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Objective:To establish the detection method for the interferon stimulated genes(ISGs), calculate the cut-off value and test it in clinical practice.Methods:Patients with type I interferonopathies who were admitted to Peking Union Medical College Hospital from November 2017 to September 2021 were chosen as the disease group, and healthy children were included as the control group. A total of 18 children were in the disease group, including 8 males and 10 females, with a median age of 8.5 years for the first test. From them 25 blood specimens were collected. A total of 28 healthy children, aged 1 to 18 years, with a median age of 10.5 years, including 15 males and 13 females, were included in the control group. Blood samples of 34 controls and 18 interferonopathies patients were collected, then total RNA extraction and cDNA synthesis were performed. Real-time quantitative polymerase chain reaction assays were run in duplicate to measure the expression of six ISGs: interferon induced protein with tetratricopeptide repeats 1 (IFIT1), interferon α inducible protein 27 (IFI27), interferon induced protein 44 like (IFI44L), interferon stimulated genes 15 (ISG15), sialic acid binding Ig like lectin 1 (SIGLEC1), and radical S-adenosyl methionine domain containing 2 (RSAD2). The relative abundances of each target transcript was normalized to the expression level of β-Actin and OAZ. The median fold change of the six ISGs was used to create an interferon score (IS) for each individual. Samples with abnormal expressions were removed and the cDNA mix of the remaining samples was used as a calibrator to calculate the IS. We define an abnormal IS as being greater than+2 standard deviations above the mean of controls. Differences in IS between groups were compared using t-test or Mann-Whitney U-test. Results:The mean IS of controls was 1.046, standard 0.755, and the cut-off value was 2.556. A total of 25 samples from 18 interferonopathies patients were tested. The mean value was 27.010 with a 15/18 abnormality rate. Compared with the control group, IS in patients was significantly higher, t=4.247( P=0.000 1). The accuracy, precision, sensitivity, and specificity were 91.30% (42/46), 7.47%(0.084/1.124), 15/18, and 96.43% (27/28), respectively. Conclusion:This study provides a new and reliable method for clinical screening and dynamic monitoring of type Ⅰ interferonopathies by detecting ISGs expression and creating an IS.
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The clinical, imaging, genetic, therapeutic and prognostic features of a case of pediatric stroke who was finally diagnosed with Aicardi-Goutières syndrome (AGS) in Xi′an International Medical Center Hospital on October 24, 2021 were reported. A 10-year-old boy was admitted to the hospital due to weakness of the right limb for more than 10 hours. The pre-hospital CT showed multiple patchy calcifications in the bilateral frontal lobe and the right parietal lobe cortex-medullary junction. The physical examination on admission had chilblains on the hands, feet and face. National Institutes of Health Stroke Scale Score was 4 points. Brain magnetic resonance imaging showed acute brainstem infarction, no abnormality in magnetic resonance angiography, ultrasound and electrocardiogram of heart and neck vessels were normal, cerebrospinal fluid biochemistry and routine examination were normal, blood routine, biochemistry, coagulation, autoantibody series, thyroid function, tumor markers, human immunodeficiency virus and syphilis examinations were normal. After oral administration of aspirin anti-platelet aggregation and rehabilitation exercises, the muscle strength returned to normal and the patient was discharged. One month later, the result of genetic testing was reported as AGS caused by TREX1 gene mutation, and the mutation site is c.58G>A. AGS is a rare autoimmune hereditary encephalopathy with a large heterogeneity of clinical manifestations. When a hereditary disease was suspected, genetic testing should be done.
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Objective To detect mRNA expression of stimulator of interferon genes(STING)and type Ⅰ interferons(IFN-α and IFN-β)in peripheral blood mononuclear cells(PBMCs)of patients with chronic hepatitis B(CHB), and evaluate its correlation with hepatitis B virus load.Methods 88 untreated CHB patients(CHB group)and 74 healthy persons(control group)who performed physical examination were chosen from Renmin Hospital of Wuhan University during the same period between February 2016 and February 2017.Expressions of mRNA of STING, IFN-α, and IFN-βwere detected by quantitative real-time polymerase chain reaction(PCR), their relative expression values were obtained by 2-ΔΔCT method, results were statistically analyzed.Results The expression of STING, IFN-α, and IFN-βmRNA in peripheral blood of CHB patients were 2.95, 3.14, and2.01folds of healthy controls respectively, differences were statistically significant(t=-4.72, -3.41, -2.31, respectively, all P＜0.05).STING relative expression in patients with HBV DNA load≤104 IU/mL was 2.98, 3.76, and 3.97 folds of patients with HBV DNA load 104-105 IU/mL, 105-106 IU/mL, and＞106 IU/mL, respectively(P＜0.05).mRNA expressions of STING in CHB patients were positively correlated with that of IFN-αand IFN-βmRNA (r=0.475, 0.503, respectively, both P＜0.05).Conclusion The expression of STING increased in patients with CHB, high expression of STING impacted the replication of HBV.
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Objective To detect mRNA expressions of STING and type Ⅰ interferons(IFN-α and IFN-β) in peripheral blood mononuclear ceils (PBMCs) of patients with chronic hepatitis C(CHC).Methods 113 patients with CHC and 94 healthy controls were collected from Renmin Hospital of Wuhan University during January 2015 and January 2016.Expressions of mRNA of STING,RIG-1,IFN-α and IFN-β were detected by real-time quantitative PCR,and their relative expression values were obtained by 2-△△Ct method and the results were statistically analyzed.Results The expression of mRNA of STING,RIG-1,IFN-α and IFN-β in CHC were 0.018,0.361,0.578 and 0.573 times than its in healthy controls respectively and the differences were statistically significant (t-8.28,4.26,2.18 and 2.07,all P ＜ 0.05).Pearson correlation analysis showed that mRNA expressions of STING in healthy controls were positively correlated with that of IFN-α mRNA and IFN-β mRNA (r=0.487 and 0.207,all P＜0.05),which had no correlation in CHC (r=0.174 and 0.091,all P＞0.05).The expressions of STING mRNA was positively correlated with that of RIG-1 mRNA in healthycontrols (r=0.222,P＜0.05),but there was no correlation of expression in CHC between STING mRNA and RIG-1 mRNA (r=-0.029,P＞0.05).Conclusion Compared with healthy controls,the expression of STING,RIG-1,IFN-α and IFN-β mRNA were all reduced.There was positive correlation between STING and RIG-1,IFN-α and IFN-β in healthy controls,but no correlation in CHC.
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Objective To detect the expression of retinoic acid-inducible gene Ⅰ ( RIG-Ⅰ) in peripheral blood mononuclear cells (PBMCs) of patients with chronic hepatitis C (CHC), and to explore its correlations with type Ⅰinterferon gene expression and serum level of hepatitis C virus .Methods A total of 101 na?ve patients with CHC were enrolled from Renmin Hospital of Wuhan University during July 2014 and July 2015, and 69 healthy subjects served as controls .The expressions of RIG-Ⅰ, IFNαand IFNβmRNA in PBMCs and serum level of HCV RNA were detected by real-time quantitative polymerase chain reaction.Pearson correlation analysis was performed to investigate the correlations of RIG-Ⅰ mRNA expression with IFNα/βmRNA expression and serum level of HCV RNA .Results The relative expressions of RIG-Ⅰ, IFNα, IFNβmRNA to healthy controls in CHC group were 0.082, 0.022 and 0.156, respectively (t=7.83, 3.65 and 2.13, all P0.05).Conclusion Expression of RIG-ⅠmRNA decreases in patients with CHC , and is positively correlated with that of IFNαand IFNβmRNA.
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At present, the etiology and pathogenesis of multiple sclerosis are unclear. RIG-Ⅰ-like receptors are a new-ly discovered pattern recognition receptors (RLRs), which are located in cytoplasm. They can recognize the helicase of viral dsRNAs, and interact with interferon beta promoter stimulator (IPS)-1 through their caspase activation recruitment domain (CARD), then form IPS-1 signalsome and induce the expression of interferon typeⅠ(Ⅰ-IFN), thereby initiate innate im-mune response and induce antiviral response. Recent studies have found that mice lacking IPS-1 would develop exacerbated disease and accompanied by markedly higher inflammation, increasing axonal damage and demyelination. Furthermore, initi-ating the RIG-Ⅰ-like helicase receptor on the immune cells can alleviate inflammation and myelin fracture in multiple scle-rosis of mouse model, thus limit the incidence of paralysis. This paper is a review about the research progress on RLRs in the treatment of multiple sclerosis.
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Objective To explore whether different Epstein-Barr virus (EBV) infection status is involved in systemic lupus erythematosus (SLE) pathogenesis through type Ⅰ interferon pathway by observing the EBV antibodies,serum interferon α (IFN-α) level and four type Ⅰ interferon induced gene (ISGs;2'-5' oligoadenylate synthetase-like protein (OASL),myxovirous resistant 1 (MX1),interferon-stimulated gene 15 (ISG15) and lymphocyte antigen 6 complex,locus E (LY6E) expressions in SLE patients and healthy controls.Methods Forty-eight patients with SLE and 36 healthy controls were enrolled into this study.The serum antibodies of EBV capsid antigen-IgG/lgM and EBV nuclear antigen 1-IgG,and serum levels of IFN-α were measured by enzyme linked immunosorbent assay (ELISA) method.Real time reverse transcription polymerase chain reaction was used to test the mRNA levels of OASL,MX1,ISG15 and LY6E in the peripheral blood lymphocytes (2-△△Ct was used to indicate the gene expression).Statistical analysis was performed using t-test or Mann-Whitney U test,Chi square test and Spearman correlation test.Results ① The EBV lytic infection rate (VCA-IgM) in SLE patients was higher than that in healthy controls (40％ vs 11％,x2=5.381,P=0.027).② The serum levels of IFN-α in SLE patients were higher than those in the healthy controls [(206±151) ng/L vs (90± 76) ng/L,t=4.248,P＜0.05],as well as the mRNA levels of OASL,MX1,ISG15 and LY6E [1.8(0.6,5.1) vs 1.2 (0.5,1.4);1.9(1.0,4.4) vs 0.9(0.7,2.5);4.1(1.6,7.8) vs 0.8(0.5,1.7);1.6(0.7,3.3) vs 0.8(0.6,1.2),U=604,560,312,608;P＜0.05,respectively].The mRNA levels of OASL,MX1,LY6E and ISG15 were all positively related to SLE disease activity index (SLEDAI) scores (r=0.319,0.461,0.547,0.484,P＜0.05,respectively).Serum IFN-α levels were elevated in SLE patients with EBV lytic infection than in non-lytic infection patients [(282± 174) ng/L vs (157±114) ng/L,t=2.604,P＜0.05];The mRNA levels of OASL and ISG15 were also elevated in patients with lytic EBV infection [2.0(0.8,7.6) vs 1.2(0.6,3.1);6.2(2.4,15.5) vs 3.3(1.3,6.3),U=377,350,385,354;P＜0.05,respectively].The SLEDAI scores in patients with EBV lyric infection were higher than in patients with non-lyric infection (16±4 vs 12±8,P＜0.05).Conclusion EBV infection may be involved in the pathogenesis of SLE by activating the type Ⅰ interferon pathway.
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Objective To explore the expression of interferon-inducible genes (IFIG) in idiopathic inflammatory myopathies (IIM) patients and its clinical significance.Methods Peripheral blood mononuclear cells from 52 IIM patients and 20 healthy controls were extracted.The transcriptional levels of six IFIG genes:oligoadenylate synthetase 1 (OAS-1),IFN-induced protein with tetratricopeptide repeats 1(IFIT1),IFIT4,interferon-inducible-protein 44 (IFI44),radical S-adenosyl methionine domain containing protein 2 (RSAD2) and myxo-virus resistance gene 1 (MX-1) were detected by real-time polymerase chain reaction (PCR).Interferon scores were calculated and compared.The correlations between interferon scores and clinical features of IIM patients were investigated.Non-parametric Mann-Whitney and Spearman analysis were used for statistical analysis.Results In IIM patients,the transcriptional levels of OAS-1 [90.4(40.2,350.8)],IFIT1 [112.7(49.8,151.1)],IFIT4 [127.8(110.4,200.2)],IFI44 [100.3(39.5,160.7)],RSAD2 [100.9(50.2,559.2)] and MX-1 [110.7(60.3,298.9)] were greatly higher than those in healthy controls [4.5(2.1,27.8),21.5(0.4,31.4)i 21.5(0.8,51.2),5.2(0.4,27.8),27.8(17.5,33.5),25.1(15.7,31.8),respectively] (Z=-5.121,-4.359,-5.301,-4.906,-4.348,-4.151; P＜0.01).Meanwhile,interferon scores were significantly elevated in IIM patients [65.7(3.6,136.9),Z=-5.778,P＜0.01],compared with healthy controls [1.2(0.1,3.1)],especially in those with interstitial lung disease [89.3(41.9,176.7),Z=0.544,P＜0.05].Interferon scores were correlated positively with ESR (r=0.234,P＜0.05).Conclusion The mRNA levels of OAS-1,IFIT1,IFIT4,IFI44,RSAD2 and MX-1 are significantly increased in patients with IIM.Interferon score might be used as a new biomarker for IIM patients.
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Objective To investigate mRNA expressions of Toll-like receptors (TLR3 and TLR7)and type Ⅰ interferons (IFN-α and IFN-β) in peripheral blood mononuclear cells (PBMCs) of patients with chronic hepatitis C (CHC).Methods Thirty patients with CHC and 30 healthy controls were collected from Renmin Hospital of Wuhan University during September 2013 and May 2014.Liver function was detected using enzyme-linked immunosorbent assay (ELISA).mRNA expressions of TLR3,TLR7,IFN-α and IFN-β were detected by real-time quantitative PCR,and their relative expression values were obtained by 2-△△Ctmethod.The t test was performed for the comparison of mean differences between CHC patients and healthy controls,and Pearson analysis was used to analyze the correlations between TLR mRNA and IFN mRNA,liver function,HCV RNA load.Results Taking mRNA expressions in healthy control group as 1,the relative mRNA expressions of TLR3,TLR7,IFN-α and IFN-β in CHC group were 0.086,0.633,0.145 and 0.423 respectively (t =4.25,2.39,2.37 and 2.80,all P ＜ 0.01).Pearson correlation analysis showed that mRNA expressions of TLR3 and TLR7 were positively correlated with that of IFN-β (r =0.381 and 0.487,all P ＜ 0.05),but not correlated with IFN-α mRNA expression,HCV RNA load,ALT and AST (all P ＞ 0.05).Conclusion The mRNA expressions of TLR3,TLR7 and IFN-α,IFN-β decrease in CHC patients,and the mRNA expressions of TLR3 and TLR7 are positively correlated with that of IFN-β,indicating that increasing the expression of TLRs might be a new strategy in CHC treatment.
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ObjectiveTo investigate the expression levels of the type Ⅰ IFN system in muscle and lung of experimental autoimmune myositis (EAM) model and to evaluate whether the type Ⅰ IFN system associates with the pathogenesis of the EAM model in rats.MethodsThe EAM model was established to determine creatine kinase (CK) in blood serum.The pathology of muscle and lung tissue was examined by hematoxylin-eosin staining.The concentration of type Ⅰ IFN system mRNA in muscle and lung tissue was detected by real-time PCR.ResultsThe concentration of CK in model group [(209.17 ±91.95) IU/L]was significantly higher than that of two control groups(P ＜0.05).The scores of muscle and lung in EAM model were significantly higher than that of control groups (all P ＜ 0.05).The expression levels of the type Ⅰ IFN system in muscle of EAM model were significantly higher than that of control groups(all P ＜0.05).The expression levels of the type Ⅰ IFN system in muscle with EAM model were positively correlated with CK and the scores of muscle (all P ＜ 0.05).The expression levels of IFNα, IFNβ, IFNαR1, signal transducer and activator of transcription 1 (STAT1), myxovirus resistance protein 1 (MX1) in lung of EAM model were significantly higher than those of control groups(P ＜ 0.05), but not seen in INF-induced protein with tetratricopetide repeats 1 (IFIT1) and IFN-stimulated gene 15 (ISG15).The expression levels of IFNα, IFNβ, IFNαR1, STAT1 and MX1 in lung with EAM model were positively correlated with the scores of lung pathology (all P ＜ 0.05).ConclusionThe type Ⅰ IFN system probably played a crucial role in the pathogenesis and the pathology of muscle and lung of EAM modeL
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Objective To study 6 type Ⅰ interferon (IFN)-inducible genes (IFIT4, IFI44, Ly6e,OAS1, OAS2 and OAS3) in patients with lupus nephritis (LN) and analyze its correlated expression levels with disease activity and/or clinical manifestations. Methods Total RNA was obtained simultaneously from kidney tissues and peripheral blood cells of 12 patients with diffuse proliferative lupus nephritis and 10 normal controls. Moreover, peripheral blood cells were obtained from 119 LN patients and 35 normal controls. Total RNA was extracted and reversely transcribed into complementary DNA. Gene expression levels were measured by real-time polymerase chain reaction by comparing to a housekeeping gene, and IFN score was calculated. Disease activity was determined by the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI). Results The 6 genes were highly expressed in diffuse proliferative lupus nephritis patients compared with normal controls. IFN scores were positively correlated with SLEDAI score, the concurrent presences of anti-double-stranded DNA (anti-dsDNA) antibodies (P<0.05) and hypocomplementemia (P<0.01). Conclusion The 6 IFN-inducible genes are highly expressed iri LN patients. IFN scores are elevated in active lupus nephritis patients, in patients with positive anti-ds-DNA antibody and hypocomplementemia. IFN scores may be a useful biomarker for lupus nephritis therapy.
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Objective To detect the expression of type Ⅰ interferon in monocyte-derived dendritic cells(MoDCs)after Toll like receptor(TLR)3 triggered in patients with chronic hepatitis B(CHB),and to evaluate immune responses of CHB patients and its roles in the mechanisms of persistent infection of hepatitis B virus(HBV)and chronicity of hepatitis.Methods Peripheral blood mononuclear cells(PBMCs)were isolated and purified using magnetic beads(plasma was saved simultaneously)from 26 CHB patients and 18 healthy volunteers(HV).Dendritic cells(DCs)were induced and proliferated in a culture medium with recombinant human granulocyte macrophage colony stimulating factor(rhGM-CSF)and recombinant human interleukin(rhIL-4).EX3s were stimulated with Poly Ⅰ:C and the supernatants were collected at 0 h and 24 h after stimulation.Type Ⅰ interferon(IFN-α and IFN-β)in plasma and supernatants were examined by enzyme linked immunosorbent assay (ELISA).Results The levels of type Ⅰ interferon in plasma were not significantly different in groups of HV and CH B.IFN-α and IFN-β expressions in supernatants before Poly Ⅰ:C stimulation were(80.00±16.15)ng/L,(36.39±13.90)ng/L in CHB group and(76.76±15.90)ng/L,(37.14±13.68)ng/L in HV group,respectively.And there were no statistical differences between two groups(t=1.651,t=0.178;both P＞0.05).IFN-α expressions in supernatants at 24 h after stimulation in two groups were both higher than those before stimulation(at 0 h),but there were no statistical differences(t=1.534,t=1.243;both P＞0.05).IFN-β expressions in supernatants at 24 h after stimulation in HV group was(54.57±16.80)ng/L,which was significantly higher than that at 0 h(37.14±13.68)ng/L(t=4.061,P＜0.05).However,there was no significant difference at 24 h than tht at 0 h in CHB group(t=1.796,P＞0.05).At 24 h after stimulation.IFN-β level was(54.57±16.80)ng/L in HV group,which was significantly higher than that[(41.64±12.57)ng/L]in CHB group(t=2.921,P＜0.05).Conclusions Functions of MoDCs from CHB patients are impaired and MoDCs could not express type Ⅰ interferon normally.Expression of type Ⅰ interferon after TLR3 triggered in CHB patients is mainly IFN-β.
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Objective To observe the clinical efficacy and safety of topical interferon alfa-1b cream in the treatment of herpes zoster. Methods A randomized, double-blind, parallel placebo-controlled clinical study was conducted. The test drug was topically used in herpes zoster patients, three times a day for 2 weeks. Patients whose skin lesions cleared completely were followed for 29 days to observe postherpetic neuralgia. Results One hundred and twenty-eight patients with herpes zoster were enrolled into this trial. Sixty-five patients were randomly selected to receive interferon alfa-1b cream and sixty-three patients received vehicle cream. After following up for 11, 14, 22, 29 days the cure rates were 69.2%, 81.5%, 90.8%, 95.4% respectively in the study group and were 57.1%, 71.4%, 84.1% and 84.1% respectively in the control group(P
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Toll-like-receptors(TLRs) have been found to play a critical role in immune response of hosts.They can trigger innate immune responses by detecting pathogen-associated molecular patterns(PAMPs).This paper reviews the roles of several toll-like receptors(TLR4,TLR5,and TLR9) in radioprotection and the related mechanisms,so as to provide theoretical evidences for further studying their roles in radioprotection.