Interferon-gamma-inducible Protein 10 Levels Research Articles

IP-10, MIP1α, IL-6, and IL-1β as Biomarkers Associated with Disease Severity of COVID-19

Background: The factors responsible for the progression of COVID-19 from a mild illness to a severe and often lethal condition, characterized by respiratory failure and multiple organ involvement, remain unclear. The identification of biomarkers capable of predicting disease progression is of the highest importance. Objectives: This study sought to assess laboratory measurements of interferon-gamma inducible protein-10 (IP-10), macrophage inflammatory protein 1-alpha (MIP1α), interleukin-6 (IL-6), and interleukin-1 beta (IL-1β) to achieve this objective. Methods: We measured IP-10 and MIP1α by qRT-PCR and IL-6 and IL-1β using an enzyme-linked immunosorbent assay in 120 serum samples. We analyzed differences between patients with moderate, severe, and recovered COVID-19. Results: The number of positive cases for biomarkers IP-10, MIP1α, IL-6, and IL-1β were significantly different between groups. The expression levels of IP-10 and MIP1α were significantly higher in patients with severe COVID-19 compared to those who had recovered. A strong positive association was observed between IP-10 and MIP1α in severe infection cases. Additionally, these biomarkers were relatively independent predictors of disease severity. Conclusions: The results suggest that IP-10, MIP1α, IL-6, and IL-1β are promising research candidates for understanding the severity of COVID-19 and for investigating possible pathophysiological mechanisms of the disease.

IP-10 for the Diagnosis and Treatment Monitoring of Tuberculosis in Children.

To determine the utility of interferon-gamma-inducible protein 10 (IP-10) for identifying active tuberculosis (TB) and TB infection (TBI) in children in BCG-vaccinated populations, establish its diagnostic performance characteristics, and evaluate changes in IP-10 level during anti-TB chemotherapy. Concentrations of IP-10 and IFN-γ were measured in QuantiFERON-TB Gold (QFT) supernatants in children with suspected TB or due to recent TB contact. A total of 225 children were investigated: 33 with active TB, 48 with TBI, 83 TB contacts, 20 with suspected TB but other final diagnoses, and 41 controls. In 60 children, cytokine responses were evaluated at a follow-up visit after 2 months of anti-TB treatment. IP-10 expression was significantly higher in infected children (active TB and TBI cases) than in uninfected individuals. IP-10 proved effective in identifying TB infection at its optimal cut-off (>1084.5 pg/mL) but was incapable of differentiating between children with active TB and TBI. Combining IP-10 and IFN-γ increased the QFT sensitivity. IP-10 but not IFN-γ decreased significantly during anti-TB treatment in children with active TB (p = 0.003). IP-10 identifies TB infection and declines during anti-TB chemotherapy in children. Incorporating IP-10 into new immunodiagnostic assays could improve TB diagnosis and allow for treatment monitoring.

Profile and clinical significance of interferon gamma-inducible protein-10 (IP-10) and its receptor in patients with hepatocellular carcinoma.

Chemokines play a vital role in tumor progression, metastasis and prognosis; however, the profile and clinical significance of gamma interferon-inducible protein-10 (IP-10) and its receptor (CXCR3) in patients with hepatocellular carcinoma (HCC) have not been well evaluated. Liquid-phase chip technology was used to detect the serum IP-10 in 85 patients with HBV-related HCC, 50 patients with chronic hepatitis B (CHB) and 50 liver cirrhosis subjects (CS); simultaneously, the CXCR3 and Alpha fetoprotein (AFP) were determined. Additionally, their mRNA or protein expression levels in peripheral blood mononuclear cells (PBMC), liver tumor and paracancerous tissues were quantified using qRT-PCR or ELISA. Moreover, the IP-10 and CXCR3 expression was verified by the online data from Gene Expression Omnibus. Furthermore, the relationships of serum IP-10, CXCR3 and AFP levels with their overall survival rate were also analyzed. The levels of IP-10 and CXCR3 in HCC group were significantly higher than those in CHB and CS groups, and their mRNA of PBMC is significantly positive correlation with those in their liver tissues or HBV DNA load (P < 0.0001), respectively. The serum IP-10 and CXCR3 in HCC were significantly correlated with tumor differentiation, metastases staging and distant metastasis (P < 0.05), but not related to gender, age and tumor size (P > 0.05, except IP-10 based on age). The serum IP-10 (142.6 pg/mL) and CXCR3 (241.2 pg/mL) could be differential diagnostic surrogates that distinguish HCC from CS, and the lower IP-10 level may be conducive to the postoperative survival of HCC patients. Moreover, the IP-10 and CXCR3 would be related to anti-tumor immunity in HCC patients and be a potential target for treatment of HCC.

An ultrasensitive and long-lasting chemiluminescence immunoassay for IP-10 detection based on a 4-bromophenol-reinforced bienzymatic system

The 10-kDa chemokine interferon-gamma-inducible protein 10 (IP-10) is considered one of the most promising biomarkers for diagnosing both tuberculosis and COVID-19 infections. The blood samples of patients at different disease states contain different levels of IP-10, which need to be detected in a rapid, specific and ultrasensitive manner. Here, we report a bienzymatic chemiluminescence sandwich immunoassay (BCSI) assay for the ultrasensitive and stable detection of IP-10. In this assay, IP-10 is first efficiently captured using a double-antibody sandwich strategy. The detection antibody is linked to catalase (CAT) via a streptavidin–biotin signal amplification system to achieve highly efficient conversion of hydrogen peroxide (H2O2) to oxygen and water. In the chemiluminescence (CL) reaction, horseradish peroxidase (HRP) acts as an efficient catalyst, and 4-bromophenol acts as an enhancer for the cyclic transition of HRP, which results in a strong and durable CL signal. The bienzymatic catalysis with CAT and HRP and the potentiation of 4-bromophenol enables the assay to be ultrasensitive and stable. The CL intensity was found to be well correlated with the detection of IP-10 at levels in the range of 0.71 to 125,000 pg/mL, which covers more than 6 orders of magnitude, with a detection limit of 0.63 pg/mL. The coefficient of variation was 1.49%, and the recovery range of IP-10 in serum was 86.21%-104.57%. This assay provides a wide linear range and high sensitivity and may be a promising method for the high-throughput detection of IP-10 in the diagnosis of tuberculosis and COVID-19.

Effects of ranibizumab on growth factors and mediators of inflammation in the aqueous humor of patients with diabetic macular edema.

The study aims to investigate changes in the aqueous humor levels of 8 growth factors and inflammatory mediators after intravitreal ranibizumab injection (IRI) and the relationship between these substances and functional-morphological parameters in patients with diabetic macular edema (DME). We recruited 25 patients with DME who were scheduled to receive 2 doses of IRI at monthly intervals. At baseline and 1 month after IRI, we measured aqueous levels of vascular endothelial growth factor (VEGF), placental growth factor (PlGF), monocyte chemoattractant protein 1 (MCP-1), soluble intercellular adhesion molecule-1 (sICAM-1), platelet-derived growth factor (PDGF)-AA, interleukin (IL)-6, IL-8, and interferon-gamma inducible protein 10 (IP-10) by the suspension array method. Central macular edema (CMT) or macular volume (MV) was examined by optical coherence tomography before and 1 month after IRI, and the improvement of macular edema was evaluated by calculating the percent change of CMT or MV. Aqueous humor levels of VEGF, PlGF, PDGF-AA, and IP-10 were significantly decreased 1 month after IRI (P < 0.001, P = 0.002, P = 0.002, and P = 0.005, respectively). In addition, the baseline aqueous humor levels of PlGF, MCP-1, and IL-6 were significantly correlated with the improvement in best corrected visual acuity (P = 0.036, P = 0.024, and P = 0.049, respectively). The baseline aqueous humor level of sICAM-1 was significantly negatively correlated with the change in CMT (P = 0.005), and the baseline aqueous humor levels of VEGF and PlGF were significantly correlated with the change in MV (P = 0.020 and P = 0.003, respectively). Furthermore, the percentage reduction in VEGF after IRI was significantly correlated with the change in MV (P = 0.037). Our findings suggest that the change in aqueous humor levels of VEGF, PlGF, and ICAM-1 in DME may not only be an anatomic response but also a potential therapeutic target. This study was registered in the University Hospital Medical Information Network (UMIN) clinical trial registry. The registration number is UMIN000030301.

Effervescent Glutamine Formulation Improves the Beneficial Effects of Antiretroviral Therapies on Immune Function in HIV/AIDS Carrier Patients.

The impact of oral supplementation with an effervescent glutamine formulation on the beneficial effects of antiretroviral therapies was evaluated in people living with HIV/AIDS. For this purpose, 12 HIV/AIDS carrier patients with CD4+ T cell counts <500, and who had received the same antiretroviral therapy for at least 1 year before starting this investigation were selected. The patients were required to dissolve the effervescent glutamine formulation (supplied in sachets) in water immediately before oral ingestion (12.4 g), once a day, after lunch or after dinner during 30 days. CD4+ T cell counts, complete blood cell counts, serum cytokines, and amino acids levels were quantified; biochemical and toxicological measurements were performed. The numbers of CD4+ T cells were increased (P < .05), and the serum C-reactive protein levels decreased (P < .01) after the administration of effervescent glutamine formulation. Serum levels of interferon-gamma inducible protein-10, RANTES, and macrophage inflammatory protein-1β were decreased after the treatment with effervescent glutamine formulation. No changes were observed in the serum levels of amino acids, hematological, toxicological, and biochemical parameters. In conclusion, the treatment during 30 days with effervescent glutamine formulation was well tolerated, promoted reduction of inflammation, and improved the beneficial effects of antiretroviral therapies in HIV/AIDS carrier patients.

Plasma levels of CRP, neopterin and IP-10 in HIV-infected individuals with and without pulmonary tuberculosis

IntroductionTuberculosis (TB) is a major cause of morbidity and death worldwide, and disproportionally affects people with HIV. Many cases still remain undiagnosed, and rapid and effective screening strategies are needed to control the TB epidemics. Immunological biomarkers may contribute. MethodsPlasma samples from healthy individuals (n: 12) and from HIV-infected individuals with (n: 21) and without pulmonary TB (n: 122) were tested for C-reactive protein (CRP), neopterin, and interferon-gamma-inducible protein-10 (IP-10). Increased levels of biomarkers and WHO 4-symptom-screening were compared with the presence of pulmonary TB. Survival status at 12 months was recorded. Associations with CD4 count, BMI, haemoglobin, disease severity, and mortality were analysed. ResultsThe plasma levels of the biomarkers were significantly higher in TB-positive (n:21) compared to TB-negative (n:122) subjects. WHO symptoms, increased neopterin (>10 nmol/L) and CRP (>10 mg/L) showed similar sensitivity and different specificity, with increased CRP showing higher and increased neopterin lower specificity. The three markers were inversely correlated to haemoglobin and to CD4, and CRP levels inversely correlated to BMI. The markers were also significantly higher in individuals with subsequent mortality and in individuals with higher mycobacterial load in sputum according to Xpert results (IP-10 and CRP). ConclusionThis study showed significant associations of the biomarkers analysed with TB infection and mortality, that could have potential clinical relevance. Biomarker levels may be included in operational research on TB screening and diagnosis.

Serum IP-10 levels and increased DPPIV activity are linked to circulating CXCR3+ T cells in cholestatic HCV patients.

Background & aimsSerum interferon-gamma-inducible protein-10 (IP-10) is elevated in cholestatic liver diseases and predicts response to antiviral therapy in patients with chronic hepatitis C virus (HCV) infection. Dipeptidylpeptidase 4 (DPPIV) cleaves active IP-10 into an inactive form, which inhibits recruitment of CXCR3+ T cells to the liver. In this study the link between IP-10 levels, DPPIV activity in serum and CXCR3+ T cells is analysed in cholestatic and non-cholestatic liver patients.MethodsIn serum DPPIV activity (by enzymatic assay), IP-10 (by ELISA) and bile acids (BA) (by enzymatic assay) were analysed in 229 naive HCV genotype (GT) 1 patients and in 16 patients with cholestatic liver disease. In a prospective follow-up (FU) cohort of 27 HCV GT 1 patients peripheral CD3+CXCR3+, CD4+CXCR3+ and CD8+CXCR3+ cells were measured by FACS.ResultsIn 229 HCV patients serum IP-10 levels correlated positively to DPPIV serum activity. Higher IP-10 levels and DPPIV activity were detected in cholestatic and in cirrhotic HCV patients. Increased IP-10 serum levels were associated with therapeutic non-response to antiviral treatment with pegylated-interferon and ribavirin. In the HCV FU cohort elevated IP-10 serum levels and increased BA were associated with higher frequencies of peripheral CD3+CXCR3+, CD4+CXCR3+ and CD8+CXCR3+ T cells. Positive correlation between serum IP-10 levels and DPPIV activity was likewise validated in patients with cholestatic liver diseases.ConclusionsA strong correlation between elevated serum levels of IP-10 and DPPIV activity was seen in different cholestatic patient groups. Furthermore, in cholestatic HCV patients a functional link to increased numbers of peripheral CXCR3+ immune cells could be observed. The source of DPPIV release in cholestatic patients remains open.

The association between serum cytokine and chemokine levels and antiviral response by entecavir treatment in chronic hepatitis B patients.

Although nucleoside/nucleotide analogue therapy is thought to suppress chronic hepatitis B (CHB) via regulation of inflammatory cytokines/chemokines, the mechanism is still unclear. In this study, serum cytokine/chemokine levels were measured in CHB patients treated with entecavir, and the association with antiviral response was analysed. A total of 78 Japanese patients with CHB were enrolled, and serum cytokine/chemokine levels were measured at baseline and at 12, 24 and 48 weeks of entecavir treatment using the MULTIPLEX kit. Antiviral response to entecavir treatment was significantly associated with hepatitis B surface antigen (HBsAg) titre and serum interferon-gamma-inducible protein 10 (IP-10) level (12w; P=0.0002; OR=0.020 [95% CI 0.002, 0.156], P=0.003; OR=0.042 [95% CI 0.005, 0.336], respectively). HBe-positive patients whose serum macrophage-derived chemokine (MDC) level was lower (<582.83 pg/ml) and IP-10 level was higher (≥1,323.13 pg/ml) achieved hepatitis B e antigen (HBeAg) loss earlier than those who remained HBeAg-positive (P=0.044). HBsAg reduction by entecavir treatment was significantly associated with higher initial tumour necrosis factor-alpha (TNF-α) level (≥15.20 pg/ml) and higher alanine aminotransferase level (≥73 IU/l; P=0.009; OR=18.460 [95% CI 2.044, 166.709], P=0.022; OR=7.709 [95% CI 1.341, 44.327], respectively). Results of the present study indicate that changes in cytokine/chemokine levels following entecavir therapy are associated with response to antiviral therapy in CHB patients. Monitoring of serum cytokine/chemokine levels could be useful for predicting reduction of HBV DNA and HBsAg and HBe seroconversion.

Associations of Self-Reported Sleep Quality with Circulating Interferon Gamma-Inducible Protein 10, Interleukin 6, and High-Sensitivity C-Reactive Protein in Healthy Menopausal Women.

IntroductionSleep disturbance is very common in menopausal women and poor sleep quality has been linked to systemic inflammation. However, the impact of poor sleep quality on health outcomes of menopausal women remains unclear. This study evaluated the relationships between sleep quality and inflammation in menopausal women.Participants and designThis cross-sectional study enrolled 281 healthy women aged 45 to 60 years. The Pittsburgh Sleep Quality Index (PSQI) was used to measure quality of sleep. Multiplex assays were used to measure the levels of 9 cytokines in morning fasting plasma samples. Other variables measured in this study included clinical characteristics and high-sensitivity C-reactive protein (hs-CRP).SettingThe study was performed at a medical center.ResultsThe 281 participants comprised 79 (28%) perimenopausal women and 202 (72%) postmenopausal women. Global PSQI scores were positively correlated with plasma hs-CRP levels (P = 0.012) and were marginally associated with interferon gamma-inducible protein-10 (IP10), interleukin 6 (IL6), and macrophage inflammatory protein-1beta (MIP-1β) levels. After adjusting for age, body mass index, menopause duration, and follicle stimulating hormone, multiple linear regression analysis revealed that high PSQI scores and sleep efficiency < 65% were associated with elevated plasma levels of hs-CRP, IP10, and IL6. In addition, sleep duration < 5 hours was associated with high hs-CRP levels.ConclusionOur data show that poor sleep quality and low sleep efficiency are associated with elevated levels of circulating inflammatory factors IP10, IL6 and hs-CRP and that short sleep duration is associated with high levels of hs-CRP in menopausal women. These findings provide novel evidence that poor sleep quality is linked to low-grade systemic inflammation in menopausal women.

Pretreatment serum levels of interferon-gamma-inducible protein-10 are associated with virologic response to telaprevir-based therapy

AimTelaprevir (TVR) remarkably improves the efficacy of interferon treatment for chronic hepatitis C. Interleukin-28B (IL28B) genotype and interferon-gamma-inducible protein-10 (IP-10) level predict virologic response to peg-interferon (Peg-IFN)/ribavirin (RBV) therapy. We aimed to investigate the usefulness of pretreatment serum IP-10 levels and IL28B genotyping in predicting sustained virologic response (SVR) to TVR-based triple therapy. MethodsIn this multi-center study, patients infected with hepatitis C virus genotype 1 with high viral load (⩾5.0logIU/mL) were treated with TVR for 12weeks and Peg-IFN/RBV for 24weeks in Japan. IL28B genotype, serum IP-10 levels, other clinical parameters, and drug dosages were assessed before treatment. ResultsWe included 121 patients who were treated with TVR for at least 8weeks and Peg-IFN/RBV for 24weeks. The median IP-10 levels were significantly lower in rapid virologic response (RVR) or SVR in the IL28B non-TT genotype group, with no significant difference in the TT genotype group. RVR rates were significantly lower in the group with higher serum IP-10 levels (>450pg/mL). In the non-TT IL28B genotype group, RVR and SVR rates were significantly lower in the group with higher IP-10 levels. SVR rates in the group with lower IP-10 levels (<450pg/mL) increased to 82% for those showing RVR, but reduced to 27% in the group with higher IP-10 levels for those not showing RVR. ConclusionsDetermination of serum IP-10 levels before treatment could be useful for predicting favorable virologic response to TVR-based triple therapy, especially in patients with IL28B non-TT genotype.

Interferon-Gamma Inducible Protein-10(IP10) as a Predictor of Early Virologic Response in Chronic Hepatitis C Infected Egyptian Patients Stratified for the Interleukin-28B rs12979860 Genotyp

Background and study aim: Single nucleotide polymorphisms near Interleukin 28B are strongly associated with favourable treatment response of chronic hepatitis C such as, the homozygous CC at markers129798Session [CurrentTestPartID]. Interferon-Gamma Inducible Protein-10 (IP-10) can be produced by a variety of cells, including hepatocytes. Pre-treatment plasma levels of IP10 are elevated in patients chronically infected with hepatitis C virus of genotypes 1 or 4 who do not achieve early virological response(EVR)to treatment. The aim of this study was to evaluate the rule of adding IP-10 to IL28B rs129798Session [CurrentTestPartID] Genotype in predicting EVR . Patients and methods: The study enrolled 78 naive chronic HCV patients who have criteria that met the pegylated interferon plus ribavirin (pegIFN-RBV) treatment for chronic hepatitis C virus (HCV) .IP-10 assay and single nucleotide polymorphisms of the IL28B genotype were performed. Results: Patients with EVR was younger than those without EVR with statistically significant different. Patients with EVR had less elevated liver enzyme ,low viral load and fasting blood sugar than those without EVR with statistically significant difference. 100% patients with out early virological response had A2 activity while 31.7% only of patients with EVR had A2 activity with statistically significant difference .Patients with EVR showed lower level of IP10 and 81.7% of them had CC allele genotype with statistically significant difference when compared to patients without EVR. Patients with CC genotype were associated with lower level of IP10, ALT, AST and also low viral load. Patients with low level of IP10 had lower levels of liver enzyme. Cut off level of IP 10 was<Session [CurrentTestPartID]5 pico gram/ml; at this cut off value sensitivity was=100%, specificity was= 96.7% and area under the curve (AUC)= 0.99. Conclusion: IP10 level was lower among responder group. IL28 genotype CC was significantly higher in responders when compared with non responders. .Patients with CC genotype were associated with lower level of IP10 and liver enzymes. Patients with CC genotype were associated also with low viral load.

Alanine aminotransferase, HCV RNA levels and pro-inflammatory and pro-fibrogenic cytokines/chemokines during acute hepatitis C virus infection.

BackgroundThis study assessed the association of alanine-aminotransferase (ALT) and hepatitis C virus (HCV) RNA levels with pro-inflammatory and pro-fibrogenic cytokines and chemokines during acute HCV infection to provide further insight into the potential HCV immunopathogenesis.MethodsParticipants in the ATAHC study, a prospective study of recent HCV infection, with detectable HCV RNA at the time of HCV detection were included. Plasma levels of 27 cytokines and chemokines were measured and their correlation with ALT and HCV RNA levels were assessed. Log10 transformed cytokines and ALT values were used in the analysis.ResultsAmong 117 individuals, the plasma levels of interferon-gamma inducible protein-10 (IP-10) and macrophage inflammatory protein-1beta (MIP-1β) were positively correlated with ALT levels (IP-10: r = 0.42, P < 0.001; MIP-1β: r = 0.29, P = 0.001) and HCV RNA levels (IP-10: rs = 0.44, P < 0.001; MIP-1β: rs = 0.43, P < 0.001). Using linear regression, after adjusting for sex, age, infection duration, symptomatic infection, HIV co-infection, interferon-lambda rs12979860 genotype, HCV genotype, and assay run, higher ALT levels (β = 0.20; 95 % CI: 0.07, 0.32; P = 0.002) and HCV RNA levels >400,000 IU/mL (vs. <8,500 IU/mL; β = 0.16; 95 % CI: 0.03, 0.28; P = 0.014) were independently associated with higher IP-10 levels. HCV RNA levels >400,000 IU/mL (vs. <8,500 IU/mL; β = 0.16; 95 % CI: 0.01, 0.31; P = 0.036) were associated with higher MIP-1β levels.ConclusionsDuring acute HCV infection, high ALT and HCV RNA levels were associated with increased IP-10 levels, while high HCV RNA levels were also associated with increased MIP-1β levels. These data suggest that IP-10 and MIP-1β may have a role in HCV immuno-pathogenesis starting early in acute HCV infection.Electronic supplementary materialThe online version of this article (doi:10.1186/s12985-016-0482-x) contains supplementary material, which is available to authorized users.

Interferon gamma inducible protein-10 in the diagnosis of paediatric tuberculosis infection in a low TB incidence country.

To assess interferon-gamma inducible protein 10 (IP-10) as a diagnostic marker for tuberculous infection in children, particularly in relation to its differential diagnostic performance in young children. A case-control study was conducted among 161 children and adolescents (mean age 6.3 years ± standard deviation 1.7; males n = 79, 49%). Fifty-four (33.5%) had active TB, 53 (33%) had latent tuberculous infection (LTBI), and 54 (33.5%) were non-LTBI controls. Unstimulated IP-10 levels did not differ between groups (P > 0.05 for all comparisons). TB-specific antigen stimulated IP-10 levels were more profoundly increased in infected groups than in controls (P < 0.001 for all comparisons). None of the IP-10-based diagnostic indexes demonstrated the ability to discriminate active disease from LTBI. A value of IP-10 ⩾ 1222 pg/ml had 83.3% sensitivity, 79.6% specificity, 80.4% positive predictive value and 82.7% negative predictive value for the diagnosis of LTBI. IP-10 based indexes demonstrated a trend towards better performance in the population group aged <5 years. The IP-10 assay could be useful in improving the diagnosis of LTBI in patients aged <5 years in combination with existing interferon-gamma release assays.

Elevated Urinary T Helper 1 Chemokine Levels in Newly Diagnosed Hypertensive Obese Children

Objective:Increasing evidence suggests that T helper (Th) cells play a significant role in the pathogenesis of hypertension. The aim of this study was to evaluate the effect of obesity and anti-hypertensive treatment on urinary Th1 chemokines.Methods:The study groups consisted of three types of patients: hypertensive obese, healthy, and non-hypertensive obese. Pre-treatment and post-treatment samples of the hypertensive obese group and one sample from the other two groups were evaluated for urinary chemokine: regulated on activation, normal T cell expressed and secreted (RANTES), interferon-gamma-inducible protein 10 (IP10), and monokine induced by interferon-gamma (MIG). In the hypertensive obese group, urine microalbumin: creatinine ratio was examined before and after treatment. We recommended lifestyle changes to all patients. Captopril was started in those who could not be controlled with lifestyle changes and those who had stage 2 hypertension.Results:Twenty-four hypertensive obese (mean age 13.1), 27 healthy (mean age 11.2) and 22 non-hypertensive obese (mean age 11.5) children were investigated. The pre-treatment urine albumin: creatinine ratio was positively correlated with pre-treatment MIG levels (r=0.41, p<0.05). RANTES was significantly higher in the pre-treatment hypertensive and non-hypertensive obese group than in the controls. The urinary IP10 and MIG levels were higher in the pre-treatment hypertensive obese group than in the non-hypertensive obese. Comparison of the pre- and post-treatment values indicated significant decreases in RANTES, IP10, and MIG levels in the hypertensive obese group (p<0.05).Conclusion:Th1 cells could be activated in obese hypertensive children before the onset of clinical indicators of target organ damage. Urinary RANTES seemed to be affected by both hypertension and obesity, and urinary IP10 and MIG seemed to be affected predominantly by hypertension.

Serum levels of leptin and IP-10 in preeclampsia compared to controls.

Preeclampsia (PE) is a pregnancy-specific syndrome with a complex, yet elusive, etiology. The production of a variety of factors probably implicated in diverse pathways may trigger endothelial dysfunction leading to PE pathogenesis. The aim of the present study was to investigate and compare the concentrations of leptin and interferon-gamma-inducible protein-10 (IP-10), factors characterized by inflammatory, immunomodulatory and angiogenic activities, and to evaluate their possible interaction in women with normotensive pregnancy and PE. The study was carried out on a total of 58 pregnant women, 29 women with PE and 29 controls. Serum leptin and IP-10 levels were determined by enzyme-linked immunosorbent assay. Serum leptin levels were significantly increased in women with PE compared to controls and this difference was stronger in women with severe PE (p < 0.001). Although IP-10 serum concentrations were elevated in our preeclamptic women, this difference was not statistically significant. No correlation was found between leptin and IP-10. The results of the present study support a significant role of leptin in PE; however, this association was independent from serum IP-10 levels, suggesting that there is no crucial interplay between these two proteins in PE.

Association ofInterferon-Gamma Inducible Protein 10Polymorphism with Treatment Response to Pegylated Interferon in HBeAg-Positive Chronic Hepatitis B

Interferon-gamma inducible protein 10 (IP-10) plays an important role in the clinical outcome of chronic hepatitis B (CHB). This study aimed to investigate the association between single nucleotide polymorphisms (SNPs) G-201A of the IP-10 gene and treatment response to pegylated interferon (PEG-IFN) in patients with hepatitis B e antigen (HBeAg)-positive CHB. We retrospectively analysed data of patients with HBeAg-positive CHB treated with PEG-IFN for 48 weeks. Virological response (VR) was defined as HBeAg clearance and HBV DNA <2,000 IU/ml at 24 weeks post-treatment. The SNPs G-201A, IFNL3 (rs12979860) and HLA-DPA1 (rs3077) were assessed. Among 107 patients, VR was achieved in 45 (42.1%) patients. Hepatitis B surface antigen (HBsAg) clearance and decline (<100 IU/ml) were observed in 10 (9.3%) and 22 (20.6%) patients, respectively. The distribution of GG, GA and AA genotypes of G-201A was 76.6%, 19.6% and 3.7%, respectively. Patients with GG genotype, compared to those with non-GG genotype, achieved higher VR rate (48.8% versus 19.2%; P=0.011), decreased HBsAg (25.6% versus 4.0%; P=0.019), and demonstrated a trend in HBsAg clearance (11.0% versus 4%; P=0.294). Patients with GG genotype had more rapid HBsAg decline and higher baseline serum IP-10 levels than those with non-GG genotype (432.2 ±339.0 versus 257.3 ±145.7 pg/ml; P=0.028). SNPs rs12979860 and rs3077 were not associated with VR. Logistic regression analysis suggested that SNP G-201A was an independent predictor of VR (odds ratio 3.81, 95% CI 1.31, 11.12; P=0.014). Data from this study demonstrated for the first time that IP-10 polymorphism is independently associated with treatment response to PEG-IFN in patients with HBeAg-positive CHB.

Serum Fractalkine and Interferon-Gamma Inducible Protein-10 Concentrations Are Early Detection Markers for Acute Renal Allograft Rejection

ObjectiveThe aims of this study were to determine if characterization of serum concentrations of interferon-gamma inducible protein-10 (IP-10), fractalkine, and their receptors (CXCR3 and CX3CR1) were predictive of acute allograft rejection in kidney transplant recipients. MethodsKidney transplant recipients (n = 52) were enrolled in this study and divide into either the acute rejection (AR, n = 15) or non–acute rejection (NAR, n = 35) groups. Serum samples from recipients were collected 1 day prior to transplantation and on days 1, 3, 5, 7, and 9 post-transplantation. The accuracy of chemokine concentrations for predicting acute rejection episodes was evaluated using receiver operator characteristic (ROC) curves. ResultsAR was diagnosed in 15 patients based on histologic changes to renal biopsies. AR patients had significantly higher serum fractalkine, CXCR1, IP-10, and CXCR3 levels compared to levels observed in the NAR group and healthy controls. Fractalkine and IP-10 had the largest area under the ROC curve at 0.86 (95% confidence interval: 0.77–0.96). Following steroid therapy, chemokine levels decreased, which may serve to predict the therapeutic response to steroid therapy. ConclusionMeasuring serum levels of fractalkine, IP-10, and their receptors (especially the fractalkine/IP-10 combination) may serve as a noninvasive approach for the early diagnosis of renal allograft rejection.

Development of a highly sensitive liquid chromatography/tandem mass spectrometry method to quantify total and free levels of a target protein, interferon-gamma-inducible protein-10, at picomolar levels in human serum.

Liquid chromatography/tandem mass spectrometry (LC/MS/MS) assays are increasingly being used for absolute quantitation of proteins due to high specificity and low cost. However, the major challenge for the LC/MS method is insufficient sensitivity. This paper details the strategies developed to maximize the sensitivity from aspects of chromatography, mass spectrometry, and sample preparation to achieve a highly sensitive LC/MS method. The method is based on the LC/MS/MS measurement of a surrogate peptide generated from trypsin digestion of interferon-gamma-inducible protein-10 (IP-10). The sample preparation strategy involved selectively extracting IP-10 and removing high-abundance serum proteins through acidified protein precipitation (PPT). It was revealed in this work that these high-abundance serum proteins, if not separated from the protein of interest, could cause significant ionization saturation and high background noise in selected reaction monitoring (SRM), leading to a 100-fold higher lower limit of quantification (LLOQ). Our method demonstrated that the acidified PPT could be optimized to selectively extract the protein of interest with full recovery of 97% to 103%, while the high-abundance serum proteins could be effectively removed with minimal matrix effect of 90% to 93%. For the first time, a highly sensitive LC/MS method with a LLOQ of 31.62 pM for the quantitation of IP-10 has been achieved, which is a 100-fold improvement over the generic method. The described method offers excellent sensitivity with advantages of being antibody reagent independent and leads to significant cost and time savings. It has been successfully employed to determine both total and free IP-10 levels in human serum samples. This method development strategy may also be applied to other small proteins.

Su1003 High Pre-Treatment Serum Interferon-Gamma-Inducible Protein 10 (IP-10) Levels Is Associated With HBsAg Seroconversion After Peginterferon Therapy in HBeAg-Positive and HBeAg-Negative Chronic Hepatitis B

Introduction: Serum levels of interferon-gamma inducible protein 10 (IP-10) are a marker for immune activity and may be associated with an increased probability of HBeAg loss after pegylated interferon (PegIFN) therapy. The aim of this study is to compare the chronological change of serum IP-10 levels in chronic hepatitis B patients with and without HBsAg seroconversion after PegIFN therapy. Materials and Methods: Sixty chronic hepatitis B patients (HBeAg positive (n=30) and HBeAg negative (n=30)) were recruited and treated with PegIFN for 48 weeks. Demographic and clinical data of the patients were recorded. Serum IP-10 was measured at baseline, on-treatment weeks 12, 24 and 48 and week 72 post-treatment. Results: Forty-four males and 16 female chronic hepatitis B patients were included. The median age of patients was 34 years, and 7 of 60 patients achieved HBsAg seroconversion at week 96 post-treatment. Baseline characteristics including age, sex, ALT, HBV DNA level and HBsAg level were not statistical different between the two groups (Table 1). The mean level of serum IP-10 at baseline was 751±588 and 250±293 pg/ml in patients with and without HBsAg seroconversion, respectively. At baseline and week 48, IP-10 level were significantly higher in patients with HBsAg seroconversion than patient without HBsAg seroconversion (Table 2). A cut off value of baseline serum IP-10 level of less than 200 pg/ ml predicted no one achieving HBsAg seroconversion at week 96 post-treatment (100%, 32/32). Whereas, up to 25% (7/28) of CHB patients with a baseline serum IP-10 level more than 200 pg/ml achieved HBsAg seroconversion at week 96 after PegIFN therapy. The odds ratio for HBsAg seroconversion at week 96 after PegIFN therapy was 1.37 for every 100 pg/ml positive increment of baseline IP-10 (OR 1.37; 95% CI, 1.07-1.59; P<0.01). The dynamic change of the post-PegIFN treatment serum IP-10 level was significantly different among two groups of patients. In the HBsAg seroconversion group, the serum IP-10 level which was not statistically significant changed during treatment, however, the IP-10 was significantly decreased in week 72 post-treatment. On the other hand, the IP-10 level tended to significantly increase during the treatment period in chronic HBV patients without seroconversion after Peg-IFN treatment (Figure 1). Conclusions: Pre-treatment serum IP-10 was higher in patients who achieved HBsAg seroconversion at week 96 after peginterferon therapy. This pilot study showed that a cut off value of 200 pg/ml serum IP-10 level predicted the outcome of treatment with Peg-IFN in chronic hepatitis B patients. Table 1. Comparison of demographic, baseline ALT, HBV DNA and HBsAg level between patients with and without HBsAg seroconversion at week 96.