多器官功能障碍综合征血清炎性细胞因子变化的意义

目的 :探讨多器官功能障碍综合征 (MODS)血清炎性细胞因子的变化的意义。方法 :采用双抗体夹心酶联免疫吸附法测定 42例MODS和 3 0例健康对照组的血清肿瘤坏死因子 (TNF α)、白介素 1β(IL 1β)与白介素 6(IL 6)含量。结果 :MODS血清TNF α、IL 1β与IL 6含量 (各为2 77.64±5 4.3 6ng/L ,2 40 .97± 2 0 .87ng/L ,3 84.96± 73 .19ng/L)明显高于对照组 ( 2 3 .3 7± 7.96ng/L ,40 .65± 5 .87ng/L ,3 0 .2 6± 3 .61ng/L) ,P均 <0 .0 0 1;死亡组TNF α、IL 1β与IL 6含量 (各为5 5 4.86± 95 .69ng/L ,3 3 8.87± 41.2 1ng/L ,5 97.3 7± 118.3 6ng/L)高于非死亡组TNF α、IL 1β与IL 6含量 ( 2 77.64± 5 4.3 6ng/L ,2 40 .97± 2 0 .87ng/L ,3 84.96± 73 .19ng/L) ,P均 <0 .0 1。结论 :TNF α、IL 1β与IL 6对MODS的病理生理过程可能起作用 ,监测MODS患者血清TNF α、IL 1β与IL 6水平可作为反映病情严重程度和评估预后的一项参考指标     

白细胞介素7和15对肺结核患者Th1/Th2平衡的调节作用

目的探讨白细胞介素7(IL7)、IL15对肺结核病患者外周血单个核细胞(PBMC)分泌Th1型细胞因子γ干扰素(IFNγ)、肿瘤坏死因子α(TNFα)和Th2型细胞因子IL4、IL10的影响。方法选择2003年1至9月入院的60例肺结核患者和25名健康对照者,用葡聚糖泛影葡胺密度梯度离心法分离PBMC。按加入刺激物的不同,将每份标本分为6组:RPMI1640组、纯化蛋白衍生物(PPD组)、PPD+IL7组、PPD+IL7抗体组、PPD+IL15组、PPD+IL15抗体组。加入相应刺激物后培养72h,收集上清液,采用酶联免疫吸附法(ELISA)检测各组培养上清液中IFNγ、TNFα、IL4、IL10的水平。结果与PPD组相比,加入IL7的患者组PBMC分泌IFNγ和TNFα显著增高,分别为(107±42)～(157±74)ng/L、(460±128)～(887±242)ng/L;显著抑制IL4和IL10的合成,分别为(58±15)～(31±9)ng/L、(153±40)～(112±32)ng/L。健康对照组PBMC分泌IFNγ和TNFα显著增高,分别为(211±57)～(292±92)ng/L、(1203±390)～(1722±503)ng/L;显著抑制IL4和IL10的合成,分别为(43±13)～(36±11)ng/L、(135±37)～(96±36)ng/L。加入IL15患者组PBMC分泌IFNγ和TNFα显著增高,分别为(107±42)～(231±62)ng/L、(460±128)～(843±208)ng/L;显著抑制IL4和IL10的合成,分别为(58±15)～(37±9)ng/L、(153±40)～(116±41)ng/L。健康对照组PBMC分泌IFNγ和TNFα显著增高,分别为(211±57)～(343±108)ng/L、(1203±390)～(1468±235)ng/L;显著抑制IL4和IL10的合成,分别为(43±13)～(36±8)ng/L、(135±37)～(90±35)ng/L。加入IL7抗体或IL15抗体均可抑制IFNγ和TNFα的分泌,促进IL4和IL10的合成。肺结核患者各组IFNγ、TNFα水平均低于健康对照各组,而IL4、IL10水平比较差异无统计学意义。结论IL7和IL15可作为免疫调节剂,诱导IFNγ及TNFα分泌,抑制IL4及IL10合成,从而调节Th1/Th2平衡,发挥对结核分枝杆菌感染患者的免疫保护作用。     

哮喘患者白细胞介素(IL)-12和IL-13的变化及糖皮质激素对其的影响

目的　观察哮喘患者血清中白细胞介素 (IL) 12和IL 13水平的变化及糖皮质激素对其的影响。方法　采用ELISA法分别检测中度哮喘急性发作期患者 (2 5例 )口服泼尼松治疗 1周前后、缓解期患者 (2 0例 )和健康对照组者 (15例 )血清中IL 12和IL 13水平 ,并同时测 1秒钟用力呼气容积(FEV1)占预计值的百分比和气道阻力 (R5)占预计值的百分比。结果　IL 12水平急性发作期治疗前[(5 8 5± 14 2 )ng/L]较缓解期 [(71 3± 16 2 )ng/L]为低 (P <0 0 5 ) ,与健康对照组 [(85 5± 13 1)ng/L]、急性发作期治疗后 [(79 3± 19 1)ng/L]比较 ,差异均有显著性 (P <0 0 1)。IL 13水平急性发作期治疗前 [(131 3± 2 8 4 )ng/L]较缓解期 [(113 1± 2 6 5 )ng/L]为高 (P <0 0 5 ) ,与健康对照组 [(92 3± 14 4 )ng/L]、急性发作期治疗后 [(84 1± 19 8)ng/L]比较 ,差异均有显著性 (P <0 0 1)。FEV1占预计值的百分比下降和R5占预计值的百分比升高 (P <0 0 1)。直线相关分析表明 ,血清中IL 12与FEV1占预计值的百分比呈正相关 (r=0 4 85 ,P <0 0 5 ) ,与R5占预计值的百分比呈负相关 (r =- 0 5 16 ,P<0 0 5 ) ,与IL 13呈负相关 (r =- 0 5 4 9,P <0 0 1) ;IL 13与FEV1占预计值的百分比呈负相关 (r =- 0 4 93,P <     

不卧床持续性腹膜透析患者血浆和透析排出液中细胞因子的变化及意义

目的 :研究不卧床持续性腹膜透析 (CAPD)患者血浆和腹膜透析排出液中细胞因子的水平与腹膜透析和机体防御功能的关系。　 方法 :用酶免疫法测定 2 1例非感染期CAPD患者血浆和腹膜透析排出液中IL 1α、IL 1β、IL 6、TNFα、IFNγ和可溶性肿瘤坏死因子受体 1(sTNFR1)水平 ,同时做 4h腹膜平衡试验。比较细胞因子水平与腹透情况和腹膜转运特性的关系 ;另外 ,与同期测定的正常人血浆IL 6和sTNFR1水平作对比研究。　 结果 :2 1例CAPD患者血浆和腹透液中各种细胞因子水平分别为 :IL 1α 34 40± 12 87ng/L和 17 6 0± 10 49ng/L(P <0 0 0 1) ;IL 6 8 10± 14 6 9ng/L和 15 7 6 5± 130 2 3ng/L(P <0 0 0 1) ;TNFα 117 30± 195 2 7ng/L和 2 2 90± 13 37ng/L(P <0 0 5 ) ;sTNFR19 76± 0 98μg/L和 1 84± 2 72 μg/L(P <0 0 0 1)。仅在 8例患者血浆及 9例患者腹透液中检测出IL 1β;在 1例患者血浆中测到IFNγ ,腹透液中IFNγ为 2 14± 0 74kU/L。腹透液中IL 6和IFNγ水平呈负相关 (P <0 0 2 )。血浆和腹透液中各种细胞因子水平与腹透时间长短、既往感染与否、透析充分性和超滤量以及腹膜转运特性均无关系。CAPD患者血浆IL 6和sTNFR1水平比正常人明显升高 (P <0 0 0 1)。　 结论 :腹透液中IL     

肝硬化患者血清—氧化氮和几种白细胞介素检测的临床意义

目的　探讨肝硬化患者血清一氧化氮 (NO)、白细胞介素 2 (IL 2 )、白细胞介素 6 (IL 6 )、白细胞介素 8(IL 8)及可溶性白细胞介素 2受体 (sIL 2R)变化的临床意义。方法　应用ELISA法测定肝硬化患者及正常对照组的血清IL 6、IL 8及sIL 2R含量 ;应用MTT法检测血IL 2活性 ;应用荧光法检测血清NO水平。结果　肝硬化患者血清IL 2、sIL 2R、IL 6、IL 8及NO水平 :(5 741.5 3± 4376 .5 2 )U/ml、(486 .76± 46 .41)U/ml、(15 .78± 3.0 4) pg/ml、(2 3.89± 2 .13)pg/ml及 (6 .33± 0 .37) μmol/L ,显著高于正常对照组 :(173.88± 92 .2 1)U/ml、(2 42 .36± 35 .78)U/ml、(6 .14± 3.12 ) pg/ml、(17.71± 1.32 )pg/ml及 (3.6 8± 0 .34 ) μmol/L ,并随肝功受损程度进行性增加。 结论　肝硬化患者血清IL 2、sIL 2R、IL 6及IL 8增加可能为NO增多的诱发因素 ;肝功能损伤可能是白细胞介素活性增加的重要原因。     

肝炎后肝硬化患者外周血IL-10、IL-12的水平及其意义

目的　探讨IL -10、IL -12在肝炎后肝硬化中的作用及意义。方法　 3 4例肝炎后肝硬化患者根据Child -Pugh分级法分为A、B、C级三组 ,应用ELISA法检测外周血IL -10、IL -12水平 ,10例健康体检者作对照。结果　 3 4例肝炎肝后硬化患者有2 0例HBVDNA(+ ) ,HBVDNA(+ )组患者IL -10水平明显高于HBVDNA(-)组 (4 3 2 9± 18 66pg/mL、2 1 42± 9 47pg/mL ,P <0 0 1) ,而IL -12水平则低于HBVDNA(-)组 (92 45 + 2 9 73pg/mL、2 0 7 3 4± 83 61pg/mL ,P <0 0 1) ;肝硬化患者A级者IL -10水平稍高于对照组 (4 5 2 6± 9 43pg/mL、3 7 42± 5 61pg/mL ,P <0 0 5 )但显著高于B级和C级 (2 0 89± 7 46pg/mL、9 13± 0 2 4pg/mL ,P <0 0 1) ,且C级又明显低于B级 (P <0 0 1) ;肝硬化A、B、C级患者IL -12水平均低于对照组 (12 4 2 7± 5 0 14pg/mL、10 6 42± 49 0 6pg/mL、70 3 6± 2 0 62pg/mL ,2 15 64± 78 3 7pg/mL ,P <0 0 1) ,而以C级患者最低 (P <0 0 1)。结论　IL -10、IL -12可能在肝炎后肝硬化的发生发展中有一定作用     

小肠细菌过度生长相关性轻微肝性脑病患者的血浆内毒素水平

目的观察肝硬化患者小肠细菌过度生长相关性轻微肝性脑病患者的内毒素水平,探讨内毒素与轻微肝性脑病的关系。方法对90例肝硬化患者及20例健康志愿者进行内毒素水平、葡萄糖氢呼气试验（GHBT）、数字连接试验（NCT—A和NCT—BC）和数字符号试验（DST）检测,观察小肠细菌过度乍长相关性轻微肝性脑病患者的内毒素水平变化。结果肝硬化组小肠细菌过度生长（36．7％,33／90）明显高于健康对照组（5％,1／20）。20名健康志愿者未检出轻微肝性脑病（MHE）;90例肝硬化患者轻微肝性脑病37例（41．1％）,其中伴小肠细菌过度生长肝硬化患者轻微肝性脑病发生率高于不伴小肠细菌过度生长患者。肝硬化组内毒素水平（69．6±21．3）pg／mL高于健康对照组（18．7±8．6）pg／mL;肝硬化小肠细菌过度生长组内毒素水平（89．5±17．6）pg／mL高于无小肠细菌过度生长组（57．3±15．8）pg／mL（P〈0．05）。应用抗生素抑制小肠细菌过度生长及乳果糖治疗1周后GHBT、内毒素水平、NCT—A、NCT-BC及DST检查结果改善。结论内毒素与伴小肠细菌过度生长的轻微肝性脑病的发生及进展可能有一定的关系。     

心力衰竭患者炎性与抗炎性细胞因子表达的平衡失调

目的 :了解炎性与抗炎性细胞因子在充血性心力衰竭 (CHF)过程中的变化及其临床意义。方法 :用双抗体夹心ELISA法测定 12 2例CHF患者及 30例健康人血浆中肿瘤坏死因子 α(TNF α)、白细胞介素 6 (IL 6 )、白细胞介素 10 (IL 10 )的浓度。结果 :①CHF患者血浆中TNF α水平明显高于对照者 (P <0 .0 5或 <0 .0 1) ,且随着心力衰竭程度的加重 ,TNF α水平呈进行性增高 ;CHF患者血浆IL 6及IL 10水平 ,心功能Ⅲ、Ⅳ级者明显高于对照者 (P <0 .0 5或 <0 .0 1) ,而心功能Ⅱ级者与对照者相比差异无显著性意义。②TNF α与IL 6 (r =0 .6 18,P <0 .0 1)、IL 10 (r =0 .5 6 6 ,P <0 .0 1)均呈正相关 ,但TNF α与IL 10的比率 (TNF α/IL 10 )也随着心功能的恶化而升高 ,IL 10的升高与TNF α的升高相比明显不足。结论 :细胞因子的变化与心力衰竭的严重程度密切相关 ,CHF患者血中炎性细胞因子明显升高的同时伴有抗炎性细胞因子升高的相对不足 ,炎性与抗炎性细胞因子之间的平衡失调可能参与了CHF的发生发展     

老年慢性支气管炎患者血清细胞因子检测的意义

目的　探讨白细胞介素 2 (IL 2 )、IL 6、IL 8和肿瘤坏死因子 α(TNF α)在慢性支气管炎发病机制中的作用。　 方法 　采用放射免疫法 (RIA)检测 3 0例慢性支气管炎患者急性发作期和临床缓解期血清IL 2、IL 6,IL 8及TNF α水平 ,并以 3 0例健康人血清中IL 2、IL 6、IL 8及TNF α水平作为对照。　 结果 　 3 0例慢性支气管炎患者急性发作期血清IL 6、IL 8、TNF α显著高于缓解期 (P <0 0 1) ,而缓解期血清IL 6、IL 8、TNF α与对照组相比仍然有差异 (P <0 0 1) ;IL 2浓度在发作期和缓解期均低于对照组 (P <0 0 5 ) ,但发作期和缓解期比较差异无显著性。慢性支气管炎患者急性发作期和临床缓解期血清IL 6与IL 8,IL 6与TNF α ,IL 8与TNF α水平均呈正相关 (r=0 70 9,0 63 3 ,0 615 ) ,但与IL 2水平无明显相关性。　 结论 　血清IL 2、IL 6、IL 8、TNF α不仅参与了慢性支气管炎急性感染时的发病机制 ,而且在慢性炎症的发展过程中发挥重要作用     

慢性阻塞性肺疾病患者炎症细胞因子与肺通气功能的相关研究

目的　探讨慢性阻塞性肺疾病 (COPD)患者肺通气功能改变与炎症因子变化之间的关系。方法　稳定期COPD和慢性支气管炎 (简称慢支 )患者各 8例 ,,另有 8名健康者作为对照 ,进行肺功能检查 ,并经支气管肺泡灌洗获取肺泡巨噬细胞进行培养 ,采用酶联免疫吸附 (ELISA)方法测定大肠杆菌内毒素 (LPS)刺激后上清液中白细胞介素 8(IL 8)、IL 1β、IL 6和肿瘤坏死因子α(TNF α)的浓度 ,细胞因子之间相关性采用Pearson相关阵分析 ,肺功能值与细胞因子相关性采用多元后退回归法分析。结果　 (1)肺泡巨噬细胞释放IL 8:加入LPS后COPD组为 [(43± 2 7) μg/L和 (5 7± 41) μg/L],与正常对照组 [(13± 10 ) μg/L和 (2 0± 13 ) μg/L) ]比较差异有显著性 (P <0 .0 5 ) ;与慢支组 [(2 9± 2 1)μg/L和 (3 2± 2 3 ) μg/L]比较差异有显著性 (P >0 .0 5 )。 (2 )加入LPS前、后 ,COPD组、慢支组和正常对照组肺泡巨噬细胞释放IL 1β分别为 [(5 0± 41)ng/L、(94± 5 9)ng/L、(3 7± 3 2 )ng/L、(2 2 5± 10 8)ng/L、(15 3± 175 )ng/L、(70± 3 7)ng/L],与IL 8的释放呈正相关 (P <0 .0 5 ) ;三组肺泡巨噬细胞在LPS刺激后释放TNF α分别为 [(12 3 8± 679)ng/L、(3 0 88± 2 879)ng/L、(13 3 2± 1846)ng/L],与IL 1β呈正相     

Use of the lactose-[13C]ureide breath test for diagnosis of small bowel bacterial overgrowth: comparison to the glucose hydrogen breath test

Purpose  

The glucose hydrogen breath test (GHBT) is commonly used as a noninvasive test to diagnose small bowel bacterial overgrowth (SBBO) but its validity has been questioned. Our aim was to evaluate the lactose-[¹³C]ureide breath test (LUBT) to diagnose SBBO and to compare it with the GHBT, using cultures of intestinal aspirates as a gold standard.     

Intestinal damage mediated by Kupffer cells in rats with endotoxemia

AIM：To determine the in vivo effects of phagocytic blockade of Kupffer cell(KC)on the release of proinflammatory cytokines in small intestinal lesion and on the integrity of intestinal tract by using gadolinium chloride(GdCl3)during early endotoxemia.METHODS:Wistar rats were divided into three groups:GropA,rats were injected with endotoxin(E.coliO111:B4,adose of 12mg&#183;kg^-1)only;GroupB,rats were pretreated intravenously with 25mg of GdCl3per kg24hare given endotoxin;and Group C,sham operation only.All animals were sacrificed 4h after endotoxin injecton.In portion of the rats of three groups.bile duct was cannulated,which the bile was collected externally.Morphological changes of ileum were observed under light microscopy and electronic microscopy.The KC were isolated rfrom rats by collagenase perfusion and inKC,expression of TNF-αand IL-6mRNA were determined by RT-PCRanalysis.Plasma and bile TNF-αandIL-6Levels were determined by enzyme-linked immunosorbent assay(EISA)&gt;RESULTS:In group A,there were neutrophil infiltration and superficial epitelial necrosis of the ilealvvilli,sloughing of mucosal epithelium.and disappear ance of somevilli.In groupB.the ileal mucosal damage was much reduced.which in groupC,no significant morphological changes were seen,GdCl3pretreatment decreased significantly the expression of TNF-αand IL-6mRNA ingroupB(4.32&#177;0.47and4.05&#177;0.43)when compared to groupA(9.46&#177;1.21and9.04&#177;1.09)(P&lt;0.05).There was no significant expression of TNF-αand IL-6mRNA in groupC(1.03&#177;0.14and10.4&#177;0.13).In rats of groupA,the levels of TNF-αand IL-6in bile and plasma were 207&#177;29ng&#183;L^-1,1032&#177;107ng&#183;L^-1,213&#177;33ng&#183;L^-1,and 1185&#177;127ng&#183;L^-1,respectively.In groupB,they were113&#177;18ng&#183;L^-1,repectively.In groupC,they were 67&#177;10ng&#183;L^-1,72&#177;13ng&#183;L^-1,109&#177;18ng&#183;L^-1,and 118&#177;22ng&#183;L^-1respectively.There were significant difference between the three group(P&lt;0.05).CONCLUSION:KC release cytokinesTNF-αand IL-6causing damage to the integrity of intestinal epithelium and play a crucial role in the initiation and progression of intestinal mucosal damage during early endotoxemia.     

Pretreatment with ibuprofen augments circulating tumor necrosis factor-alpha, interleukin-6, and elastase during acute endotoxinemia

Plasma levels of tumor necrosis factor-alpha (TNF alpha), interleukin-1 (IL-1), and interleukin-6 (IL-6) were monitored after intravenous administration of Escherichia coli endotoxin with or without ibuprofen pretreatment to healthy volunteers. Intravenous endotoxin (n = 7) resulted in elevated plasma TNF alpha concentrations with maximal levels at 90 min (369 +/- 44 pg/ml, P less than .001 vs. saline controls, n = 7). The rise in TNF-alpha was followed by a rise in plasma IL-6 (27 +/- 12.8 ng/ml), peaking 30-90 min thereafter. Pretreatment with ibuprofen (n = 6) caused a significant augmentation and temporal shift in cytokine elaboration with maximal TNF alpha levels (627 +/- 136 pg/ml) at 120 min and IL-6 peaks (113 +/- 66 ng/ml) at 180 min. In ibuprofen-treated volunteers, the additional increase in TNF alpha was paralleled by increased levels of circulating elastase. In vitro experiments suggest a causal relationship between these events. Thus, the cyclooxygenase inhibitor ibuprofen blunts the clinical response to endotoxin but augments circulating cytokine levels and leukocyte degranulation.     

Circulating interleukin-1 and tumor necrosis factor in septic shock and experimental endotoxin fever

Interleukins (IL) -1 beta and -1 alpha and tumor necrosis factor (TNF-alpha) were measured by radioimmunoassay in plasma samples from 44 healthy individuals, 15 patients in septic shock, and 6 volunteers infused with endotoxin. Plasma IL-1 alpha levels were low (40 pg/ml) or undetectable in all situations. In 67% of the healthy subjects, plasma IL-1 beta levels were less than 70 pg/ml. Septic patients had higher plasma IL-1 beta levels (120 +/- 17 pg/ml, P = .001); those of surviving patients were higher than those of patients who died (P = .05). Plasma TNF-alpha concentrations in septic individuals were elevated (119 +/- 30 pg/ml) and correlated with severity of illness (r = .73, P = .003), but no correlation was observed between plasma IL-1 beta and TNF-alpha concentrations in individual samples. Infusion of endotoxin caused a twofold elevation of IL-1 beta, from a baseline of 35 +/- 5 pg/ml to a maximum of 69 +/- 27 pg/ml at 180 min (P less than .05). Peak TNF-alpha levels after endotoxin infusion were 15 times higher than IL-1 beta levels, were attained more rapidly (90 min), and as with the septic patients, did not correlate with IL-1 beta levels. These data support the concept that plasma IL-1 beta and TNF-alpha concentrations are regulated independently and are associated with different clinical outcomes.     

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)     

Relationship between the tumor necrosis factor system and the serum interleukin-4, interleukin-5, interleukin-8, eosinophil cationic protein, and immunoglobulin E levels in the bronchial hyperreactivity of adults and their children.

The aim of the study was to investigate the activation of inflammatory mediators interleukin (IL)-4, IL-5, and IL-8; immunoglobulin E (IgE); and eosinophil cationic protein (ECP) and to evaluate the regulatory role of the tumor necrosis system (TNF) system in bronchial hyperreactivity. Adults who had suffered from bronchial asthma in childhood but who had been symptom free for at least 3 years were examined together with their children who did not have asthma. The serum concentrations of TNF-alpha, soluble TNF receptor 1 (sTNF-R1), TNF-R2, IL-4, IL-5, IL-8, ECP, and IgE were studied in symptom-free adults (n = 22) and their children (n = 22) with bronchial hyperreactivity. Nonhyperreactive individuals with a similar medical history (adults, n = 17; children, n = 20) served as controls. Significantly elevated serum TNF-alpha (X +/- SD: 5.13 +/- 1.37 pg/mL versus 3.91 +/- 0.61 pg/mL; p < 0.0001), sTNF-R1 (X +/- SD: 1.37 +/- 0.28 ng/mL versus 1.16 +/- 0.13 ng/mL; p = 0.0002), and sTNF-R2 (X +/- SD: 0.78 +/- 0.42 ng/mL versus 0.43 +/- 0.41 ng/mL; p = 0.0001); IL-4 (X +/- SD: 4.05 +/- 1.02 pg/mL versus 3.34 +/- 0.84 pg/mL; p = 0.0016); IgE (X +/- SD: 390.1 +/- 361.4 KU/L versus 130.2 +/- 166.1 KU/L; p = 0.0001); and ECP (X +/- SD: 17.57 +/- 11.03 micrograms/L versus 10.65 +/- 6.01 micrograms/L; p = 0.0016) concentrations were measured in the subjects with bronchial hyperreactivity as compared with the nonhyperreactive group. Significant positive linear correlations were observed for the bronchial hyperreactive group between the concentrations of TNF-alpha and ECP, TNF-alpha and sTNF-R1, TNF-alpha and IL-8, sTNF-R1 and ECP, sTNF-R1 and IL-8, and sTNF-R2 and IL-8. Moreover, the TNF-alpha and sTNF-R2 levels correlated with the airway reactivity in the hyperreactive group. We suggest that the elevated cytokine levels indicate activation of the immune system in individuals who were previously asthmatic, but recovered, and are now symptom free and in their children with nonasthmatic bronchial hyperreactivity. The TNF system may play a key role in the pathomechanism of bronchial hyperreactivity.     

Analysis of circulating apoptosis mediators and proinflammatory cytokines in patients with idiopathic hypertrophic cardiomyopathy: comparison between nonobstructive and dilated-phase hypertrophic cardiomyopathy

We examined the plasma levels of soluble Fas (sFas) or Fas ligand (sFas-L), tumor necrosis factor-alpha (TNF-alpha), and interleukin-6 (IL-6) in patients with idiopathic nonobstructive (HNCM) and dilated-phase (DHCM) hypertrophic cardiomyopathy. Patients with idiopathic hypertrophic cardiomyopathy (HCM) may deteriorate to DHCM and the pathogenesis is unknown. The levels of these plasma cytokines were measured by ELISA and echocardiography was performed in 38 HNCM and 11 DHCM patients, and 10 normal subjects. The follow-up period was three years. In HNCM, TNF-alpha (43.3 +/- 45.2 versus 16.9 +/- 4.3 pg/mL) and IL-6 (65.1 +/- 86.4 versus 4.0 +/- 2.1 pg/mL) were slightly higher compared to normal subjects and sFas (3.7 +/- 1.2 versus 2.1 +/- 0.7 ng/mL) increased significantly. sFas (3.9 +/- 1.8), TNF-alpha (79.3 +/- 72.4), and IL-6 (234.1 +/- 135.2) in DHCM were significantly increased and only IL-6 was significantly different from HNCM. sFas-L (0.18 +/- 0.08 versus 0.25 +/- 0.05 ng/mL) in HNCM was significantly decreased, and the decrease was marked in DHCM (0.05 +/- 0.02). In HNCM, TNF-alpha was negatively correlated with fractional shortening (r = -0.432, P = 0.0062) or positively with IL-6 (r = 0.665, P < 0.0001), while sFas-L was negatively correlated with IL-6 (r = -0.580, P < 0.0001). DHCM with high sFas had significantly higher cumulative incidences of worsening heart failure. The Fas/Fas-L system and proinflammatory cytokines may play an important role in the status of HCM and its progression to DHCM.     

Pathophysiologic quantities of endotoxin-induced tumor necrosis factor-alpha release in whole blood from patients with chronic heart failure

Bacterial endotoxin activity is elevated in patients with decompensated chronic heart failure (HF) and acts as a potent stimulus for immune activation. We sought to determine whether endotoxin, at an activity level seen in vivo (around 0.6 EU/ml), is sufficient to stimulate the secretion of tumor necrosis factor-alpha (TNF-alpha) and TNF-alpha soluble receptor (sTNFR2) in ex vivo whole blood from patients with HF. We studied 15 patients with HF (aged 65 +/- 1.9 years, New York Heart Association class 2.1 +/- 0.3, left ventricular ejection fraction 31 +/- 5%; mean +/- SEM), of whom 5 had cardiac cachexia, and 7 healthy control subjects (59 +/- 5 years, p = NS). Reference endotoxin was added to venous blood at concentrations of 0.6, 1.0, and 3.0 EU/ml, and was incubated for 6 hours. Endotoxin induced a dose-dependent increase in TNF-alpha release (p <0.05 in all groups). Patients with noncachectic HF produced significantly more TNF-alpha compared with controls after stimulation with 0.6, 1.0, and 3.0 EU/ml of endotoxin (113 +/- 46 vs 22 +/- 4 [p = 0.009], 149 +/- 48 vs 34 +/- 4 [p = 0.002], and 328 +/- 88 vs 89 +/- 16 pg/ml [p = 0.002], respectively; mean +/- SEM). Patients with cardiac cachexia produced significantly less TNF-alpha compared with patients without cardiac cachexia for all given concentrations (all p <0.05, analysis of variance p = 0.02). Production of sTNFR2 was greater at all concentrations of endotoxin versus controls (all p <0.05, analysis of variance p = 0.002). Plasma endotoxin levels were higher in patients with cardiac cachexia (4.3 times higher than in control subjects, p <0.005). Thus, low endotoxin activity, at levels seen in vivo in patients with HF, induces significant TNF-alpha and sTNFR2 production ex vivo. These results suggest that elevated plasma endotoxin activity observed in patients with HF is of pathophysiologic relevance.     

In vitro plasma perfusion through adsorbents and plasma ultrafiltration to remove endotoxin and cytokines.

In vitro plasma perfusion experiments were performed using small columns containing either resin or charcoal adsorbents to assess the removal of cytokines and endotoxin. 125I-labelled tumor necrosis factor-alpha (TNF-alpha; 500 pg/ml) and interleukin-6 (IL-6; 10 ng/ml) were added individually to human plasma. Over 4 hr of perfusion, Amberlite XAD-7 resin removed 32.5% +/- 3.3% (n = 5) of the initial amount of TNF-alpha and 71.4% +/- 3.8% (n = 5) of the initial amount of IL-6. DHP-1 polyhema-coated activated charcoal removed 17.2% +/- 6.2% (n = 5) of TNF-alpha and 48.5% +/- 7.4% (n = 5) of IL-6. Preliminary experiments were performed with lipopolysaccharide (LPS; 100 ng/ml) and interleukin-1 alpha (IL-1 alpha; 500 pg/ml), which showed that, over 4 hr, Amberlite XAD-7 removed 10.3% of the initial LPS and 29.1% of IL-1 alpha, whereas DHP-1 charcoal removed 23.2% of the initial LPS and 65.3% of IL-1 alpha. In vitro plasma ultrafiltration with either polysulfone or polyacrylonitrile membranes, as used clinically in haemodialysis, was performed with recirculation of plasma containing LPS or TNF-alpha. Neither of the substances was filtered to a significant degree. In conclusion, direct removal of these inflammatory mediators from the circulation of patients with multiorgan failure due to fulminant hepatic failure or sepsis would be possible by perfusion of plasma through adsorbents but not by haemodialysis.