Mechanism of action of DSP-7888 (adegramotide/nelatimotide) Emulsion, a peptide-based therapeutic cancer vaccine with the potential to turn up the heat on non-immunoreactive tumors

Background Wilms’ tumor 1 (WT1) is highly expressed in various solid tumors and hematologic malignancies. DSP-7888 (adegramotide/nelatimotide) Emulsion is an investigational therapeutic cancer vaccine comprising three synthetic epitopes derived from WT1. We evaluated the mechanism of action of DSP-7888 Emulsion, which is hypothesized to induce WT1-specific cytotoxic T lymphocytes (CTLs) and helper T lymphocytes (HTLs). Methods The ability of nelatimotide and adegramotide to induce WT1-specific CD8+ T cells and CD4+ T cells was assessed in human peripheral blood mononuclear cells (PBMCs). The ability of DSP-7888 Emulsion to induce WT1-specific CTLs in vivo was assessed using human leukocyte antigen-I (HLA-I) transgenic mice. To assess how adegramotide, the helper peptide in DSP-7888 Emulsion, enhances WT1-specific CTLs, HLA-I transgenic mice were administered DSP-7888 or nelatimotide-only Emulsion. Interferon-gamma secretion under antigen stimulation by splenocytes co-cultured with or without tumor cells was then quantified. The effects of combination treatment with DSP-7888 Emulsion and an anti–programmed cell death protein 1 (PD-1) antibody on tumor volume and the frequency of tumor-infiltrating WT1-specific T cells were assessed in HLA-I transgenic mice implanted with WT1 antigen-positive tumors. Result The peptides in DSP-7888 Emulsion were shown to induce WT1-specific CTLs and HTLs in both human PBMCs and HLA-I transgenic mice. Unlike splenocytes from nelatimotide-only Emulsion-treated mice, splenocytes from DSP-7888 Emulsion-treated mice exhibited high levels of interferon-gamma secretion, including when co-cultured with tumor cells (AU)

The “time-response” effect of Wenyang Pingchuan Formula on miR-19a in asthmatic mice experiment

Background: Wenyang Pingchuan Formula (WPCF) is an empirical formula for the treatment of acute childhood asthma. However, the “time-effect” relationship of this prescription is not clear. This paper explores the relationship between Janus activated kinase signal transducer and activator of transcriptions (JAK/STAT), nuclear factor-κB (NF-κB), and microRNA (miR-19a), and also preliminarily determines the best time-effect relationship of WPCF in reducing the airway inflammation in asthmatic mice.Method: 80 BALB/c mice were randomly divided into four groups: control (CON) group, model (MDL) group, dexamethasone (DEX) group, and WPCF group. MDL group was established through intraperitoneal injection of 10% ovalbumin (OVA) and Al(OH)3 solution and the inhalation of aerosolized 5% OVA solution. Enzyme-linked immunosorbent assay (ELISA), real-time PCR and Western blot were conducted to determine the levels of interleukin (IL)-4, IL-13, interferon-γ (IFN-γ) in bronchoalveolar lavage fluid (BALF), contents of miR-19a mRNA and STAT6, phosphorylated signal transducers and activators of transcription 6 (p-STAT6), p65, phosphorylated p65 (p-p65), suppressors of cytokine signaling 1 (SOCS1), and tumor necrosis factor α-induced protein-3 (Tnfaip3) proteins after 7 and 28 days of intervention respectively.Results: Significant down-regulation of IL-4 and IL-13 expression (P<0.05) and up-regulation of IFN-γ expression (P<0.05) in BALF have been observed for WPCF group compared with the MDL group. The significant down-regulation of miR-19a mRNA and STAT6, p-STAT6, p65, p-p65 proteins (P<0.05) and up-regulation of SOCS1 and Tnfaip3 proteins (P<0.05) in BALF was also observed for WPCF group compared to the MDL group. During the experiment, the weight of the mice in DEX group significantly decreased (P<0.05) compared with the other groups.Conclusions: WPCF could restore Th1/Th2 balance (AU)

IFN-γ affects pancreatic cancer properties by MACC1-AS1/MACC1 axis via AKT/mTOR signaling pathway

BackgroundMetastasis-related in colon cancer 1 (MACC1) is highly expressed in a variety of solid tumours, but its role in pancreatic cancer (PC) remains unknown. Interferon gamma (IFN-γ) affecting MACC1 expression was explored as the potential mechanism following its intervention.MethodsExpressions of MACC1 treated with IFN-γ gradient were confirmed by quantitative real-time PCR (qRT-PCR) and western blot (WB). Proliferation, migration, and invasion abilities of PC cells treated with IFN-γ were analysed by CCK8, EDU, colony formation, Transwell (with or without matrix gel) and wound-healing assays. Expression of antisense long non-coding RNA of MACC1, MACC1-AS1, and proteins of AKT/mTOR pathway, (pho-)AKT, and (pho-)mTOR was also assessed by qRT-PCR and WB. SiRNA kit and lentiviral fluid were conducted for transient expression of MACC1 and stable expression of MACC1-AS1, respectively. Rescue assays of cells overexpressing MACC1-AS1 and of cells silencing MACC1 were performed and cellular properties and proteins were assessed by the above-mentioned assays as well.ResultsIFN-γ inhibited MACC1 expression in a time- and dose-dependent manner; 100 ng/mL IFN-γ generally caused downregulation of most significant (p ≤ 0.05). In vitro experiments revealed that IFN-γ decreased cellular proliferation, migration, and invasion abilities and downregulated the expression of pho-AKT and pho-mTOR (p ≤ 0.05). Conversely, overexpression of MACC1-AS1 upregulated pho-AKT and pho-mTOR proteins, and reversed cellular properties (p ≤ 0.05). Rescue assays alleviated the above changes of pho-AKT/ mTOR and cellular properties.ConclusionIFN-γ affected PC properties by MACC1-AS1/MACC1 axis via AKT/mTOR signaling pathway, which provides novel insight for candidate targets for treating PC.

Gen 15 estimulado por interferón, nexo común del daño vascular en la hipertensión, la obesidad y los aneurismas aórticos / Interferon-stimulated gene 15, a common link of vascular damage in hypertension, obesity and aortic aneurysms

La hipertensión y la obesidad son importantes problemas de salud en todo el mundo con notables consecuencias sobre la morbilidad y la mortalidad. De hecho, tanto la hipertensión como la obesidad son importantes factores de riesgo para el desarrollo de enfermedades cardiovasculares. La disfunción endotelial, el remodelado vascular y las alteraciones en la mecánica vascular son aspectos comunes del daño vascular en la hipertensión, la obesidad y los aneurismas. Durante las últimas décadas, se ha demostrado la importancia de la inflamación de bajo grado en el daño vascular asociado a las enfermedades cardiovasculares. Dicha inflamación se caracteriza por la acumulación de células inflamatorias en la vasculatura, así como por el aumento de citoquinas proinflamatorias locales y circulantes. Por tanto, la identificación de nuevos mediadores inflamatorios implicados en dicho daño se ha convertido en un área de investigación muy importante. El interferón-γ (IFNγ) o el factor de necrosis tumoral-α (TNFα) son importantes citoquinas implicadas en el daño vascular asociado a la hipertensión. Además, se acepta que el TNFα es un mediador clave implicado en la resistencia a la insulina y el daño vascular observado en la obesidad. Ambas citoquinas inducen la expresión del gen 15 estimulado por interferón (ISG15), una proteína similar a la ubiquitina que induce una modificación postraducional reversible (ISGilación) y que también puede secretarse como forma libre. Las funciones de ISG15 están principalmente relacionadas con la respuesta inmune frente a infecciones, sin embargo, podría ser también una interesante nueva diana del daño cardiovascular.(AU)

Hypertension and obesity are major health problems worldwide with significant consequences on morbidity and mortality. In fact, both hypertension and obesity are important risk factors for the development of cardiovascular diseases. Endothelial dysfunction, vascular remodeling, and alterations in vascular mechanics are common aspects of vascular damage in hypertension, obesity, and aneurysms. During the last decades, the importance of low-grade inflammation in vascular damage associated with cardiovascular diseases has been demonstrated. This inflammation is characterized by the accumulation of inflammatory cells in the vasculature, as well as by the increase of local and circulating pro-inflammatory cytokines. Therefore, the identification of new inflammatory mediators involved in this damage has become a very important area of research. Interferón-γ (IFNγ) or tumor necrosis tumoral-α (TNFα) are important cytokines involved in the vascular damage associated with hypertension. Furthermore, it is accepted that TNFα is a key mediator involved in insulin resistance and vascular damage observed in obesity. Both cytokines induce the expression of interferon-stimulated gene 15 (ISG15), a protein similar to ubiquitin that induces a reversible post-translational modification (ISGylation) and that can also be secreted as a free form. The functions of ISG15 are mainly related to the immune response against infections, however, it could also be an interesting new target for cardiovascular damage.(AU)

Mechanisms of primary and acquired resistance to PD-1/PD-L1 blockade and the emerging role of gut microbiome

As a very promising immunotherapy, PD-1/PD-L1 blockade has revolutionized the treatment of a variety of tumor types, resulting in significant clinical efficacy and lasting responses. However, these therapies do not work for a large proportion of patients initially, which is called primary resistance. And more frustrating is that most patients eventually develop acquired resistance after an initial response to PD-1/PD-L1 blockade. The mechanisms that lead to primary and acquired resistance to PD-1/PD-L1 inhibition have remained largely unclear. Recently, the gut microbiome has emerged as a potential regulator for PD-1/PD-L1 blockade. This review elaborates on the current understanding of the mechanisms in terms of PD-1 related signaling pathways and necessary factors. Moreover, this review discusses new strategies to increase the efficacy of immunotherapy from the perspectives of immune markers and gut microbiome (AU)

Echinococcus multilocularis induces surface high expression of inhibitory killer immunoglobulin-like receptor on natural killer cells

Alveolar echinococcosis (AE) is a malignant and fatal parasitic disease caused by the larvae of Echinococcus multilocularis (E. multilocularis), which inhibits the activity and proliferation of natural killer (NK) cells. In this study, the functional alteration of hepatic NK cells and their related molecules were studied. The AE-infected patient’s tissue was fixed with formalin, embedded in paraffin, and stained with Masson’s trichrome or hematoxylin and eosin (H&E). Single cells from AE-infected patient or E. multilocularis-infected mice were blocked with Fc-receptor (FcR), and stained with monoclonal antibodies, including CD16, CD56, CD3, KIR2DL1, granzyme B, perforin, Interferon gamma (IFN-γ), and tumor necrosis factor-α (TNFα) or isotype control, to measure molecules and cytokines of NK cells and analyzed by flow cytometry. The Sirius red staining was used to quantitate hepatic fibrosis by calculating quantitative collagen deposition. AE can adjust both the number of hepatic CD56+ NK cells and its KIR2DL1 expression processes. Moreover, the overexpression of KIR2DL1 in NK cells could downregulate the functioning of immune cells in the liver area close to parasitic lesions. The number and dysfunction of NK cells in E. multilocularis infection could be related to the molecule dynamics of cell surface inhibitory receptor Ly49A, leading to hepatic damage and progression of fibrosis. This study illustrated significant increase in hepatic fibrogenesis and apparent upregulation of hepatic CD56+ NK cell population and its KIR2DL1 expression in AE-infected patients. This opposite variation might be related to the impaired NK cells functioning, such as granzyme B, IFN-γ, and TNF-α secretion. In addition, the cell surface inhibitory receptor Ly49A was related to the intracellular cytokine secretion functions of NK cells (AU)

Fungal co-infection in COVID-19 patients: should we be concerned? / Coinfección fúngica en pacientes con COVID-19: ¿debemos preocuparnos?

Critically ill COVID-19 patients have higher pro-inflammatory (IL-1, IL-2, IL-6, tumor necrosis alpha) and anti-inflammatory (IL-4, IL-10) cytokine levels, less CD4 interferon-gamma expression, and fewer CD4 and CD8 cells. This severe clinical situation increases the risk of serious fungal infections, such as invasive pulmonary aspergillosis, invasive candidiasis or Pneumocystis jirovecii pneumonia. However, few studies have investigated fungal coinfections in this population. We describe an update on published reports on fungal coinfections and our personal experience in three Spanish hospitals. We can conclude that despite the serious disease caused by SARS-CoV-2 in many patients, the scarcity of invasive mycoses is probably due to the few bronchoscopies and necropsies performed in these patients because of the high risk in aerosol generation. However, the presence of fungal markers in clinically relevant specimens, with the exception of bronchopulmonary colonization by Candida, should make it advisable to early implement antifungal therapy

Los pacientes gravemente enfermos con COVID-19 presentan concentraciones más elevadas de citoquinas pro-inflamatorias (IL-1, IL-2, IL-6 y factor de necrosis tumoral alfa) y anti-inflamatorias (IL-4 e IL-10), menor expresión de interferón-gama y un número más bajo de células CD4 y CD8. Esta grave situación clínica aumenta el riesgo de padecer coinfecciones fúngicas, como la aspergilosis pulmonar invasora, la candidiasis invasora o la neumonía por Pneumocystis jirovecii. Sin embargo, pocos estudios han investigado las coinfecciones fúngicas en esta población. En esta revisión describimos una actualización de las publicaciones sobre coinfecciones fúngicas en esta población de pacientes y nuestra experiencia personal en tres hospitales españoles. Podemos concluir que a pesar de la grave enfermedad causada por el SARS-CoV-2 en muchos pacientes, la baja frecuencia de micosis invasoras se debe probablemente a las pocas broncoscopias y necropsias realizadas en estos pacientes debido al alto riesgo de producción de aerosoles. Sin embargo, la presencia de marcadores fúngicos en muestras clínicas relevantes, con la excepción de la colonización broncopulmonar por Candida, debería aconsejar la instauración precoz de una terapia antifúngica

Atopy can be an interfering factor in genetic association studies of beta-lactam allergy

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IFN-γ stimulation of dental follicle mesenchymal stem cells modulates immune response of CD4+ T lymphocytes in Der p1+ asthmatic patients in vitro

Background: House dust mite (Dermataphagoides pteronyssinus) is a widespread risk factor in the development of asthma. CD4+ T lymphocytes have an important role in the pathogenesis of allergic asthma by polarizing to Th2 cells. Objective: We aimed to evaluate the immunoregulatory effects of dental follicle mesenchymal stem cells with and without IFN-γ stimulation on peripheral blood mononuclear cells of house dust mite sensitive asthmatic patients, and compared those with Dexamethasone as a systemic steroid. Material and methods: PBMC of asthmatic patients and healthy individuals separately cultured with or without DF-MSCs in the presence and absence of IFN-γ or Der p1 or Dexamethasone for 72h. CD4+ T proliferation, cell viability, CD4+CD25+FoxP3+ Treg cell frequency and cytokine profiles of PBMC were evaluated via flow cytometry. Results: DF-MSCs suppressed proliferation of CD4+ T lymphocytes (pCDmix < 0.01, pDerp1 < 0.01, pIFN < 0.005) by increasing the number of FoxP3 expressing CD4 + CD25 + T regulatory cells (pCDmix < 0.005, pDerp1 < 0.01, pIFN < 0.001) and suppressed lymphocyte apoptosis (pCDmix < 0.05, pDerp1< 0.05, pIFN < 0.05), while Dexamethasone increased the apoptosis and decreased Treg cell frequency in asthmatic patients. IFN-γ stimulation increased the suppressive effect of DF-MSCs and also enhanced the frequency of FoxP3 expressing CD4+CD25 + T regulatory cells. The cytokine levels were regulated by DF-MSCs by reducing IL-4 cytokine levels (pCDmix < 0.01, pDerp1 < 0.05, pIFN < 0.05) and upregulating IFN-γ levels (pCDmix < 0.01, pDerp1< 0.05, pIFN < 0.005) in asthmatic patients. Conclusion: IFN-γ stimulated DF-MSCs were found to have a high modulatory effect on CD4 + T cell responses, while Dexamethasone had an apoptotic effect on CD4+ T cells in asthmatic patients. DF-MSCs may be a new cell-based therapy option for allergic diseases including asthma

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Evaluation of interleukin-12 receptor Beta1 and interferon gamma receptor 1 deficiency in patients with disseminated BCG infection

Introduction: Disseminated BCG infections among other complications of Bacillus Calmette-Guérin (BCG) vaccine are rare and have occurred in children with immunodeficiency disorders such as mendelian susceptibility to mycobacterial disease (MSMD) which could be due to defects in some elements of IL-12/IFN-γ axis. MSMD-causing mutations have been identified in 10 genes during the last two decades. Among them, mutations in the IL12Rβ1 and IFN gamma R1 genes constitute about 80% of recorded cases of MSMD syndrome. The aim of this study was to investigate IL-12RBeta1 and IFN- gammaR1 deficiencies in patients with disseminated BCG infection. Methods: This study was performed on 31 children with disseminated BCG infections who referred to children's medical center. Whole blood cell culture was performed in presence of BCG, IL-12 and IFN- gamma stimulators. The supernatants were assayed for IFN-gamma and IL-12p70 by ELISA method. In order to evaluate IL12Rbeta1 and IFN- gammaR1 receptors expression, flow cytometry staining was performed on the patients’ T-cells stimulated with PHA. Results: Flow cytometry staining of 31 Iranian patients with disseminated BCG infections with the average age of 43 months showed lack of the expression of IL-12RBeta1 and IFN- gamma R1 genes in PHA-T-cells of the nine and one patients, respectively in whom the incomplete production of IFN- gamma and IL-12 was reported by ELISA. Among these 10 patients, eight cases had related parents (80%). Conclusion: It is recommended that to avoid BCG complications, screening be performed for MSMD before BCG inoculation in individuals with positive family history of primary immunodeficiency diseases and inhabitants of areas with high frequency of consanguinity

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Single nucleotide polymorphisms of IL-2, but not IL-12 and IFN-y, are associated with increased susceptibility to chronic spontaneous urticaria

Background: A clear picture of interaction of Th1/Th2 cytokines in pathogenesis of chronic spontaneous urticaria (CSU), remains elusive. Impaired IFN-γ production and decreased levels of IL-2 have been reported. The aim of this study was to evaluate the association of Th1 cytokines; IL-2, IL-12 and IFN-γ polymorphisms with CSU. Methods: 90 patients with CSU and 140 age-sex matched subjects were included in this study. DNA samples were evaluated through PCR-SSP assay in order to detect single nucleotide polymorphisms of IL-12 (A/C -1188) or (rs3212227), IFN-γ (A/T UTR5644) or (rs2069717) and IL-2 (G/T -330 and G/T +166) or (rs2069762 and rs2069763). Results: G allele at -330 at promoter region of IL-2 gene was overrepresented in CSU. Heterozygotes (GT) at this locus and heterozygotes at +166 of IL-2 gene (GT) were more prevalent in CSU group. Additionally, the haplotype GT for loci -330 and +166 of IL-2 gene was powerfully associated with CSU (OR (95%CI) = 57.29 (8.43-112.7)). Conclusions: SNP at position -330 and +166 of IL-2 gene are differently expressed in CSU. The haplotype GT of IL-2 at -330 and +166 might confer vulnerability to a number of immunological disorders in Iranian region (AU)

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Tuberculosis diseminada en anciano de edad avanzada / Disseminated tuberculosis in elderly patients

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Sumario ejecutivo de la guía de práctica clínica sobre el uso de las pruebas de liberación de interferón-gamma para el diagnóstico de infección tuberculosa / Executive Summary of the Guidelines for the Use of interferon-gamma Release Assays in the Diagnosis of Tuberculosis Infection

El uso de las pruebas de liberación de interferón-gamma para el diagnóstico de la infección tuberculosa está muy generalizado en España. Sin embargo, no se ha alcanzado un consenso respecto a su aplicación en situaciones clínicas específicas. Con el fin de redactar una guía de empleo en la práctica clínica, un grupo de expertos que incluyó a especialistas en enfermedades infecciosas, enfermedades respiratorias, microbiología, pediatría y medicina preventiva, junto con un experto en metodología, efectuaron una búsqueda sistemática en la literatura, sintetizaron los resultados, calificaron la calidad de las evidencias y formularon recomendaciones de acuerdo con la metodología Grading of Recommendations of Assessment Development and Evaluations. Este documento es una guía basada en la evidencia para el empleo de las pruebas de liberación de interferón-gamma en el diagnóstico de la infección tuberculosa en pacientes en riesgo de padecer tuberculosis o en los que se sospeche enfermedad activa. La guía será aplicable tanto en atención primaria y especializada como en salud pública

Interferon-gamma release assays are widely used for the diagnosis of tuberculosis infection in Spain. However, there is no consensus on their application in specific clinical scenarios. To develop a guide-line for their use, a panel of experts comprising specialists in infectious diseases, respiratory diseases, microbiology, pediatrics and preventive medicine, together with a methodologist, conducted a systematic literature search, summarized the findings, rated the quality of the evidence, and formulated recommendations following the Grading of Recommendations of Assessment Development and Evaluations methodology. This document provides evidence-based guidance on the use of interferon-gamma release assays for the diagnosis of tuberculosis infection in patients at risk of tuberculosis or suspected of having active disease. The guidelines will be applicable to specialist and primary care, and public health

Association of Interleukin-2, but not Interferon-Gamma, single nucleotide polymorphisms with juvenile idiopathic arthritis

BACKGROUND: Cytokines, including interleukin-2 (IL-2) and interferon-gamma (IFN-γ), seem to play a role in the pathogenesis of juvenile idiopathic arthritis (JIA). The aim of this study was to investigate the associations of IL-2 and IFN-γ single nucleotide polymorphisms (SNPs) with susceptibility to JIA in an Iranian population. METHODS: enomic DNA of 54 Iranian patients with JIA and 139 healthy unrelated controls were typed for IL-2 (G/T at −330 and +166) as well as IFN-γ gene (A/T at +874), using polymerase chain reaction with sequence-specific primers method, and compared between patients and controls. RESULTS: A significantly higher frequency of the IL-2 −330 GG genotype (p < 0.01) was found in the JIA patients compared to the controls. However, the GT genotype at the same position was notably lower than in controls (p < 0.01). Moreover, IL-2 (−330, +166) GT haplotype was more frequent in patients with JIA in comparison with controls. No significant differences was observed between the two groups of case and control for IL-2 (G/T at +166) and IFN-γ (A/T at +874) SNPs. CONCLUSION: The results of the current study suggest that certain SNPs of IL-2 gene have association with individuals' susceptibility to JIA. However, further investigations are required to confirm the results of this study

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Improving T-cell assays for diagnosis of latent TB infection: Confirmation of the potential role of testing Interleukin-2 release in Iranian patients

BACKGROUND: Since gamma interferon release assays (IGRAs) cannot differentiate between active tuberculosis and latent tuberculosis infection (LTBI), development of rapid and specific diagnosis tools are essential for discriminating between active tuberculosis (TB) from LTBI. Both IGRAs are based on Mycobacterium tuberculosis-specific antigens, namely, early secretory antigenic target 6 (ESAT-6) and 10kDa culture filtrate (CFP-10). The aim of this study was to evaluate the potential value of IL-2 secretion by whole blood cells after stimulation with rESAT-6 and rCFP-10 for discriminating between active and latent tuberculosis. METHODS: Interleukin-2 and IFN-γ were measured after blood stimulation of 90 cases (30 with active TB, 30 with LTBI and 30 healthy controls) with recombinant ESAT-6 and CFP-10. Receiver operating characteristic (ROC) curve analysis was conducted to determine the best IL-2 and IFN-γ result thresholds in discriminating between cases with active or latent TB, and the corresponding sensitivity and specificity were recorded. RESULTS: The IFN-γ release assay demonstrated a good sensitivity and specificity (sensitivity 83-84% and specificity 92%) for diagnosis of tuberculosis. The discrimination performance of IL-2 assay (assessed by the area under ROC curve) between LTBI and patients with active TB were 0.75 and 0.8 following stimulation with rESAT-6 and rCFP-10, respectively. Maximum discrimination was reached at a cut-off of 11.6pg/mL for IL-2 after stimulation with recombinant rESAT-6 with 72% sensitivity and 79% specificity and 10.7pg/mL for IL-2 following stimulation with rCFP-10 with 75% sensitivity and 79% specificity, respectively. CONCLUSION: This study demonstrates that rESAT-6 and rCFP-10 can provide a sensitive and specific diagnosis of TB. In addition, it was shown that IL-2 may be serving as a marker for discriminating LTBI and active TB

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Guidelines for the use of interferon-gamma release assays in the diagnosis of tuberculosis infection / Guía de práctica clínica sobre el uso de las pruebas de liberación de interferón-gamma para el diagnóstico de infección tuberculosa

INTRODUCTION: Interferon-gamma release assays are widely used for the diagnosis of tuberculosis infection in low-prevalence countries. However, there is no consensus on their application. The objective of this study was to develop guidelines for the use of interferon-gamma release assays in specific clinical scenarios in Spain. METHODS: A panel of experts comprising specialists in infectious diseases, respiratory diseases, microbiology, pediatrics and preventive medicine, together with a methodologist, formulated the clinical questions and outcomes of interest. They conducted a systematic literature search, summarized the evidence and rated its quality, and prepared the recommendations following the GRADE (Grading of Recommendations of Assessment Development and Evaluations) methodology. RESULTS: The panel prepared recommendations on the use of interferon-gamma release assays for the diagnosis of tuberculosis infection in the contact-tracing study (both adults and children), health care workers, immunosuppressed patients (patients infected with human immunodeficiency virus, patients with chronic immunomediated inflammatory diseases due to start biological therapy and patients requiring organ transplant) and for the diagnosis of active tuberculosis. Most recommendations were weak, mainly due to the lack of good quality evidence to balance the clinical benefits and disadvantages of the interferon-gamma release assays as compared with the tuberculin skin test. CONCLUSION: This document provides evidence-based guidance on the use of interferon-gamma release assays for the diagnosis of tuberculosis infection in patients at risk of tuberculosis or with suspicion of active disease. The guidelines will be applicable in specialist and primary care and in public health settings

INTRODUCCIÓN: Las técnicas de detección in vitro de interferón-gamma (IGRA, del inglés interferon-gamma release assays) están ampliamente implantadas para el diagnóstico de infección tuberculosa en países de baja prevalencia. Sin embargo, no hay consenso sobre su aplicación. El objetivo fue desarrollar una guía de práctica clínica para el uso de los IGRA en los diferentes escenarios clínicos en España. MÉTODOS: Un grupo de expertos compuesto por especialistas en enfermedades infecciosas, enfermedades respiratorias, microbiología, pediatría y medicina preventiva, junto con un metodólogo formularon las preguntas clínicas y los desenlaces de interés, llevaron a cabo una búsqueda sistemática de la literatura, sintetizaron la evidencia y graduaron su calidad, y formularon las recomendaciones siguiendo la metodología Grading of Recommendations of Assessment Development and Evaluations(GRADE). RESULTADOS: El grupo de trabajo formuló las recomendaciones sobre el uso de los IGRA para el diagnóstico de infección tuberculosa en el estudio de contactos (adultos y niños), trabajadores sanitarios, pacientes inmunosuprimidos (pacientes infectados por el virus de la inmunodeficiencia humana, pacientes afectos de enfermedades inflamatorias inmunomediadas candidatos a terapias biológicas y pacientes que requieren trasplante de órganos), y en el diagnóstico de enfermedad tuberculosa activa. La mayor parte de las recomendaciones fueron débiles, principalmente debido a la falta de evidencia de calidad para establecer un balance entre beneficios y daños de los IGRA en comparación con la prueba de la tuberculina. CONCLUSIÓN: Este documento proporciona una guía basada en la evidencia para el uso de los IGRA en el diagnóstico de infección tuberculosa en pacientes en riesgo de tuberculosis o con sospecha de enfermedad activa. Esta guía es aplicable en la atención especializada y primaria, y salud pública

Executive summary of the guidelines for the use of interferon-gamma release assays in the diagnosis of tuberculosis infection / Sumario Ejecutivo de la Guía de Práctica Clínica sobre el uso de las pruebas de liberación de interferón-gamma para el diagnóstico de infección tuberculosa

Interferon-gamma release assays are widely used for the diagnosis of tuberculosis infection in Spain. However, there is no consensus on their application in specific clinical scenarios. To develop a guideline for their use, a panel of experts comprising specialists in infectious diseases, respiratory diseases, microbiology, pediatrics and preventive medicine, together with a methodologist, conducted a systematic literature search, summarized the findings, rated the quality of the evidence, and formulated recommendations following the GRADE (Grading of Recommendations of Assessment Development and Evaluations) methodology. This document provides evidence-based guidance on the use of interferon-gamma release assays for the diagnosis of tuberculosis infection in patients at the risk of tuberculosis or suspected of having active disease. The guidelines will be applicable to specialist and primary care, and public health

Las técnicas de detección in vitro de interferón-gamma (IGRA, del inglés interferon-gamma release assays), están ampliamente implantadas para el diagnóstico de infección tuberculosa en España. Sin embargo, no hay consenso sobre su aplicación en diferentes escenarios clínicos. Para desarrollar una guía de práctica clínica sobre su uso, un grupo de trabajo compuesto por especialistas en enfermedades infecciosas, neumología, microbiología, pediatría y medicina preventiva, junto con un metodólogo, llevaron a cabo una búsqueda sistemática de la literatura, sintetizaron la evidencia y gradaron su calidad, y formularon las recomendaciones siguiendo el método GRADE (Grading of Recommendations of Assessment Development and Evaluations). Este documento proporciona una guía basada en la evidencia para el uso de los IGRA para el diagnóstico de infección tuberculosa en pacientes en riesgo de tuberculosis o con sospecha de enfermedad activa. Esta guía es aplicable en la atención especializada y primaria, y salud pública

Impaired in-vitro responses to IL-12 and IFN-γ in Iranian patients with Mendelian susceptibility to mycobacterial disease

INTRODUCTION: Diagnosis of specific molecular defects of Mendelian susceptibility to mycobacterial diseases (MSMD) patients is important with respect to their clinical outcomes and their response to therapy. The aim of this study was to perform functional tests on blood samples of a group of patients who were suspected of having MSMD. METHODS: This study was performed on 11 cases who had mycobacterial infections and suspected MSMD. Whole blood cell culture was performed in presence of different stimulators. The supernatants were assayed for IFN-γ, IL-12p40 by ELISA method. RESULTS: All patients presented with complications of BCG vaccine in the form of localised lymphadenitis or disseminated BCG infection and chronic mycobacterial osteomyelitis. Infections with Salmonellaspecies occurred in two patients. In-vitro studies showed that 10 cases had impaired response to IL-12. However, the baseline levels of IL-12p40 were normal, while one of our patients may have a potential IFN-γ signalling defect or an IL-12p40 defect. CONCLUSIONS: Early detection of MSMD and commencing of appropriate combination therapy could prevent severe or even fatal complications of uncontrolled mycobacterial infections

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Peripheral blood mononuclear cells from severe asthmatic children release lower amounts of IL-12 and IL-4 after LPS stimulation

INTRODUCTION: Asthma is an inflammatory disorder of the airways associated with bronchial hyperresponsiveness, airway obstruction, and increased mucus production, with a predominance of type 2 immune response (Th2). According to the hygiene hypothesis, exposure to environmental bacterial lipopolysaccharide (LPS) may induce a type 1 immune response (Th1), modulating the development of asthma. OBJECTIVE: In this study we investigated cytokine production by peripheral blood mononuclear cells (PBMC) from children and adolescents with severe asthma, in response to LPS stimulation in vitro. MATERIALS AND METHODS: 26 children were selected: 13 severe asthmatics and 13 healthy controls, aged between 5 and 18 years. They were evaluated through routine medical history, physical examination and lung function test to diagnose severe asthma. Allergy status was confirmed by skin prick test and specific IgE assay. We collected blood samples to analyse in vitro LPS-induced cytokines release by PBMC. RESULTS: PBMC from severe asthmatic children produced lower levels of IL-12p70 in basal conditions and after 12 and 24 h stimulation with LPS compared to healthy controls. PBMC from severe asthmatic children produced lower levels of IL-4 after 24 h LPS stimulation compared to healthy controls. PBMC from severe asthmatic children produced more levels IL-17 and IL-10 after stimulus with LPS compared to healthy controls. The release of IFN-γ, IL-5 and TNF-α by PBMC from severe asthmatic children was similar to healthy controls. CONCLUSION: Our results demonstrate that LPS directly influence the cytokine profile of PBMC in children with severe asthma. These observations may be potentially helpful in developing new treatment strategies

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Expresión de interferón gamma en la infección por el virus del papiloma humano y por Chlamydia trachomatis en muestras cervicales / Expression of gamma interferon during HPV and Chlamydia trachomatis infection in cervical samples

INTRODUCCIÓN: El objetivo del presente trabajo fue medir la expresión de interferón gamma en la infección por VPH y por Chlamydia trachomatis en pacientes con lesión intraepitelial escamosa. MÉTODO: Se incluyeron 100 muestras de pacientes diagnosticadas por colposcopía, con y sin lesión intraepitelial escamosa, en quienes se efectuó el diagnóstico de infección por VPH y/o C. trachomatis. Se cuantificó la expresión relativa de interferón gamma con la prueba de transcriptasa reversa-PCR en tiempo real (RT-PCR). RESULTADOS: Las unidades relativas de la expresión de interferón gamma fueron de 13, 1,8 y 0,3 en la coinfección por VPH y C. trachomatis, en la infección por VPH y en la infección por C. trachomatis, respectivamente. CONCLUSIÓN: La infección por VPH y por C. trachomatis puede constituir un factor estimulante de la expresión de interferón gamma

INTRODUCTION: The aim of this study was to mesure the expression of gamma interferon in HPV and Chlamydia trachomatis infection in squamous intraepithelial lesions. METHOD: Samples from 100 patients diagnosed by colposcopy with or without squamous intraepithelial lesions were used in the present study. Each patient was found to be infected by HPV and C. trachomatis. Relative gamma interferon mRNA expression was assessed using a real-time reverse transcriptase PCR assay (RT-PCR). RESULTS: The relative units of expression of gamma interferon mRNA were 13, 1.8 and 0.3, for HPV and C. trachomatis co-infection, or HPV or C. trachomatis infection, respectively. CONCLUSIÓN: HPV and C. trachomatis could overstimulate the expression of gamma interferon