Evaluation of Interferon-Gamma Release Assay Testing and Tuberculin Skin Test for Early Diagnosis of Tuberculosis in Children and Adolescents

Background: This study aimed to evaluate the diagnostic value of interferon-γ release assay (IGRA), a sensitive microbiological diagnostic method, in children and adolescents with suspected tuberculosis in a country with a high burden of tuberculosis. Method: This study included 581 children and adolescents aged 4 - 19 years who were suspected of having tuberculosis, were latently infected with Mycobacterium tuberculosis, and had received at least one dose of BCG vaccine between April 17, 2019, and February 24, 2021. The study evaluated the TST results of 106 patients who had a positive Quantiferon test and were suspected of having tuberculosis. Results: The study included 581 patients aged between 4 and 19 years. Of these, 106 patients tested positive for the Quantiferon test, while 19 were indeterminate and 456 were negative. The Quantiferon test positivity rate was 18.24%. Among the 106 QFT-Plus-positive cases, 23 patients also tested positive for TST. The difference in distribution was found to be statistically significant. Conclusion: The QFT-Plus test is considered an alternative to TST and other microbiological diagnostic methods for early tuberculosis diagnosis, particularly in children and adolescents.

慢加急性乙型肝炎肝衰竭患者细菌感染病原菌分布及血清PCT、IFN-γ和IL-6水平变化分析

目的调查慢加急性乙型肝炎肝衰竭(HBV-ACLF)患者细菌感染病原菌分布特征,分析血清降钙素原(PCT)、干扰素-γ(IFN-γ)和白细胞介素-6(IL-6)水平预测细菌感染的效能。方法2019年12月~2023年1月我院收治的HBV-ACLF患者86例,常规分离和鉴定菌种,采用电化学发光免疫法检测血清PCT水平,采用ELISA法检测血清IFN-γ和IL-6水平,应用受试者工作特征(ROC)曲线评估血清指标联合预测HBV-ACLF患者发生细菌感染的效能。结果本组发生细菌感染者37例,未发生明确感染者49例;在37例HBV-ACLF并发细菌感染患者中,共检出68株感染病原菌,其中革兰氏阴性菌41株(60.3%),革兰氏阳性菌27株(39.7%);感染组血清PCT、IFN-γ和IL-6水平分别为(10.9±3.1)μg/L、(46.5±1.9)pg/mL和(16.9±1.6)pg/mL,均显著高于未感染组【分别为(0.9±0.1)μg/L、(20.1±2.4)pg/mL和(4.8±0.9)pg/mL,P<0.05】,28 d和90 d病死率分别为67.6%和75.7%,均显著高于未感染组(分别为8.2%和12.2%,P<0.05);经ROC分析显示,分别以PCT>3.3μg/L、IFN-γ>45.5 pg/mL和IL-6>15.4pg/mL为截断点,其联合预测HBV-ACLF患者并发细菌感染的曲线下面积(AUC)为0.874,诊断的灵敏度为93.6%,特异度为84.1%。结论HBV-ACLF患者细菌感染的病原菌分布具有一定的特征性,以革兰氏阴性菌感染为主,除检测血清PCT水平外,监测血清IFN-γ和IL-6水平可能有助于早期发现细菌感染而给予预防性处理,或能提供生存率。

藏獒IFN-γ基因在毕赤酵母中的表达与抗病毒活性分析

采用毕赤酵母表达系统表达藏獒γ-干扰素(Tibetan mastiff interferon gamma,TmIFN-γ)并分析其生物活性,为抗病毒生物制品的研发与临床应用奠定基础。将构建的TmIFN-γ的真核表达质粒pPIC9k-TmIFN-γ转化至毕赤酵母GS115细胞,用不同浓度的G418筛选得到多拷贝重组菌株;利用甲醇进行诱导表达,以镍层析柱纯化目的蛋白,对表达产物进行SDS-PAGE和Western blotting鉴定;通过CCK-8试验检测重组TmIFN-γ蛋白的细胞毒性作用,利用VSV-MDBK系统检测其抗病毒生物活性。结果显示,成功筛选得到高拷贝pPIC9k-TmIFN-γ重组转化毕赤酵母菌株,利用甲醇诱导可获得TmIFN-γ重组蛋白,该蛋白对细胞无毒性,具有显著抗病毒活性,其抗病毒生物效价为1.727×10~5IU/mL。本研究获得了具有抗病毒活性的TmIFN-γ,有助于进一步探索TmIFN-γ的功能和开发新型基因工程抗病毒药物。

Clinical and bacteriological correlates of whole blood interferon gamma(IFN-γ) in newly detected cases of pulmonary TB

Objective:To determine the relationship of the capacity to produce interferon gamma(IFN-γ) in whole blood,bacteriological,hematological,radiographic and clinical presentations in new,HIV seronegative cases of pulmonary tuberculosis(TB).Methods:80 cases and 50 control subjects aged 15 years onwards,representative of Kasturba Hospital and Nursing schools of Wardha district of Maharashtra state in India were examined for their health condition with standard methodology.Results:Among these TB patients,73.8%were Quantiferon-TB gold (QFT) positive with IFN-γconcentration as 0.35 IU or more and there was none in healthy controls.The mean IFN-γconcentrations varied between 9.58 IU(50-59 yrs) and 2.58 IU(≥60 yrs),showing no trend.The differences in positivity and mean IFN-γconcentrations were statistically insignificant.Both the QFT positivity and IFN-γconcentrations were higher in normal lymphocyte percent as compared to below and above normal,but differences were not statistically significant.Conclusions:The IFN-γconcentrations are not correlated with any of the predictors of disease severity studied,the levels are significantly higher in observation group as compared to healthy group.

血清IL-17A、IFN-γ与慢性鼻窦炎伴鼻息肉严重程度的相关性

目的探讨血清白细胞介素-17A(IL-17A)、γ-干扰素(IFN-γ)与慢性鼻窦炎伴鼻息肉(CRSwNP)患者病情严重程度的相关性。方法以河南科技大学第一附属医院2021年6月至2023年6月收治的82例CRSwNP患者为观察组,另择同期收治的30例单纯鼻中隔偏曲的患者为对照组。于治疗前测定并对比两组血清IL-17A、IFN-γ、转化生长因子-β1(TGF-β1)、基质金属蛋白酶-9(MMP-9)水平;并采用pearson相关性分析CRSwNP患者血清IL-17A、IFN-γ与病情严重程度及血清TGF-β1、MMP-9的关系,采用受试者工作特征(ROC)曲线分析血清IL-17A、IFN-γ对CRSwNP的诊断效能。结果观察组的血清IL-17A、IFN-γ、TGF-β1、MMP-9均明显高于对照组,差异有统计学意义(P<0.05);观察组的Lund-Mackay评分、Lund-Kennedy评分均明显高于对照组,差异有统计学意义(P<0.05)。Pearson相关性分析显示,CRSwNP患者的血清IL-17A、IFN-γ水平与Lund-Mackay评分、Lund-Kennedy评分、血清TGF-β1、MMP-9水平均呈正相关关系(P<0.05);ROC曲线分析显示,血清IL-17A、IFN-γ水平及二者联合检测均对CRSwNP有诊断效能(P<0.05),且相较于单项检测,二者联合检测的效能更高,AUC为0.953,敏感度为93.90%,特异度为90.00%。结论CRSwNP的发生伴随IL-17A、IFN-γ升高,IL-17A、IFN-γ水平与病情严重程度及鼻黏膜组织重构有关,还可作为CRSwNP的早期诊断指标。

产褥期感染危险因素分析及TAC、IFN-γ及α1-AG预测价值

目的:分析产褥期感染危险因素并探讨总抗氧化能力(TAC)、干扰素γ(IFN-γ)及α1酸性糖蛋白(α1-AG)预测价值。方法:回顾性收集2020年6月-2023年1月本院接诊的250例产妇临床资料,根据产褥期感染发生情况分为感染组(n=31)和未感染组(n=219),收集临床资料,分析产褥期感染危险因素,检测血清TAC、IFN-γ、α1-AG水平并采用受试者工作特征曲线(ROC)分析其预测产褥期感染价值。结果:单因素分析,两组年龄、孕周、产次、内分泌疾病比较无差异(P>0.05);两组产程、妇科炎症、妊娠高血压疾病、妊娠期糖尿病、产后出血、居住地、分娩方式、胎膜早破、卧床时间比较有差异(P<0.05);感染组血清IFN-γ(150.05±36.25 pg/ml)、α1-AG(156.35±40.91 mg/dl)高于未感染组(134.71±30.56 pg/ml、116.25±38.36 mg/dl),TAC(8.20±1.50 kU/ml)低于未感染组(10.83±2.45 kU/ml)(均P<0.05);白细胞计数、C反应蛋白及降钙素原水平产前两组无差异(P>0.05),产后两组均升高且感染组高于未感染组(P<0.05)。多因素非条件logistic分析,产程长、妇科炎症、妊娠高血压疾病、妊娠期糖尿病、产后出血、居住地为乡村、分娩方式为剖宫产、胎膜早破、卧床时间≥3d、TAC降低、IFN-γ升高及α1-AG升高均是产褥期感染发生的独立危险因素(P<0.05);ROC结果显示,预测产褥期感染血清TAC的曲线下面积(AUC)为0.808,灵敏度76.3%,特异度79.5,截断值9.56 kU/ml;血清IFN-γ的AUC为0.651,灵敏度61.5%,特异度69.4%,截断值141.24 pg/ml;血清α1-AG的AUC为0.753,灵敏度73.4%,特异度70.8%,截断值130.35 mg/dl。结论:产程长、妇科炎症、妊娠高血压疾病、妊娠期糖尿病、产后出血、居住地为乡村、剖宫产、胎膜早破、卧床时间长均是产褥期感染的独立危险因素,血清TAC、IFN-γ、α1-AG在产褥期感染者血清中表达异常且可对产褥期感染的发生有一定预测价值。

清眩润目饮对干眼模型鼠眼表炎性因子IFN-γ、IL-17、MMP3、MMP9的影响

目的 对雌性C57BL/6J小鼠进行干眼造模,给予中药清眩润目饮治疗,观察其对眼表炎性因子干扰素-γ(IFN-γ)、白细胞介素-17(IL-17)、基质金属蛋白酶3(MMP3)、基质金属蛋白酶9(MMP9)的影响。方法 将健康雌性C57BL/6J小鼠随机分为A组(空白对照组)、B组(模型组)、C组(中药组)、D组(玻璃酸钠组),每组20只。B、C、D组进行干眼造模,C组给予18 mg/(kg·d)的中药灌胃处理,每天2次;D组给予0.1%玻璃酸钠滴眼液,每天2次。实验结束后取小鼠的泪腺、角膜,采用免疫组化法检测IFN-γ、IL-17、MMP3、MMP9水平。结果 在小鼠泪腺组织中,与B组比较,C组IFN-γ及IL-17蛋白表达水平降低(P <0.05),MMP3蛋白表达水平显著降低(P <0.01);与B组比较,D组IFN-γ蛋白表达水平显著降低(P <0.01),MMP9蛋白表达水平降低(P <0.05)。在小鼠角膜组织中,在小鼠角膜组织中,与B组比较,C组MMP3蛋白表达水平降低(P <0.05),MMP9蛋白表达水平显著降低(P <0.01);与B组比较,D组MMP9蛋白表达水平显著降低(P <0.01)。结论 清眩润目饮能够有效下调干眼模型鼠眼表炎性因子IFN-γ、IL-17、MMP3、MMP9水平,减轻干眼眼表损伤。

重组鼠IFN-γ腺病毒诱发细胞因子释放综合征小鼠模型的建立和观察

目的:通过向C57Bl/6J小鼠腹腔注射IFN-γ腺病毒(Ad-mIFN-γ)建立细胞因子释放综合征(CRS)的动物模型。方法:构建Ad-mIFN-γ及对照Ad-lacZ腺病毒载体,分别以MOI=100体外转染小鼠腹腔巨噬细胞,流式细胞术检测其对细胞mIFN-γ分泌的影响。将40只雌性C57Bl/6J小鼠按随机数字表法分为对照组、载体对照组、病毒低、中、高剂量组(每组8只),分别向各组小鼠腹腔注射PBS(200μL)、Ad-lacZ(2×10^(7)PFU/只)、Ad-mIFN-γ(5×10^(6)PFU/只)、Ad-mIFN-γ(1.5×10^(7)PFU/只)和Ad-mIFN-γ(2×10^(7)PFU/只)。每日观测小鼠的体质量及生存情况;第3天时采用流式细胞术检测小鼠外周血和脾内单核细胞(CD11b^(+))、巨噬细胞(CD11b^(+)/CD86^(+))比例,免疫荧光染色法检测脾内CD11b^(+)的单核细胞比例;第9天时采用流式细胞术检测小鼠血清中细胞因子的分泌水平;第14天,采用颈椎脱臼法处死小鼠,H-E染色法观察小鼠肝、脾、肺和肾的病理和组织学变化。结果:Ad-mIFN-γ体外感染小鼠腹腔巨噬细胞,在第3天检测到巨噬细胞分泌mIFN-γ达到峰值(118.34±2.90)pg/mL,并在一周内持续高分泌mIFN-γ,Ad-lacZ对照组IFN-γ分泌水平较低后,第3天时为(0.17±0.08)pg/mL。小鼠腹腔注射Ad-mIFN-γ后,在14 d内病毒低、中剂量组无小鼠死亡,病毒高剂量组小鼠体质量持续减轻(P<0.001);第3天,病毒高剂量组小鼠外周血和脾组织内单核细胞、巨噬细胞比例较对照组和中剂量组均显著增加(P<0.05或P<0.01);第9天,病毒低、中、高剂量组小鼠血清中mIFN-γ、IL-6、单核细胞趋化蛋白-1(MCP-1)、IL-1、TNF-α等细胞因子的水平均显著升高(P<0.001);10 d内病毒高剂量组小鼠死亡率达100%。组织病理检测可见病毒高剂量组小鼠的肝、脾、肺、肾组织有明显损伤。结论:Ad-mIFN-γ体外感染小鼠原代腹腔巨噬细胞后,可以快速分泌mIFN-γ;腹腔注射高剂量(2×10^(7)PFU/只)Ad-mIFN-γ导致小鼠出现CRS典型表现,可作为CAR-T细胞治疗诱发CRS的动物模型。

ITGα3和IFN-γ与非小细胞肺癌患者临床病理特征及预后的关系

目的分析整合素α3(ITGα3)和血清γ干扰素(IFN-γ)与非小细胞肺癌患者临床病理特征及预后的关系。方法选取2020年10月—2022年10月本院收治的非小细胞肺癌患者86例作为观察组,另选取于接受检查的良性病变患者36例为对照组。比较不同分组和不同预后患者血清ITGα3和血清IFN-γ水平并分析其水平阳性、阴性与患者临床病理特征的关系。Cox比例风险回归模型分析ITGα3和血清IFN-γ水平对患者生存预后的影响。结果观察组患者IFN-γ阳性表达率为61.63%,显著高于对照组的13.89%(χ^(2)=23.190,P<0.05)。观察组患者ITGα3阳性表达率为55.81%,显著高于对照组的11.11%(χ^(2)=20.736,P<0.05)。IFN-γ的阳性表达与性别、年龄、分化程度无相关性(P>0.05),与吸烟史、TNM分期、病理分型、淋巴结转移存在显著相关性(P<0.05)。ITGα3的阳性表达与性别、年龄、TNM分期、分化程度无相关性(P>0.05),与吸烟史、病理分型、淋巴结转移存在显著相关性(P<0.05)。死亡组患者IFN-γ和ITGα3表达显著高于非死亡组(P<0.05)。Cox回归模型分析显示,IFN-γ、ITGα3阳性表达为影响NSCLC患者生存预后的独立危险因素(P<0.05)。结论ITGA3和血清IFN-γ与吸烟史、病理分型、淋巴结转移存在显著相关性,IFN-γ、ITGα3阳性表达为影响NSCLC患者生存预后的独立危险因素,通过观察其水平有利于病情判断,对临床评估治疗效果有一定帮助。

IL-1β、IL-6、IFN-γ在甲型H1N1流感病毒性肺炎患儿血清中的表达及其与疾病预后的关系研究

目的 研究白细胞介素(IL)-1β、IL-6、γ干扰素(IFN-γ)在甲型H1N1流感病毒性肺炎患儿血清中的表达及其与疾病预后的关系。方法 选取40例甲型H1N1流感病毒性肺炎患儿作为观察组,另选取体检健康儿童40例作为对照组。所有研究对象均行酶联免疫吸附试验(ELISA)法检测IL-1β、IL-6、IFN-γ水平。比较两组IL-1β、IL-6、IFN-γ表达水平;比较IL-1β、IL-6、IFN-γ单一检测与联合检测对甲型H1N1流感病毒性肺炎的诊断效能;比较观察组不同预后(痊愈、有效、无效)患儿IL-1β、IL-6、IFN-γ表达水平。结果 观察组IL-1β、IL-6、IFN-γ表达水平分别为(32.56±3.72)pg/ml、(15.80±3.36)pg/ml、(12.15±3.13)μg/L,均高于对照组的(6.39±1.05)pg/ml、(4.35±0.50)pg/ml、(3.61±0.42)μg/L(P<0.05)。IL-1β对甲型H1N1流感病毒性肺炎的诊断灵敏度为80.00%,特异度为77.50%;IL-6对甲型H1N1流感病毒性肺炎的诊断灵敏度为75.00%,特异度为77.50%;IFN-γ对甲型H1N1流感病毒性肺炎的诊断灵敏度为77.50%,特异度为80.00%;联合检测对甲型H1N1流感病毒性肺炎的诊断灵敏度为95.00%,特异度为55.00%;联合检测对甲型H1N1流感病毒性肺炎的诊断灵敏度均高于IL-1β、IL-6、IFN-γ单一检测,特异度低于IL-1β、IL-6、IFN-γ单一检测(P<0.05)。观察组患儿治疗后痊愈18例,有效17例,无效5例。其中痊愈患儿IL-1β、IL-6、IFN-γ表达水平均低于有效及无效患儿,有效患儿IL-1β、IL-6、IFN-γ表达水平均低于无效患儿(P<0.05)。结论 IL-1β、IL-6、IFN-γ与甲型H1N1流感病毒性肺炎患儿病情有关,上述指标对判断患儿预后有着重要价值。

弥漫大B细胞淋巴瘤患者血浆IFN-γ、IL-2及外周血T淋巴细胞亚群、NK细胞与其临床分期的关系

目的探讨弥漫大B细胞淋巴瘤(DLBCL)患者血浆干扰素γ(IFN-γ)、白细胞介素-2(IL-2)水平及外周血T淋巴细胞亚群、自然杀伤(NK)细胞与临床分期的关系。方法纳入河南中医药大学第三附属医院2020年12月至2022年12月收治的DLBCL患者198例,根据临床分期分成Ⅰ期组(31例)、Ⅱ期组(56例)、Ⅲ期组(59例)、Ⅳ期组(52例)。比较4组血浆IFN-γ、IL-2及外周血CD3^(+)、CD3^(+)CD4^(+)T、CD3^(+)CD8^(+)T、NK细胞水平,分析各指标与DLBCL分期的相关性。结果Ⅲ期组、Ⅳ期组的血浆IFN-γ、IL-2水平低于Ⅰ期组、Ⅱ期组(P<0.05)。Ⅲ期组、Ⅳ期组外周血CD3^(+)、CD3^(+)CD4^(+)T、CD3^(+)CD8^(+)T、NK细胞水平低于Ⅰ期组、Ⅱ期组(P<0.05)。DLBCL患者IFN-γ、IL-2与CD3^(+)、CD3^(+)CD4^(+)T、CD3^(+)CD8^(+)T、NK细胞水平呈正相关(P<0.05)。DLBCL患者的血浆IFN-γ、IL-2及外周血CD3^(+)、CD3^(+)CD4^(+)T、CD3^(+)CD8^(+)T、NK细胞水平与临床分期呈负相关(P<0.05)。IFN-γ、IL-2、CD3^(+)、CD3^(+)CD4^(+)T、CD3^(+)CD8^(+)T、NK细胞水平增高是控制临床分期增高的保护因素(P<0.05)。结论DLBCL患者的临床分期与血浆IFN-γ、IL-2及外周血T淋巴细胞亚群、NK细胞水平相关,且血浆IFN-γ、IL-2水平与T淋巴细胞亚群、NK细胞存在相关性,上述指标有望对DLBCL进展风险进行预测。

Assessment by meta-analysis of interferon-gamma for the diagnosis of tuberculous peritonitis

AIM:To investigate the performance and diagnostic accuracy of interferon-gamma(IFN-γ) for tuberculous peritonitis(TBP) by meta-analysis.METHODS:A systematic search of English language studies was performed.We searched the following electronic databases:MEDLINE,EMBASE,Web of Science,BIOSIS,LILACS and the Cochrane Library.The Standards for Reporting Diagnostic Accuracy initiative and Quality Assessment for Studies of Diagnostic Accuracy tool were used to assess the methodological quality of the studies.Sensitivity,specificity,and other measures of the accuracy of IFN-γ concentration in the diagnosis of peritoneal effusion were pooled using random-effects models.Receiver operating characteristic(ROC) curves were applied to summarize overall test performance.Two reviewers independently judged study eligibility while screening the citations.RESULTS:Six studies met the inclusion criteria.The average inter-rater agreement between the two reviewers for items in the quality checklist was 0.92.Analysis of IFN-γ level for TBP diagnosis yielded a summary estimate:sensitivity,0.93(95%CI,0.87-0.97);specificity,0.99(95%CI,0.97-1.00);positive likelihood ratio(PLR),41.49(95%CI,18.80-91.55);negative likelihood ratio(NLR),0.11(95%CI,0.06-0.19);and diagnostic odds ratio(DOR),678.02(95%CI,209.91-2190.09).χ 2 values of the sensitivity,specificity,PLR,NLR and DOR were 5.66(P = 0.3407),6.37(P = 0.2715),1.38(P = 0.9265),5.46(P = 0.3621) and 1.42(P = 0.9220),respectively.The summary receiver ROC curve was positioned near the desirable upper left corner and the maximum joint sensitivity and specificity was 0.97.The area under the curve was 0.99.The evaluation of publication bias was not significant(P = 0.922).CONCLUSION:IFN-γ may be a sensitive and specific marker for the accurate diagnosis of TBP.The level of IFN-γ may contribute to the accurate differentiation of tuberculosis(TB) ascites from non-TB ascites.

Effectiveness of interferon-gamma release assays for differentiating intestinal tuberculosis from Crohn's disease:A meta-analysis

AIM:To investigate the clinical usefulness of interferon-gamma release assays(IGRAs)in the differential diagnosis of intestinal tuberculosis(ITB)from Crohn’s disease(CD)by meta-analysis.METHODS:A systematic search of English language studies was performed.We searched the following databases:Medline,Embase,Web of Science and the Cochrane Library.The Standards for Reporting Diagnostic Accuracy initiative and Quality Assessment for Studies of Diagnostic Accuracy tool were used to assess the methodological quality of the studies.Sensitivity,specificity,and other measures of the accuracy of IGRAs in the differential diagnosis of ITB from CD were pooled and analyzed using random-effects models.Receiver operating characteristic curves were applied to summarize overall test performance.Two reviewers independently judged study eligibility while screening the citations.RESULTS:Five studies met the inclusion criteria.The average inter-rater agreement between the two reviewers for items in the quality checklist was 0.95.Analysis of IGRAs for the differential diagnosis of ITB from CD produced summary estimates as follows:sensitivity,0.74(95%CI:0.68-0.80);specificity,0.87(95%CI:0.82-0.90);positive likelihood ratio,5.98(95%CI:3.79-9.43);negative likelihood ratio,0.28(95%CI:0.18-0.43);and diagnostic odds ratio,26.21(95%CI:14.15-48.57).The area under the curve was 0.92.The evaluation of publication bias was not significant(P=0.235).CONCLUSION:Although IGRAs are not sensitive enough,they provide good specificity for the accurate diagnosis of ITB,which may be helpful in the differential diagnosis of ITB from CD.

Animal experiment and clinical study of effect of gamma-interferon on hepatic fibrosis

AIM: To evaluate the antifibrotic effect of different doses of recombinant human Gamma-Interferon (IFN-gamma) in two rat models of hepatic fibrosis, and to observe its effect on moderate chronic hepatitis B virus fibrosis. METHODS: Hepatic fibrosis was successfully induced in 150 and 196 rats by subcutaneous injection of carbon tetrachloride (CCl4) and intraperitoneal injection of dimethylnitrosamine (DMN), respectively. Each of the two model groups was divided into: (1) fibrotic model group; (2) colchicine treatment group (0.1 mg/kg/day, gastrogavage for 8 weeks); (3) high-dose IFN-gamma group (15 MU/kg per day, i.m. for 8 weeks); (4) medium-dose IFN-gamma group (5 MU/kg daily, i.m. for 8 weeks); and (5) Y low-dose IFN-gamma group (1.67 MU/kg daily, i.m. for 8 weeks). Another group of 10 rats without any treatment was used as normal controls. At the end of the experiment, semi-quantitative histopathological scores of inflammation and fibrosis, liver alpha smooth muscle actin (alpha-SMA) expression level, liver hydroxyl proline content and serum hyaluronic acid levels were compared. And 47 medium chronic hepatitis B viral fibrosis patients were studied. They were given IFN-gamma treatment, 100 MU/day i.m. for the first three months and 100 MU qod i.m. for the next six months. Semi-quantitative pathological scores of inflammation and fibrosis and serum hepatic fibrosis indices were compared within the 9 months. RESULTS: In animal experiment, the pathological fibrosis scores and liver hydroxyl proline content were found to be significantly lower in rats treated with different doses of IFN-gamma as compared with rats in fibrotic model group induced by either CCl4 or DMN, in a dose-dependent manner. For CCl4-induced model, pathological fibrosis scores in high, medium and low doses IFN-gamma groups were 5.10 +/- 2.88, 7.70 +/- 3.53 and 8.00 +/- 3.30, respectively, but the score was 14.60 +/- 7.82 in fibrotic model group. Hydroxyl proline contents were 2.83 +/- 1.18, 3.59 +/- 1.22 and 4.80 +/- 1.62, in the three IFN-gamma groups, and 10.01 +/- 3.23 in fibrotic model group. The difference was statistically significant (P【0.01). Similar results were found in DMN-induced model. Pathological fibrosis scores were 6.30 +/- 0.48, 8.10 +/- 2.72 and 8.30 +/- 2.58, in high, medium and low doses IFN-gamma groups, and 12.60 +/- 3.57 in fibrotic model group. Hydroxyl proline contents were 2.72 +/- 0.58, 3.14 +/- 0.71 and 3.62 +/- 1.02, in the three IFN-gamma groups, and 12.79 +/- 1.54 in fibrotic model group. The difference was statistically significant (P【0.01).Serum hepatic fibrosis indices decreased significantly in the 47 patients after IFN-gamma treatment (HA: 433.38 +/- 373.00 vs 281.57 +/- 220.48; LN: 161.22 +/- 41.02 vs 146 +/- 35 +/- 44. 67; PC III: 192.59 +/- 89.95 vs 156.98 +/- 49.22; C-I: 156.30 +/- 44.01 vs 139.14 +/- 34.47) and the differences between the four indices were significant (P 【0.05). Thirty-three patients received two liver biopsies, one before and one after IFN-gamma treatment. In thirty of 33 patients IFN-gamma had better effects according to semi-quantitative pathological scores (8.40 +/- 5.83 vs 5.30 +/- 4.05, P【0.05). CONCLUSION: All the three doses of IFN-gamma are effective in treating rat liver fibrosis induced by either CCl4 or DMN, the higher the dose, the better the effect. And IFN-gamma is effective for patients with moderate chronic hepatitis B viral fibrosis.

Performance and correlation of interferon gamma release assays and tuberculin skin test in HIV-infected children and adolescents with immune reconstitution

Objective:To evaluate the performance of interferon gamma release assays and tuberculin skin test in HIV-infected children and adolescents with immune reconstitution.Methods:A cross-sectional study was conducted in HIV-infected patients aged 5-18 years receiving antiretroviral treatment with CD4 T-lymphocytes>25%or>500 cells/mm3 for at least 6 months.QuantiF ERON-TB Gold,T-SPOT.TB,and tuberculin skin test were performed in each patient.Results:A total of 50 patients were enrolled with median age of 13.7 years,CD4 counts of 753(IQR:587-989)cells/mm3.Among 27 patients with tuberculosis(16)or tuberculosis exposure(11),8(29.6%)were positive to at least one test,2(7.4%)were positive QuantiFERON-TB Gold,3(11.1%)positive T-SPOT.TB,and 7(25.9%)had tuberculin skin test≥5 mm.Among 23 patients without history of tuberculosis or exposure,all had negative interferon gamma release assays,while 2(8.7%)had positive tuberculin skin test.Conclusions:All tests had low sensitivity despite immune reconstitution.

Effect of recombinant human growth hormone and interferon gamma on hepatic collagen synthesis and proliferation of hepatic stellate cells in cirrhotic rats

BACKGROUND: Fibrosis plays a key role in the development of liver cirrhosis. In this study, we investigated the effect of growth hormone and interferon gamma on hepatic collagen synthesis and the proliferation of hepatic stellate cells in a cirrhotic rat model. METHODS: Cirrhosis was induced in rats using carbon tetrachloride. Rats were simultaneously treated with daily subcutaneous injections of recombinant human growth hormone or interferon gamma combined with recombinant human growth hormone. The control group was given saline. The relative content of type I and type IV collagen was assessed by indirect immunofluorescence analysis. Activated hepatic stellate cells were prepared from cirrhotic rats. The 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide (MTT) method was used to assess the effects of recombinant human growth hormone and interferon gamma on these cells in vitro. RESULTS: Both qualitative and quantitative analysis showed that type I and type IV collagen secretion increased with time after recombinant human growth hormone administration and was significantly higher than control and recombinant human growth hormone combined with interferon gamma administration. In vitro, recombinant human growth hormone significantly stimulated hepatic stellate cell proliferation in a concentration-dependent manner (10 -3 -10 -1 mg/100 μL), andinterferon gamma (10 -2 -10 -1 μg/100 μL) significantly inhibited their growth compared to the control group. Interferon gamma combined with recombinant human growth hormone eliminated this growth-promoting effect to a certain degree in a concentration-dependent manner (10 -1 μg/100 μL, P<0.05, 10 -2 -10 -3 μg/100 μL, P>0.05) and a time-dependent manner (P<0.05). CONCLUSIONS: Recombinant human growth hormone increased collagen secretion in cirrhotic rats in vivo and promoted the proliferation of hepatic stellate cells from cirrhotic rats in vitro. It is possible that concurrent interferon gamma therapy can offset these side-effects of recombinant human growth hormone.

The Expression of Interleukin-17, Interferon-gamma, and Macrophage Inflammatory Protein-3 Alpha mRNA in Patients with Psoriasis Vulgaris

Summary: To investigate the role of Interleukin-17 (IL-17), Interferon-gamma (IFN-γ), and macrophage inflammatory protein-3 alpha (MIP-3α) in the pathogenesis of psoriasis, reverse transcriptase-polymerase chain reaction (RT-PCR) was used to semi-quantitatively analyze the mRNA expression of IL-17, IFN-γ, and MIP-3α in 31 psoriatic lesions and 16 normal skin tissues. The results showed that the mRNA of the three cytokines was present in all specimens. And the expression level of IL-17 mRNA in skin lesions was 1.1416±0.0591, which was significantly higher than that in normal controls (0.8788±0.0344, P<0.001). The expression levels of IFN-γ mRNA were 1.1142±0.0561 and 0.9050±0.0263, respectively, with significant difference(P<0.001). And the expression levels of MIP-3α mRNA in psoriatic lesions was 1.1397±0.0521, which was markedly higher than that in normal controls (0.8681±0.0308, P<0.001). These findings indicate that up-regulated expression of IL-17, IFN-γ, and MIP-3α might be involved in the pathogenesis of psoriasis.

Retinal stem cells transplantation combined with copolymer-1 immunization reduces interferon-gamma levels in an experimental model of glaucoma

AIM: To explore the effect of immunization with copolymer-1 (COP-1) and retinal stem cells (RSCs) transplantation on interferon-gamma (IFN-gamma) levels in a rat experimental glaucoma model. METHODS: An experimental glaucoma was induced by argon laser photocoagulation of the episcleral veins and limbal plexus in the right eye of rats. Immediately following glaucoma induction, rats were immunized with COP-1. RSCs were cultured and transplanted intravitreally into the eyes of glaucoma model animals 1 week post-laser treatment. Six experimental groups were used: COP-1/RSC, PBS/RSC, COP-1/PBS, PBS/PBS, glaucoma model group, and a normal control group. The concentration of IFN-gamma in aqueous humor (AH) and serum was measured by enzyme-linked immunosorbent assay (ELISA) in each of the six groups. Retinal ganglion cell (RGC) survival was assessed by quantifying apoptosis using Hoechst staining. RESULTS: Concentrations of IFN-gamma in AH and serum of rats that had undergone glaucoma induction were higher than those of non-induced control rats. The concentrations of IFN-gamma in AH and serum of the COP-1/RSCs treated group were determined to be 2371.9ng/L and 710.9ng/L, respectively, which were significantly lower than those in the other treated groups (P<0.05). In fact, IFN-gamma levels in the dual treated group were reduced to background levels. The COP-1/RSC group had lower number of apoptotic RGCs than the other three experimental groups (P<0.05). CONCLUSION: The reduced levels of IFN-gamma in AH and serum of the COP-1/RSC group may be related to synergistic effects between RSCs transplantation and COP-1 immune modulation. It is likely that the lower levels of IFN-gamma prevented RGCs glaucomatous apoptasis.

Capillary Electrophoretic Immunoassay with Laser-induced Fluorescence Detection for Interferon-gamma

Capillary electrophoretic immunoassay with laser-induced fluorescence detection for recombinant human interferon-gamma (IFN-g) was established. The limits of detection for three forms of IFN-g are 6.9 ng/L, 5.7 ng/L and 5.0 ng/L, respectively.

Inhibitory effect of gold nanoparticles conjugated with interferon gamma and methionine on breast cancer cell line

Objective: To develop a gold nanoparticles complex conjugated with interferon-gamma(IFN-g) and methionine along with application of hyperthermia using near-infrared laser beams for the treatment of cancer cells.Methods: Gold nanorods(10 nm) were conjugated with IFN-g and methionine using carbodiimide family and characterized after purification by dialysis bags. Breast cancer cells were cultured and incubated with gold nanorods at different concentrations followed by irradiation with near-infrared laser beam. Samples were then evaluated for their viability in order to determine the effect of treatment and variables by MTT assy.Results: Zetasizer results confirmed the conjugation of gold nanorods with methionine and IFN-g. The median percentage of cell viability in 0.30 mg/m L concentration of gold nanorods was 82%. The cell viability reached to 85% at the same concentration of gold nanorods, which existed in the assayed complex. The results of MTT assay showed that the 0.60 mg/m L concentration of gold nanoparticles complex was toxic on tumor cells(P < 0.05). After exposure to hyperthermia, the viability of cells at 6 min decreased to77% in 0.30 mg/m L concentration of gold nanorods complex.Conclusions: The size and concentration of gold nanorods was not cytotoxic. However,their presence during irradiation near-infrared laser increased the number of dead cells during the treatment of cells.