal volunteers (11.3 +/- 15.6 pg/ml) and in 10 patients with tegumentary leishmaniasis (19.1 +/- 10.8 pg/ml). Leishmanial infection of human monocyte-derived macrophages enhanced the basal TNF alpha production by these cells, and this effect was further potentiated by treatment with recombinant interferon-gamma. After effective treatment of VL patients, serum TNF alpha levels dropped rapidly (129 +/- 112 vs. 9 +/- 13 pg/ml in 10 days), even before clinical parameters such as spleen size or parasitism, white blood cell count, or levels of hemoglobin returned to normal values. On the other hand, patients unresponsive to treatment remained with elevated levels (276 +/- 69 vs. 155 +/- 71 pg/ml in 10 days). Thus, serum TNF alpha levels in VL patients are a good parameter to monitor in determining host response to therapy.     

The selected pro- and anti-inflammatory cytokines in the patients with coronary heart disease: preliminary communication

Recent findings suggest that inflammation and cytokines regulation may play a role in the pathogenesis of atherosclerosis and coronary heart disease. The aim of this study was to assess serum concentrations of selected pro- (TNF alpha) and antiinflammatory (IL-10) cytokines in patients with coronary heart disease. We studied 29 patients with coronary heart disease: 14 with stable angina (group I) and 15 with unstable angina (group II). The control group (group K) consisted of 10 healthy subjects. Patients with inflammatory diseases, previous myocardial infarction (last 6 months) and with ECG abnormalities, that would invalidate ST-segment analysis, were excluded from examined groups. We evaluated: clinical state of patients and results of some diagnostic examinations (lipids, ECG, echocardiography, coronary angiography, concomitant diseases). In each patients serum levels of TNF alpha and IL-10 were measured according to the special protocol by ELISA. The mean serum concentrations of TNF alpha and IL-10 were significantly higher in group I (respectively: 18.75 +/- 11.7 pg/ml, 89.0 +/- 114.9 pg/ml) and II (14.21 +/- 5.9 pg/ml, 49.38 +/- 72.9 pg/ml) in comparison to the healthy subjects (9.41 +/- 1.7 pg/ml, 9.69 +/- 4.5 pg/ml). We found positive correlations between mean TNF alpha and IL-10 concentrations in group II (48 hours after last symptom) and between mean TNF alpha concentration and LVM (left ventricular mass), LVMI (left ventricular mass index) in group I. The concentrations of TNF alpha and IL-10 did not correlate with other clinical parameters. The results of our study suggest that serum concentrations of pro- (TNF alpha) and antiinflammatory (IL-10) cytokines may be increased in patients with stable and unstable angina. These increased concentrations do not reflect the clinical state of patients.     

Polymicrobial sepsis selectively activates peritoneal but not alveolar macrophages to release inflammatory mediators (interleukins-1 and -6 and tumor necrosis factor).

While a number of clinical studies indicate that elevated serum cytokine [interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor (TNF)] levels are associated with enhanced mortality in sepsis, the time course and the role that different macrophage (M phi) populations play in releasing these cytokines remain to be determined. To study this, polymicrobial sepsis was induced in C3H/HeN mice by cecal ligation and puncture (CLP). The animals were then sacrificed at 1, 4, or 24 hr post-CLP. Blood was taken for serum cytokine level determination. Macrophages, of either peritoneal (PM phi) or alveolar (AM phi) origin, were harvested by lavage, and their innate vs. inducible cytokine productive capacities were assessed by incubation with or without endotoxin (lipopolysaccharide; LPS). Serum levels of TNF were significantly enhanced 1 hr post-CLP (CLP = 3.8 +/- 2.4* vs. sham = 0.4 +/- 0.9 U/ml; P less than 0.05 by t test). However, not until 4 hr post-CLP were marked increases in IL-6 observed (CLP = 318.0 +/- 209.0* vs. sham = 1.1 +/- 0.5 U/ml), which remained elevated through 24 hr post-CLP (CLP = 11.3 +/- 15.0* vs. sham = 0.03 +/- 0.02 U/ml). Cytokine release (IL-1, IL-6, TNF) from PM phi (without the addition of LPS) was detectable only in cells harvested 1 h following CLP. Alveolar M phi from septic mice showed little in vivo activation. Septic PM phi IL-1 and IL-6 production was markedly depressed at all time points with LPS stimulation, but TNF release remained unaltered.(ABSTRACT TRUNCATED AT 250 WORDS)