Aberrant Activation of JAK/STAT Pathway Components in Response to G-CSF,Interferon-a/p and Interferon-y in NFS-60 Cells

Abstract

There is evidence that the cellular responses to cytokines, such as granulocyte colony stimulating factor (G-CSF) and interferons, depend on prior activation of components of the JAK/STAT signalling pathway. We report here that the myeloid cell line NFS-60 shows aberrant JAK/STAT signalling yet elicits expected biological responses to G-CSF and interferons-α/β and γ. Instead of increased phosphorylation of JAKl and JAK2 in response to G-CSF and interf eron-y, and JAK1 and JAk2 in response to interferon-α/β we observed only an increase of phosphorylation of Tyk2 in response to all of these cytokines in NFS-60 cells. The subset of STAT proteins being activated in response to these cytokines was unusual as well. G-CSF activated STAT3 and STAT5A, whereas interferons activated, in addition to STAT1 and STAT5 other, as yet unidentified, DNA binding proteins. However, NFS-60 cells show normal biological responses to these cytokines, such as proliferation in response to G-CSF, and reduction of proliferation, induction of an anti-viral response and induction of specific genes in response to interferons.     

Crucial Role of CD4+CD 25+ FOXP3+ T Regulatory Cell,Interferon-γ and Interleukin-16 in Malignant and Tuberculous Pleural Effusions

The differential diagnosis between malignant and tuberculous exudative pleural effusions is an important clinical problem. The aim of current study is to evaluate the frequencies of T-regulatory cells (Treg) on the basis of distinct phenotypes in the differential diagnosis between malignant and tuberculous pleural effusion. In addition, to evaluate Interferon-gamma (IFN- γ) and interleukin-16 (IL-16) levels and their correlation to Treg cells in malignant and tuberculous pleural effusions. Sixty patients with pleural effusion (26 tuberculous and 34 malignant) and 20 healthy controls were included in the study. Pleural fluid and peripheral blood were assessed for frequencies of T regs, IL-16, and IFN-γ. Pleural effusions from both tuberculous and malignant groups represented significantly higher levels (more in TB) for the following cell populations than peripheral blood: total lymphocytes, CD3+lymphocyte, CD4+CD25+lymphocyte and Treg (CD4+ CD25+FoxP3+). Levels of IL-16 and IFN-γ in tuberculous group were significantly higher than that in malignant group. Regulatory T cells, INF-γ and IL-16 are new important tools for differentiation between tuberculous and malignant pleural effusion.     

Type-I Interferon assessment in 45 minutes using the FilmArray® PCR platform in SARS-CoV-2 and other viral infections

Low concentrations of type-I interferon (IFN) in blood seem to be associated with more severe forms of Coronavirus disease 2019 (COVID-19). However, following the type-I interferon response (IR) in early stage disease is a major challenge. We evaluated detection of a molecular interferon signature on a FilmArray® system, which includes PCR assays for four interferon stimulated genes. We analyzed three types of patient populations: (i) children admitted to a pediatric emergency unit for fever and suspected infection, (ii) ICU-admitted patients with severe COVID-19, and (iii) healthcare workers with mild COVID-19. The results were compared to the reference tools, that is, molecular signature assessed with Nanostring® and IFN-α2 quantification by SIMOA® (Single MOlecule Array). A strong correlation was observed between the IR measured by the FilmArray®, Nanostring®, and SIMOA® platforms (r-Spearman 0.996 and 0.838, respectively). The FilmArray® panel could be used in the COVID-19 pandemic to evaluate the IR in 45-min with 2 min hand-on-time at hospitalization and to monitor the IR in future clinical trials.     

聚乙二醇干扰素alpha-2a联合恩替卡韦治疗高病毒载量HBeAg阳性慢性乙型肝炎的临床研究

目的 观察聚乙二醇干扰素alpha-2a联合恩替卡韦治疗高病毒载量HBeAg阳性慢性乙型肝炎的疗效及安全性.方法 60例HBeAg阳性患者随机分成三组:聚乙二醇干扰素α-2a组20例(A组),恩替卡韦组20例(B组),聚乙二醇干扰素联合恩替卡韦组20例(C组)(联合治疗12周后单用聚乙二醇干扰素治疗).观察三组患者治疗4、12及24周时丙氨酸氨基转移酶(ALT)复常率、HBV DNA阴转率、HBsAg和HBeAg血清转换情况,并观察其不良反应.结果 分别在治疗第4、12、24周时比较三组,A组、B组的HBV DNA阴转率均明显低于C组(分别为:Z=-4.6,P＜0.0001;Z=-2.53,P=0.0114);A组ALT复常率明显低于C组(Z=-2.63,P=0.0086),B组与C组的ALT复常率差异无统计学意义;C组的HBsAg、HBeAg下降幅度大于A组和B组.结论 聚乙二醇干扰素alpha-2a联合恩替卡韦治疗高病毒载量HBeAg阳性慢性乙型肝炎患者,6个月疗程中HBV DNA阴转率、ALT复常率均优于聚乙二醇干扰素单药治疗,且安全性良好.     

基因1b型丙型肝炎病毒NS5A区变异及其与聚乙二醇干扰素α-2a联合利巴韦林治疗应答的相关性

目的探讨基因1b型慢性丙型肝炎患者NS5A区基因变异与聚乙二醇干扰素α-2a（PEG—IFNα-2a）联合利巴韦林（RBV）抗病毒治疗疗效的关系。方法回顾性选择15例应用PEG—IFNα-2a／RBV抗病毒治疗的基因1b型慢性丙型肝炎患者,其中7例为快速应答（RVR）,8例为无应答（NR）,应用RT—PCR法扩增NS5A全长片段,用克隆测序方法进行核苷酸和氨基酸序列测定。结果没有发现与治疗应答相关的单个氨基酸或基序变异,RVR组和NR组在NS5A、ISDR、PKRBD和IRRDR区的氨基酸突变数目差别均无统计学意义,RVR组在V3区（aa2356～2379）的氨基酸突变数目高于NR组（分别为5．3±0.5和3．4±0．6,P=0．03）。结论V3区的氨基酸突变数目与PEG—IFNα-2a联合RBV抗病毒疗效可能相关。     

丙种球蛋白及干扰素治疗手足口病并发病毒性脑炎疗效观察

目的观察丙种球蛋白及干扰素在治疗小儿手足口病(HFMD)并发病毒性脑炎的临床疗效、症状改善状况及安全性。方法选取我院2012-12—2013-12收治的手足口病并发病毒性脑炎患儿87例为研究对象,随机分为3组,丙种球蛋白组30例,连续5d静滴人血丙种球蛋白,剂量1g/(kg·d);干扰素组27例,采用肌内注射重组人干扰素80μg/(次·d),持续3d;联合组30例,采取丙种球蛋白和干扰素联合治疗。3组患儿在降颅压、退热、预防细菌感染、营养脑细胞等治疗和护理方式上均相同,观察3组患儿1周内临床症状改善情况,对比临床疗效及安全性。结果 3组在生理指标恢复时间上差异无统计学意义(P0.05),在热程、疱疹消失时间和神经系统病理征转阴时间上差异有统计学意义,联用组所需时间显著少于干扰素组和丙种球蛋白组(P0.01),干扰素组和丙种球蛋白组间无显著性差异(P0.05);联用组有效率(93.33%)大于干扰素组(77.78%)和丙种球蛋白组(76.67%),但两两比较差异无统计学意义(P0.05);联用组不良反应发生率高于干扰素组和丙种球蛋白组,但3组间差异无统计学意义(P0.05)。结论小儿手足口病并发病毒性脑炎采用丙种球蛋白联合干扰素治疗的效果较单独使用明显,缩短了病程,且不良反应可耐受,具有一定临床意义。     

辅助性T淋巴细胞1、2、17在慢性乙型肝炎病毒感染状态中的变化

目的 了解慢性HBV感染中辅助性T淋巴细胞(Th)1、Th2、Thl7产生的主要细胞因子的表达,以反映这3种细胞的功能状态.方法 ELISA测定34例慢性乙型肝炎、31例肝硬化、26例原发性肝癌、22例慢性重型乙型肝炎患者及15例健康对照者m清中的1FN-γ、IL-4和IL-17水平,对数据进行t检验和Pearson相关分析.结果 IFN-γ/IL-4在慢性乙型肝炎患者(1.08±0.66)比健康对照者(2.60±0.60)降低(P＜0.01),在慢性重型乙型肝炎患者(4.81±0.87)比健康对照者升高(P＜0.01);IFN-γ/IL-17在慢性乙型肝炎(1.13±0.85,P＜0.01)、肝硬化(1.69±0.92,P=0.010)、肝癌(1.76±0.84,P=0.011)患者比健康对照者(2.66±0.70)降低,在慢性重型乙型肝炎患者(3.68±0.42)比健康对照者升高(P=0.004);IL-4/IL-17在肝硬化(0.72±0.38,P=0.026)、肝癌(0.63±0.19,P＜0.01)患者比健康对照者(1.04±0.23)降低.结论 慢性乙型肝炎患者IL-4的表达占优势;肝硬化、肝癌患者IL-17的表达占优势;慢性重型乙型肝炎患者IFN-γ的表达占优势.     

The effect of interferon on the liver in chronic hepatitis C: a quantitative evaluation of histology by meta-analysis

Background/Aims: Several randomized clinical trials of interferon in chronic hepatitis C have examined the histological changes in paired biopsy specimens. We have attempted a quantitative evaluation by meta-analysis.Methods: Randomized Clinical Trials found by MEDLINE search were included if: a) they compared different IFN regimens with non-active treatment or with each other, b) they obtained biopsies before starting and at the time of stopping IFN in a sizable proportion of the treated and control patients, and c) they assessed the biopsy-specimens semi-quantitatively according to Scheuer's numerical scoring, system or Knodell's Histological Activity Index, with quantitation of fibrosis and Iobular, portal and periportal necroinflammation.Results: Seventeen trials were identified, in which 1223 adult patients had been studied. All trials homogeneously pointed towards a favorable interferon effect. The pooled data show a statistically significant histological improvement in treated patients as compared with controls for each of the four Histological Activity Index components and for the total Histological Activity Index score (overall improvement was −0.82 in favor of interferon, p<0.0001, 95% Confidence Interval −1.25 to −0.40). In the ten trials reporting histological changes separately in biochemical responders (primary and sustained responders) and non-responders, histological improvement was confined to the subset of biochemical responders. No change or very little change occurred in non-responders.Conclusions: Interferon treatment in chronic hepatitis C significantly improves liver histology. The effect of interferon is closely related to biochemical response. Studies assessing histological outcome 1 year or more after interferon treatment in long-term responders and comparatively in non-responders or relapsers would be important to confirm the regression of the necroinflammatory process in the former, as suggested by the normal serum alanine aminotransferase levels.     

Treatment responses in Asians and Caucasians with chronic hepatitis C infection

AIM：To conduct a multicentre retrospective review of virological response rates in Asians infected with genotype 1 chronic hepatitis C（CHC） treated with combination interferon and ribavirin and then to compare their responses to that among Caucasians.
METHODS：Asian patients infected with genotype 1 CHC treated at 4 Australian centres between 2001 to 2005 were identified through hospital databases.Baseline demographic characteristics,biochemical,virological and histological data and details of treatment were collected.Sustained virological responses（SVR） in this cohort were then compared to that in Caucasian subjects,matched by genotype,age,gender and the stage of hepatic fibrosis.
RESULTS：A total of 108 Asians with genotype 1 CHC were identified.The end of treatment response（ETR） for the cohort was 79% while the SVR was 67%.Due to the relatively advanced age of the Asian cohort,only sixty-four subjects could be matched with Caucasians.The ETR among matched Asians and Caucasians was 81% and 56% respectively（P=0.003）,while the SVR rates were 73% and 36%（P 〈0.001） respectively.This difference remained significant after adjusting for other predictive variables.
CONCLUSION： Genotype 1 CHC in Asian subjects is associated with higher rates of virological response compared to that in Caucasians.