重组人干扰素α-2b对慢性乙型肝炎患者T淋巴细胞免疫效应分子表达的影响

[目的]探讨重组人干扰素α-2b对慢性乙型肝炎(CHB)患者T淋巴细胞免疫效应分子表达的影响。[方法]选取我院2015-06—2016-01收治的CHB患者102例。根据患者治疗药物的不同,将102例患者分为对照组及观察组,每组患者51例。另外选取同时期我院15例健康体检者作为正常对照组。对照组患者采用恩替卡韦进行治疗,观察组采用干扰素α-2b联合恩替卡韦进行治疗。比较患者治疗前后外周血T细胞的免疫效应分子穿孔素(PE)、颗粒酶B(GrB)、颗粒溶素(GNLY)、肿瘤坏死因子-α(TNF-α)、γ-干扰素(IFN-γ)的表达情况。[结果]治疗前后CD_4~+ T细胞免疫效应分子表达水平变化:对照组、观察组患者治疗前后PE、GrB、GNLY、TNF-α、IFN-γ水平与正常对照组比较,均存在明显差异(均P0.05)。治疗后,观察组GNLY、TNF-α、IFN-γ水平显著低于对照组(P0.05);观察组患者治疗后GrB、GNLY、TNF-α、IFN-γ水平较治疗前均显著下降(P0.05),而对照组患者治疗后仅GrB、IFN-γ水平较治疗前显著下降(P0.05)。治疗前后CD_8~+ T细胞免疫效应分子表达水平变化情况:对照组、观察组患者治疗前后PE、GrB、GNLY、TNF-α、IFN-γ与正常对照组比较,均存在明显差异(均P0.05)。对照组、观察组患者治疗后PE、GNLY、TNF-α、IFN-γ均较治疗前明显下降(P0.05),其中观察组治疗后GNLY、TNF-α下降幅度明显高于对照组(P0.05)。[结论]重组人干扰素α-2b联合恩替卡韦可有效降低T淋巴细胞免疫效应分子的表达水平。     

早期特发性膜性肾病患者血清IL-2、IL-10、TNF-α和IFN-γ水平及临床意义

目的:探究早期特发性膜性肾病(IMN)患者血清白细胞介素-2(IL-2)、白细胞介素-10(IL-10)、肿瘤坏死因子-α(TNF-α)和干扰素-γ(IFN-γ)水平及临床意义。方法:选择2019年1月至2020年12月诊治的125例IMN患者作为观察组,另选取同时期100例健康体检者作为对照组。观察组治疗后33例未缓解,92例缓解。检测2组血清IL-2、IL-10、TNF-α和IFN-γ水平及观察组治疗前、后其水平,分析观察组治疗前血清IL-2、IL-10、TNF-α和IFN-γ水平与各项临床指标的相关性,比较不同疗效患者血清IL-2、IL-10、TNF-α和IFN-γ水平的变化。结果:观察组血清IL-2、TNF-α水平较对照组高,血清IL-10、IFN-γ水平较对照组低(P<0.05);观察组血压、24 h尿蛋白、血清C反应蛋白(CRP)均较对照组高(P<0.05);Pearson相关性分析显示,观察组IL-10水平与血压呈负相关(P<0.05),TNF-α水平与24 h尿蛋白、CRP均呈正相关,IFN-γ水平与血压、CRP均呈负相关(P<0.05);缓解组...     

化疗前后肺癌患者血清IFN-γI、L-6、TNF-α水平变化及意义

目的观察化疗前后肺癌患者血清干扰素-γ(IFN-γ)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)的水平变化,并探讨其临床意义。方法对60例肺癌患者(观察组)化疗前后及30例健康查体者(对照组)血清中的IFN-γI、L-6和TNF-α采用双抗体夹心ELISA法测定。结果化疗后观察组血清IL-6、TNF-α水平均明显低于、血清IFN-γ水平明显高于化疗前及对照组(P均<0.05);观察组化疗后肺癌复发患者血清IFN-γI、L-6、TNF-α与化疗前比较,P均>0.05。结论化疗后,肺癌患者血清IL-6、TNF-α水平降低I、FN-γ水平升高;动态观这些指标有助于指导治疗、评价疗效和判断预后。     

原发性高血压病患者细胞因子水平与左心功能的相关分析

目的探讨原发性高血压病(EH)患者血清中细胞因子水平与左心功能的关系.方法选择58例EH患者按超声心动图左室重量指数(LVMI)分为左室肥厚组及非左室肥厚组,均行EF(左室射血分数)值、血压值(BP)、血清IL-1、IL-6、TNF-α、IFN-γ的检测,并与30例正常对照组比较.结果EH组患者血清IL-1、IL-6、TNF-α明显高于对照组(P＜0.01),IFN-γ、EF、E/A明显低于对照组(P＜0.01).相关分析表明,左心功能指标EF、E/A值与IL-1、IL-6、TNF-α呈负相关,与IFN-γ呈正相关.结论高水平IL-1、IL-6、TNF-α及低水平IFN-γ对EH心功能损害的发生产生不利影响.     

HBeAg阳性慢性乙型肝炎患者肝功能损伤程度与Th1/Th2细胞因子水平的相关性研究

目的探讨HBeAg阳性慢性乙型肝炎患者肝功能损伤程度与Th1/Th2细胞因子水平的相关性。方法选择2012年1月至2016年12月来我院就诊的85例HBeAg阳性CHB患者及30名同期体检的健康者,根据ALT、TBil、A、A/G及凝血酶原活动度,将HBeAg阳性慢性乙型肝炎患者分为轻、中、重度三组。观察轻、中、重度组及对照组间的Th1/Th2细胞因子水平(IFN-γ、TNF-α、IL-4及IL-10),并观察IFN-γ、TNF-α、IL-4及IL-10分别与HBeAg阳性慢性乙型肝炎患者不同肝损伤程度的相关性。结果 IFN-γ与HBeAg阳性慢性乙型肝炎患者肝损伤程度呈正相关,IL-4及IL-10与其呈负相关,TNF-α与其无相关性。结论 Th1/Th2细胞因子相关水平(IFN-γ、IL-4及IL-10)可以反映HBeAg阳性慢性乙型肝炎患者的免疫状态,可作为病毒治疗时机选择的免疫学参照指标。     

慢性HBV感染者血清IL-15和外周血iNKT细胞水平变化及相关性研究

目的探讨慢性乙肝病毒(hepatitis B virus,HBV)感染者血清IL-15和外周血恒定自然杀伤T(invariant nature killer T,i NKT)细胞水平的变化。方法收集54例慢性HBV感染者和15名健康者外周血,分离血清和外周血单个核细胞(PBMC);ELISA法检测血清中IL-15水平;流式细胞术检测i NKT细胞占T细胞的比例,PMA、BFA和ionomycin处理PBMC后,流式细胞术检测i NKT分泌的IFN-γ和TNF-α水平,并利用Pearson法进行相关性分析。结果与正常对照者相比,免疫清除期患者血清中IL-15水平明显升高(P0.05),免疫耐受期患者高于正常对照者,低于免疫清除组,差异无统计学意义(P0.05);免疫清除期和免疫耐受期患者血清ALT水平和HBV DNA载量与IL-15水平无明显相关性(P0.05)。免疫清除期患者外周血i NKT细胞比例显著降低,免疫耐受期患者略升高,但差异无统计学意义(P0.05)。免疫耐受期和清除期患者外周血i NKT细胞分泌IFN-γ和TNF-α的细胞比例略有升高,但差异无统计学意义(P0.05)。免疫耐受期和免疫清除期患者i NKT细胞分泌IFN-γ水平与IL-15水平呈正相关;免疫耐受期和免疫清除期患者i NKT细胞分泌TNF-α水平与IL-15水平无相关性(P0.05)。结论慢性HBV感染者血清中IL-15水平升高,且外周血i NKT细胞数量减少,且不同感染状态患者血清IL-15水平和外周血i NKT细胞数量呈动态变化。     

莫西沙星联合鞘内注射及脑脊液置换治疗有效降低结核性脑膜炎患者脑脊液CK-BB、sCD163及炎症因子

目的:探讨莫西沙星联合鞘内注射及脑脊液置换治疗对结核性脑膜炎(TBM)患者脑脊液肌酸激酶同工酶BB(CK-BB)、可溶性CD163(s CD163)、炎症因子(TNF-α、IFN-γ)水平的影响及其临床疗效。方法:选取2015年4月-2019年4月,在河北大学附属医院结核科住院治疗的难治性TBM患者80例,将其按照随机数字表法分为对照组和观察组,各40例。2组患者均行鞘内注射、脑脊液置换治疗及抗结核基础治疗;观察组在此基础上予以莫西沙星静脉滴注治疗。比较2组患者脑脊液中CK-BB、s CD163、肿瘤坏死因子α(TNF-α)、γ干扰素(IFN-γ)含量变化及脑脊液生化指标,并进行疗效判断,观察2组不良反应发生情况。结果:治疗28 d后,2组患者脑脊液中CK-BB、s CD163及TNF-α、IFN-γ白细胞、蛋白、脑脊液压力显著降低,且观察组低于对照组(P均0. 05);观察组治疗总有效率高于对照组(P 0. 05)。与治疗前比较,2组患者脑脊液中葡萄糖、氯化物水平显著提高,且观察组高于对照组(P均0. 05),不良反应发生率无明显差异。结论:莫西沙星联合鞘内注射及脑脊液置换治疗能有效降低TBM患者脑脊液CK-BB、s CD163、炎性因子TNF-α、IFN-γ、白细胞及蛋白含量,提高葡萄糖和氯化物水平,且临床疗效较好。     

光动力对尖锐湿疣患者外周血自然杀伤T细胞及相关细胞因子的影响

目的探讨光动力(ALA-PDT)治疗对尖锐湿疣(CA)患者外周血自然杀伤T细胞(NKT)比值及细胞因子IL-4和干扰素IFN-γ浓度的影响。方法 124例CA患者在第1次ALA-PDT治疗前及连续3次ALA-PDT治疗后分别用流式细胞仪进行外周血NKT细胞比值及细胞因子IL-4和干扰素IFN-γ浓度的测定,比较各组数值之间的差异,并观察皮疹消退情况。20例健康体检者作为正常对照组。结果 ALA-PDT治疗前CA患者组外周血NKT细胞比值及细胞因子IL-4和IFN-γ浓度明显低于健康对照组,且复发组明显低于初发组(P均0.001)。治疗后CA患者组3项指标随治疗次数增加逐渐升高(P均0.01);3次治疗后总CA组NKT细胞比值、IL-4浓度仍低于健康对照组(P均0.001),但IFN-γ浓度与健康对照组差异无统计学意义(P0.05),初发组3项指标与正常对照组差异无统计学意义(P均 0.05),但复发组3项指标仍低于健康对照组(P均0.001),且复发组NKT细胞比值、IL-4浓度仍低于初发组(P均0.001),但IFN-γ浓度与初发组差异无统计学意义(P0.05)。3次治疗后皮疹未完全消退组3项指标仍低于消退组(P均0.001),同时低于健康对照组(P均0.001),但皮疹完全消退组3项指标不再低于健康对照组(P均0.05)。3次ALA-PDT治疗后疣体全部消退患者占比70.97%。结论 CA发病可能引起患者外周血NKT细胞比值减少和细胞因子IL-4及IFN-γ浓度降低;ALA-PDT能有效治疗CA,同时因皮疹减轻或消退可能会间接导致患者外周血NKT细胞比值、细胞因子IL-4和干扰素IFN-γ的浓度不同程度升高。     

老年慢性牙周炎合并冠心病患者龈沟液及血清中IL-1β、TNF-α、IL-18及IFN-γ的表达与临床意义

目的探讨白细胞介素(IL)-1β、肿瘤坏死因子(TNF)-α、IL-18和干扰素(IFN)-γ在老年慢性牙周炎合并冠心病患者龈沟液及血清中的表达及临床意义。方法选取45例老年慢性牙周炎合并冠心病患者(A组),45例为单纯的老年慢性中、重度牙周炎患者(B组)。另外选取同期进行健康体检的健康老年受试者45例(C组)。对三组受试者的出血指数、探诊深度进行检查比较。同时采集三组受试者的龈沟液和肘部静脉血,检测并对比三组受试者龈沟液及血清中的IL-1β、TNF-α、IL-18和IFN-γ水平。探讨牙周指数与龈沟液及血清中IL-1β、TNF-α、IL-18及IFN-γ浓度间的相关性。结果 A组、B组出血指数、探诊深度均显著高于C组,且A组显著高于B组,差异有统计学意义(P0.05)。A组、B组龈沟液、血清中的IL-1β、TNF-α、IL-18、IFN-γ水平均显著高于C组,A组显著高于B组,差异有统计学意义(P0.05)。Spearman相关性分析结果表明,出血指数、探诊深度与龈沟液、血清中的IL-1β、TNF-α、IL-18、IFN-γ均呈正相关(P0.05)。牙周指数与龈沟液、血清中的IL-1β、TNF-α、IL-18、IFN-γ均呈正相关性(P0.05)。结论老年慢性牙周炎合并冠心病患者的龈沟液、血清中的IL-1β、TNF-α、IL-18及IFN-γ均显著升高,且存在明显的相关性,提示慢性牙周炎与冠心病间可能存在一定的相关性,两者的发病可能与体内IL-1β、TNF-α、IL-18、IFN-γ等浓度升高有一定关联。     

干扰素α联合病毒唑治疗慢性丙型肝炎中Th1/Th2表达的变化

测定Th1细胞因子IFN-γ、TNF-,αTh2细胞因子IL-4、IL-10在干扰素α联合病毒唑治疗慢性丙型肝炎(CHC)中表达的变化并分析其意义。用半定量逆转录聚合酶链反应技术检测40例正常对照、48例CHC患者的外周血单个核细胞内IFN-γ、TNF-α、IL-4、IL-10 mRNA表达水平的相对含量。结果发现CHC患者Th1类细胞因子IFN-γ和TNF-α的表达水平显著低于正常对照组;治疗后完全应答组Th1类细胞因子IFN-γ和TNF-α的表达水平显著增高。提示CHC患者的细胞免疫功能低下,可能是病毒持续感染的原因之一;有效激活患者的细胞免疫功能与获得良好的治疗效果密切相关。     

Relative expression and correlation of tumor necrosis factor-α, interferon-γ, and interleukin-17 in the rheumatoid synovium

Although tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), and interleukin-17 (IL-17) play important roles in RA, their relative expression and possible correlation in synovial tissues are not well understood. In this study, mRNA expression levels of IFN-γ, IL-17, and TNF-α were investigated in individual patients with RA and the correlations between pairs of these three pro-inflammatory cytokines were analyzed. Synovial tissues were obtained during arthroplasties from 24 joints of 24 RA patients. After harvesting synovial tissues, total RNA was isolated then quantitative real-time polymerase chain reaction (qRT-PCR) for IFN-γ, IL-17, and TNF-α was performed. Correlation of expression levels between them was also analyzed. Expression levels of TNF-α, IFN-γ, and IL-17 in patients receiving TNF inhibitors (TNFi) and those treated with conventional synthetic disease-modifying antirheumatic drugs (csDMARDs) alone were also compared between groups. Based on relative expression levels of the three pro-inflammatory cytokines, patients were classified into three major types; an IFN-γ plus TNF-α-dominant type, an IL-17-dominant type, and the other type. TNF-α expression levels were correlated with IFN-γ. In addition, there was a negative correlation between TNF-α and IL-17, and IFN-γ and IL-17. Median relative expression levels of TNF-α have no significant difference between the TNFi and the csDMARDs groups. In the rheumatoid synovial tissues, expression levels of TNF-α were modulated in parallel with IFN-γ, and TNF-α and IL-17, or IFN-γ and IL-17 did not co-express at high levels. This characteristic expression pattern of the three pro-inflammatory cytokines may be clinically useful information in the current cytokine-targeted treatment with biological DMARDs for RA.     

Kinetics of phytohemaglutinin-induced IFN-γand TNF-α expression in peripheral blood mononuclear cells from patients with chronic hepatitis B after liver transplantation

AIM: To study the association between host immunity and hepatitis B virus (HBV) recurrence after liver transplantation.METHODS: Peripheral blood mononuclear cells (PBMC) were isolated from 40 patients with hepatitis B and underwent orthotopic liver transplantation (OLT) before and 2, 4, 8 wk after surgery. After being cultured in vitro for 72 h, the levels of INF-γ and TNF-α in culture supernatants were detected with ELISA. At the same time, the quantities of HBV DNA in serum and PBMCs were measured by real time PCR.RESULTS: The levels of INF-γ and TNF-α in PBMC culture supernatants decreased before and 2, 4 wk after surgery in turns (INF-γ 155.52±72.32 ng/L vs14.76±9.88 ng/L vs 13.22±10.35 ng/L, F= 6.946, P = 0.027＜0.05; TNF-α80.839±46.75 ng/L vs 18.59±17.29 ng/L vs9.758±7.96 ng/L,F= 22.61, P= 0.0001＜0.05). The levels of INF-γ and TNF-α were higher in groups with phytohemagglutinin (PHA) than in those without PHA before surgery. However,the difference disappeared following OLT. Furthermore,INF-γ and TNF-α could not be detected in most patients at wk 4 and none at wk 8 after OLT. The HBV detection rate and virus load in PBMlC before and 2, 4 wk after surgery were fluctuated (HBV detected rate: 51.4%, 13.3%, 50% respectively; HBV DNA: 3.55±0.674 log(10) copies/mL vs 3.00±0.329 log(10) copies/mL vs 4.608±1.344 log(10) copies/mL, F= 7.582, P= 0.002＜0.05). HBV DNA in serum was 4.48±1.463 log(10) copies/mL before surgery and ＜103 copies/mL after OLT except for one with 5.72×106 copies/mL 4 wk after OLT who was diagnosed as HBV recurrence.The levels of INF-γ, and TNF-α were lower in patients with a high HBV load than in those with a low HBV load (HBV DNA detected/undetected in PBMCs: IFN-γ 138.08±72.44 ng/L vs 164.24±72.07 ng/L, t = 1.065, P = 0.297＞0.05, TNF-α 80.75±47.30 ng/L vs74.10±49.70 ng/L, t= 0.407, P= 0.686＞0.05; HBV DNA positive/negative: IFN-γ 136.77±70.04 ng/L vs 175.27±71.50 ng/L, t= 1.702, P= 0.097＞0.05; TNF-α 75.37±43.02 ng/L vs 81.53±52.46 ng/L, t = 0.402,P = 0.690＞0.05).CONCLUSION: The yielding of INF-γ and TNF-α from PBMCs is inhibited significantly by immunosuppressive agents following OLT with HBV load increased, indicating that the impaired immunity of host is associated with HBV recurrence after OLT.     

Induction of HLA-DR Antigen Expression on Human Colonic Epithelium by Tumor Necrosis Factor-or and Interferon-γ

Ishii N, Chiba M, Iizuka M, Horie Y, Masamune O. Induction of HLA-DR antigen expression on human colonic epithelium by tumor necrosis factor-α and interferon-γ. Scand J Gastroenterol 1994;29:903-907.

Background: We investigated the effects of tumor necrosis factor-α (TNF-α-) and interferon-γ (IFN-γ) on HLA-DR expression on human normal colonic epithelium, using an organ culture technique. Methods: Biopsy specimens from normal colonic mucosa were cultured for 24 h with various concentrations of TNF-α and/or IFN-γ. Epithelial HLA-DR antigens were identified by an indirect immunoperoxidase staining method. Results: No expression of epithelial HLA-DR antigens was observed in specimens cultured with medium alone. Concentrations of 10³U/ml or higher of TNF-α- and 10² U/ml or higher of IFN-γ induced expression of HLA-DR antigens in a dose-dependent manner. Expression of HLA-DR was increased in specimens cultured with both TNF-α and IFN-γ compared with those cultured with either cytokine alone. Conclusions: Our findings suggest that both TNF-α and IFN-γ play important roles in regulating the expression of HLA-DR on colonic epithelium in patients with inflammatory bowel disease.     

慢性阻塞性肺疾病急性加重期合并肺结核患者外周血清中IL-6、TNF-α和IFN-γ水平变化及临床意义

目的 探究慢性阻塞性肺疾病急性加重期(AECOPD)合并肺结核患者外周血清中IL-6、肿瘤坏死因子α(TNF-α)和干扰素γ(IFN-γ)水平变化及临床意义.方法 将研究对象分为AECOPD合并肺结核组、AECOPD组、肺结核组和健康对照组,根据肺结核患者影像学检查结果将肺结核患者分为单侧病变组、双侧病变组、无空洞组和有空洞组,根据AECOPD合并肺结核患者治疗后的情况分为好转组和恶化组.采用ELISA法检测各组患者血清中的IL-6、TNF-α和IFN-γ水平.结果 AECOPD合并肺结核组血清中的IL-6和TNF-α水平显著高于AECOPD组和肺结核组(P＜0.05);单侧病变组患者血清中的IL-6、TNF-α和IFN-γ的水平显著低于双侧病变组(P＜0.05);无空洞组患者血清中的IL-6、TNF-α和IFN-γ的水平显著低于有空洞组(P＜0.05);病情好转组治疗后IL-6、TNF-α和IFN-γ水平显著低于治疗前(P＜0.05).结论 IL-6、TNF-α和IFN-γ在AECOPD合并肺结核的发病机制中起着一定的作用,其外周血清中水平的动态变化在一定程度上可以反映AECOPD合并肺结核病情的发展与转归.     

HCV感染患者免疫细胞部分细胞因子分泌情况的初步研究

目的研究与正常人比较丙型肝炎病毒（HCV）感染患者免疫细胞分泌γ干扰素（IFN-γ）、白细胞介素10（IL-10）、肿瘤坏死因子α（TNF-α）及白细胞介素4（IL-4）的细胞频数情况,了解HCV感染对其影响。方法分离外周血单核细胞（PBMCs）,应用IL-10、IFN-γ、TNF-α及IL-4流式抗体进行细胞内因子染色,应用FACSCalibur流式细胞仪及FACSCalibur软件进行检测分析。结果HCV感染患者分泌IL-10、IFN-γ、IL-4的细胞频数在CD4＋CD8-T细胞、CD4-CD8＋T细胞、NK细胞和NKT细胞均发生明显下降;分泌TNF-α的细胞频数在CD4-CD8＋T淋巴细胞及NK细胞出现下降;HCV感染患者NK细胞、NKT细胞分泌IL-10和IFN-γ的细胞频数较CD4＋CD8-T淋巴细胞、CD4-CD8＋T淋巴细胞下降得更加明显。结论HCV感染患者的细胞免疫功能受到明显的损害,细胞因子分泌能力明显减低;固有免疫细胞功能受损可能是HCV感染慢性化的重要原因;细胞因子分泌的调节是抗HCV感染免疫治疗过程中需要调节的方向。     

慢加急性乙型肝炎肝衰竭患者肝组织IFN-γ、TNF-α和IL-10的表达

目的研究乙型肝炎慢加急性肝衰竭（ACLF）患者肝内促炎细胞因子和抗炎细胞因子的表达特性。方法采用免疫组化法检测乙型肝炎ACLF患者、慢性乙型肝炎患者和正常人肝组织IFN-γ、TNF-α和IL-10的表达。结果在ACLF患者肝组织IFN-γ和TNF-α阳性细胞数均明显高于慢性乙型肝炎患者和正常对照组（P均〈0.001）;ACLF患者、慢性乙型肝炎患者和正常人肝组织IL-10的表达无统计学差异（P〉0.05）;肝内IFN-γ表达与TNF-α具有明显的相关性（r=0.886,P〈0.001）。结论乙型肝炎ACLF患者因肝内促炎细胞因子和抗炎细胞因子表达的失衡而发生了肝损伤。     

Nuclear factor-κB decoy oligodeoxynucleotides attenuates ischemia/reperfusion injury in rat liver graft

AIM: To evaluate the protective effect of NF-κB decoy oligodeoxynucleotides (ODNs) on ischemia/reperfusion (I/R) injury in rat liver graft.METHODS: Orthotopic syngeneic rat liver transplantation was performed with 3 h of cold preservation of liver graft in University of Wisconsin solution containing phosphorothioated double-stranded NF-κB decoy ODNs or scrambled ODNs. NF-κB decoy ODNs or scrambled ODNs were injected intravenously into donor and recipient rats 6 and 1 h before operation,respectively. Recipients were killed 0 to 16 h after liver graft reperfusion. NF-κB activity in the liver graft was analyzed by electrophoretic mobility shift assay (EMSA). Hepatic mRNA expression of TNF-α, IFN-γand intercellular adhesion molecule-1 (ICAM-1) were determined by semiquantitative RT-PCR. Serum levels of TNF-α and IFN-γ were measured by enzyme-linked immunosorbent assays (ELISA). Serum level of alanine transaminase (ALT) was measured using a diagnostic kit. Liver graft myeloperoxidase (MPO) content was assessed.RESULTS: NF-κB activation in liver graft was induced in a time-dependent manner, and NF-κB remained activated for 16 h after graft reperfusion. NF-κB activation in liver graft was significant at 2 to 8 h and slightly decreased at 16 h after graft reperfusion. Administration of NF-κB decoy ODNs significantly suppressed NF-κB activation as well as mRNA expression of TNF-α, IFN-γ and ICAM-1 in the liver graft. The hepatic NF-κB DNA binding activity [presented as integral optical density (IOD) value] in the NF-κB decoy ODNs treatment group rat was significantly lower than that of the I/R group rat (2.16±0.78 vs 36.78±6.35 and 3.06±0.84 vs 47.62± 8.71 for IOD value after 4 and 8 h of reperfusion, respectively, P＜0.001).The hepatic mRNA expression level of TNF-α, IFN-y and ICAM-1 [presented as percent of β-actin mRNA(%)] in the NF-κBdecoy ODNs treatment group rat was significantly lower than that of the I/R group rat (8.31 ±3.48 vs 46.37±10.65 and 7.46± 3.72 vs 74.82±12.25for hepatic TNF-α mRNA, 5.58±2.16 vs 50.46±9.35 and6.47±2.53 vs 69.72±13.41 for hepatic IFN-γ mRNA, 6.79±2.83 vs 46.23±8.74 and 5.28±2.46 vs 67.44±10.12for hepatic ICAM-1 mRNA expression after 4 and 8 h of reperfusion, respectively, P＜0.001). Administration of NF-κB decoy ODNs almost completely abolished the increase of serum level of TNF-α and IFN-γ induced by hepatic ischemia/reperfusion, the serum level (pg/mL)of TNF-α and IFN-γ in the NF-κB decoy ODNs treatment group rat was significantly lower than that of the I/R group rat (42.7±13.6 vs 176.7±15.8 and 48.4±15.1 vs216.8±17.6 for TNF-α level, 31.5±12.1 vs 102.1±14.5and 40.2±13.5 vs 118.6±16.7 for IFN-γ level after 4 and8 h of reperfusion, respectively, P＜0.001). Liver graft neutrophil recruitment indicated by MPO content and hepatocellular injury indicated by serum ALT level were significantly reduced by NF-κB decoy ODNs, the hepatic MPO content (A655) and serum ALT level (IU/L) in the NF-κB decoy ODNs treatment group rat was significantly lower than that of the I/R group rat (0.17±0.07 vs 1.12±0.25 and 0.46±0.17 vs 1.46±0.32 for hepatic MPO content, 71.7±33.2 vs 286.1±49.6 and 84.3±39.7 vs467.8±62.3 for ALT level after 4 and 8 h of reperfusion,respectively, P＜ 0.001).CONCLUSION: The data suggest that NF-κB decoy ODNs protects against I/R injury in liver graft by suppressing NF-κB activation and subsequent expression of proinflammatory mediators.     

Tumor necrosis factor-α overproduction in Fanconi's anemia

Various in vitro studies and clinical observations suggest that Fanconi's anemia (FA) patients are unable to detoxify adequately superoxide anions (O) released by activated phagocytes. Recent studies have shown that certain lymphokines such as tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) can significantly enhance O production by phagocytic cells. To ascertain lymphokine production in FA patients, we measured TNF-α and IFN-γ production in vivo and in vitro. TNF-α was detected in the plasma of 16 of 18 FA patients with concentrations ranging from 6 to 131 pg/ml (mean 31 pg/ml). TNF-α was detected in only one of 25 control (healthy donor) plasma, and the level was very low (7 pg/ml). IFN-γ levels in normal and patient plasma were negligible. Spontaneous and phytohemagglutinin (PHA)-induced production of IFN-γ and TNF-α by cultured peripheral blood mononuclear cells did not differ significantly between FA patients and normal controls. The significance of overproduction of TNF-α in vivo in the pathophysiology of FA is discussed. © 1993 Wiley-Liss, Inc.     

Detection of tumor necrosis factor-α in bone marrow plasma and peripheral blood plasma from patients with aplastic anemia

Plasma levels of interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α) were determined in healthy individuals and patients with aplastic anemia (ApAn). IFN-γ was not detected in normal peripheral blood plasma (PBP) or bone marrow plasma (BMP) and was present in PBP from only 2 of 22 patients and in BMP from 1 of 14 patients and the levels were low (< 1.5 U/ml). Elevated levels of TNF-α were present in BMP and PBP from patients but not in control (healthy donor) PBP and BMP. Eleven of twenty-four patients had elevated levels of TNF-α in their PBP and 6 of 13 patients had detectable levels of TNF-α in their BMP. Only one of the 14 healthy control donors had detectable TNF-α and the level was very low (7 pg/ml), while 13 of the 27 ApAn patients had detectable TNF-α (P = .009, chi-square test). Not surprisingly, the centers of the distributions of TNF-α concentrations of the controls and ApAn patients differed significantly (P < .017 for control and patient PBP and P < .056 for control and patient BMP, Wilcoxon rank-sum test). Spontaneous production of IFN-γ and TNF-α by cultured bone marrow mononuclear cells was observed in four of seven patients but not in the six healthy controls (P = 0.026). Spontaneous production of IFN-γ and TNF-α by cultured peripheral blood mononuclear cells from patients and controls was however similar. Phytohemagglutinin (PHA)-induced production of IFN-γ and TNF-α by cultured mononuclear cells did not differ significantly between ApAn patients and normal controls. The significance of overproduction of TNF-α in the pathophysiology of ApAn is discussed. © 1994 Wiley-Liss, Inc.