Sudan red dye: a new agent causing type-2 occupational asthma

ackground: Sudan red or 1‐[(2‐methoxyphenyl)azo]‐2‐naphthol is a low molecular weight azoic agent widely used in industry, particularly in the production of hair dyes. The use of this product in the food industry is prohibited due to its potential carcinogenic effect, but no respiratory involvement has been reported to date. Case presentation: We present the case of a 46‐year‐old female patient who had been working in a cosmetics packaging company for 20 years. The patient developed occupational asthma to a red azo dye known as Sudan red. The diagnosis was confirmed by specific bronchial provocation test. Induced sputum samples were obtained previously and in the 24 h following the procedure, with a rise in the percentage of eosinophils from 10 to 65%. Conclusions: This report describes the case of a patient who developed OA caused by exposure to an azoic dye called Sudan red. The clinical and analytical features suggest a type 2‐related asthma; however, we are not yet able to confirm the specific pathophysiological mechanism. The extensive use of azo dyes in industry means that it is particularly important to describe their implications for health, which are probably underestimated at present

Mycobacterium manresensis induces trained immunity in vitro

The COVID-19 pandemic posed a global health crisis, with new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants weakening vaccine-driven protection. Trained immunity could help tackle COVID-19 disease. Our objective was to analyze whether heat-killed Mycobacterium manresensis (hkMm), an environmental mycobacterium, induces trained immunity and confers protection against SARS-CoV-2 infection. To this end, THP-1 cells and primary monocytes were trained with hkMm. The increased secretion of tumor necrosis factor alpha (TNF-α), interleukin (IL)-6, IL-1β, and IL-10, metabolic activity, and changes in epigenetic marks suggested hkMm-induced trained immunity in vitro. Healthcare workers at risk of SARS-CoV-2 infection were enrolled into the MANRECOVID19 clinical trial (NCT04452773) and were administered Nyaditum resae (NR, containing hkMm) or placebo. No significant differences in monocyte inflammatory responses or the incidence of SARS-CoV-2 infection were found between the groups, although NR modified the profile of circulating immune cell populations. Our results show that M. manresensis induces trained immunity in vitro but not in vivo when orally administered as NR daily for 14 days. Biological sciences; Molecular biology; Immunology; Microbiolog

Mycobacterium manresensis induces trained immunity in vitro

The COVID-19 pandemic posed a global health crisis, with new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants weakening vaccine-driven protection. Trained immunity could help tackle COVID-19 disease. Our objective was to analyze whether heat-killed Mycobacterium manresensis (hkMm), an environmental mycobacterium, induces trained immunity and confers protection against SARS-CoV-2 infection. To this end, THP-1 cells and primary monocytes were trained with hkMm. The increased secretion of tumor necrosis factor alpha (TNF-α), interleukin (IL)-6, IL-1β, and IL-10, metabolic activity, and changes in epigenetic marks suggested hkMm-induced trained immunity in vitro. Healthcare workers at risk of SARS-CoV-2 infection were enrolled into the MANRECOVID19 clinical trial (NCT04452773) and were administered Nyaditum resae (NR, containing hkMm) or placebo. No significant differences in monocyte inflammatory responses or the incidence of SARS-CoV-2 infection were found between the groups, although NR modified the profile of circulating immune cell populations. Our results show that M. manresensis induces trained immunity in vitro but not in vivo when orally administered as NR daily for 14 days.The MANRECOVID19 clinical trial has been sponsored by the Reig Jofre Group. This research was funded by the Consorcio Centro de Investigación Biomédica en Red (CIBERES and CIBEREHD) and the European Union’s Horizon 2020 research and innovation programme under grant agreement No 847762. MDH is supported by a Margarita Salas grant from NextGenerationEU. LS-M is supported by Juan de la Cierva fellowship (FJC2019-041213-I). NI-U is supported by the Spanish Ministry of Science and Innovation (grant PID2020-117145RB-I00), EU HORIZON-HLTH-2021-CORONA-01 (grant 101046118), and institutional funding from Grifols, Pharma Mar, HIPRA, Amassence, and Palobiofarma. The Innate Immunity lab and the UTE are accredited by the Catalan Agency for Management of University and Research Grants (2017-SGR-490/2021-SGR-01186, 2021-SGR-00931, and 2017-SGR-500/2021 SGR 00920). IGTP is a member of the CERCA network of institutes supported by the Health Department of the Government of Catalonia.info:eu-repo/semantics/publishedVersio

Effects of diesel exhaust particles on allergic and chemical-induced asthma

La contaminació ambiental indueix i exacerba l'asma, encara que els mecanismes immunològics pels quals les partícules dièsel (DEP) modulen l'asma no estan del tot descrits i semblen diferir segons l'endotip asmàtic: tipus 2 (T2), subdividit segons si la resposta és dirigida per les cèl·lules Th2 o ILC2; el no T2; i l'endotip mixt Th2/Th17. S'ha vist que en l'endotip T2 la contaminació ambiental incrementa la sensibilització a certs al·lèrgens ambientals, mentre que les DEPS actuen com a adjuvant en la inducció de l'asma al·lèrgic. En l'endotip T2 Th2-mediat, les DEPS podrien augmentar la capacitat antigènica de certs aeroal·lèrgens; mentre que en l'endotip ILC2-mediat, les DEPS podrien estimular l'epiteli pulmonar a produir alts nivells d'interleucina (IL)-33, IL-25 i la limfopoietina tímica estromal (TLSP), citocines relacionades amb la via de les ILC2 i l'asma eosinofílica. A l'endotip no T2, les DEPS podrien agreujar l'asma neutrofílica augmentant la producció de citocines com la IL-17A i IL-17F. Finalment, s'ha descrit que la coexposició a al·lergen de closca de soja (SHE) i DEPS podria afavorir una resposta mixta Th2/Th17, involucrant no només la inflamació Th2 amb IL-4, IL-5, IL-13 i IL 10, sinó també una resposta Th17 amb nivells creixents de quimiocines com ara la IL-17A, IL-17F i CCL20. La present tesi doctoral té com objectiu estudiar l'efecte de l'exposició a les DEPS sobre dos endotips d'asma diferents, mitjançant l'avaluació de la funció respiratòria, la hiperreactivitat bronquial (AHR), la inflamació pulmonar, l'estrès oxidatiu i el patró de deposició de les DEPS en les vies aèries. El primer estudi avalua els efectes de l'exposició a DEPS en un model murí d'asma químic a sals d'amoni de persulfat (AP), un endotip no T2. Aquest estudi demostra que la sensibilització i la inhalació d'AP indueix un endotip asmàtic tipus T2 baix, caracteritzat per funció pulmonar asmàtica, elevats nivells de resistència de les vies aèries (Rn), elastància tissular i AHR; i inflamació de les vies respiratòries amb eosinofília i baixos nivells de cèl·lules dendrítiques (CD) tolerogèniques. El grup exposat a les DEPS es caracteritza per tenir una funció pulmonar alterada; inflamació de les vies respiratòries amb neutrofília, nivells superiors de CDs relacionades amb la via Th2 nivells inferiors de macròfags alveolars; i nivells reduïts de les citocines IL-13 i IFN-γ. El grup coexposat a AP i DEPS presenta signes d'asma exacerbat, consistent principalment en una funció pulmonar típicament asmàtica; inflamació de les vies respiratòries amb nivells superiors de CDs sensibles a l'estrès oxidatiu i quantitats més baixes de macròfags totals; presentant també una deposició diferencial de les DEPS en els pulmons, trobant-se principalment en vies superiors. El segon estudi analitza els efectes de l'exposició a DEPS en un model murí exposat a baixes concentracions de SHE, un endotip T2. En aquest segon capítol, la inhalació de SHE indueix una lleu inflamació de les vies respiratòries amb nivells elevats d'eosinòfils, cèl·lules B, CDs relacionades amb la via Th2, monòcits totals i residents, així com nivells reduïts de NKS, CDs tolerogèniques, macròfags totals i alveolars. La inhalació de les DEPS desencadena neutrofília a les vies respiratòries. Finalment, la inhalació de SHE i DEPS conjuntament indueix un endotip d'asma mixt Th2/Th17, caracteritzat per una funció pulmonar asmàtica amb nivells elevats de AHR i Rn; inflamació de les vies respiratòries amb nivells superiors d'immunoglobulina (Ig) de tipus E específica a SHE en sèrum, H2O2 en rentat broncoalveolar (BAL), NKS, CDs sensibles a l'estrès oxidatiu i nivells més baixos de CDs relacionades amb la via Th1.La contaminación ambiental induce y exacerba el asma, aunque los mecanismos inmunológicos por los cuales las partículas diésel (DEP) modulan el asma no están del todo descritos y parecen diferir según el endotipo asmático: tipo 2 (T2), subdividido según si la respuesta es dirigida por las células Th2 o ILC2; el no T2; y el endotipo mixto Th2/Th17. Se ha visto que en el endotipo T2 la contaminación ambiental incrementa la sensibilización a ciertos alérgenos ambientales, mientras que las DEPs actúan como adyuvante en la inducción del asma alérgico. En el endotipo T2 Th2-mediado, las DEPs podrían aumentar la capacidad antigénica de ciertos aeroalérgenos; mientras que en el endotipo ILC2-mediado, las DEPs podrían estimular el epitelio pulmonar a producir altos niveles de interleucina (IL)-33, IL-25 y la limfopoietina tímica estromal (TLSP), citocinas relacionadas con la vía de las ILC2 y el asma eosinofílica. En el endotipo no T2, las DEPs podrían agravar el asma neutrofílica aumentando la producción de citocinas como la IL-17A e IL-17F. Por último, se ha descrito que la coexposición a alérgeno de cáscara de soja (SHE) y DEPs podría favorecer una respuesta mixta Th2/Th17, involucrando no solo la inflamación Th2 con IL-4, IL-5, IL-13 e IL-10, sino también una respuesta Th17 con niveles crecientes de quimiocinas tales como la IL-17A, IL-17F y CCL20. La presente tesis doctoral tiene como objetivo estudiar el efecto de la exposición a las DEPs sobre dos endotipos de asma diferentes, mediante la evaluación de la función respiratoria, la hiperreactividad bronquial (AHR), la inflamación pulmonar, el estrés oxidativo y el patrón de deposición de las DEPs en las vías aéreas. El primer estudio evalúa los efectos de la exposición a DEPs en un modelo murino de asma químico a sales de amonio de persulfato (AP), un endotipo no T2. Dicho estudio demuestra que la sensibilización y la inhalación de AP induce un endotipo asmático tipo T2 bajo, caracterizado por función pulmonar asmática, elevados niveles de resistencia de las vías aéreas (Rn), elastancia tisular y AHR; e inflamación de las vías respiratorias con eosinofilia y bajos niveles de células dendríticas (CD) tolerogénicas. El grupo expuesto a las DEPs se caracteriza por tener una función pulmonar alterada; inflamación de las vías respiratorias con neutrofilia, niveles superiores de CDs relacionadas con la vía Th2 niveles inferiores de macrófagos alveolares; y niveles reducidos de las citocinas IL-13 e IFN-γ. El grupo coexpuesto a AP y DEPs presenta signos de asma exacerbado, consistente principalmente en una función pulmonar típicamente asmática; inflamación de las vías respiratorias con niveles superiores de CDs sensibles al estrés oxidativo y cantidades más bajas de macrófagos totales; presentando también una deposición diferencial de las DEPs en los pulmones, hallándose principalmente en vías superiores. El segundo estudio analiza los efectos de la exposición a DEPs en un modelo murino expuesto a bajas concentraciones de SHE, un endotipo T2. En este segundo capítulo, la inhalación de SHE induce una leve inflamación de las vías respiratorias con niveles elevados de eosinófilos, células B, CDs relacionadas con la vía Th2, monocitos totales y residentes, así como niveles reducidos de NKs, CDs tolerogénicas, macrófagos totales y alveolares. La inhalación de las DEPs desencadena neutrofilia en las vías respiratorias. Por último, la inhalación de SHE y DEPs conjuntamente induce un endotipo de asma mixto Th2/Th17, caracterizado por una función pulmonar asmática con niveles elevados de AHR y Rn; inflamación de las vías respiratorias con niveles superiores de inmunoglobulina (Ig) de tipo E específica a SHE en suero, H2O2 en lavado broncoalveolar (BAL), NKs, CDs sensibles al estrés oxidativo y niveles más bajos de CDs relacionadas con la vía Th1.Air pollution is known to induce and exacerbate asthma pathophysiology, but the immunological effects of diesel exhaust particles (DEP) are still not well characterized and seem to differ depending on the pre-existing asthma endotype: type 2 (T2), which is subdivided further into Th2 and ILC2-driven; non-T2; and a mixed Th2/Th17 endotype. In the T2 endotype, traffic-related air pollution (TRAP) has been demonstrated to enhance respiratory sensitization to environmental allergens, while it has been suggested that DEPs act as adjuvants in the induction of allergic asthma. In the Th2-driven T2 endotype it seems that DEPs may increase the antigenic capacity of certain aeroallergens; while in the ILC2-driven T2 endotype, they may stimulate lung epithelial cells to produce high levels of interleukin (IL)-33, IL-25, and thymic stromal lymphopoietin (TSLP), cytokines related to ILC2s and eosinophilic asthma. In the non-T2 endotype, DEPs may aggravate neutrophilic asthma by increasing the production of both IL-17A and IL-17F cytokines. Lastly, our group reported that coexposure to soybean hull extract (SHE) and DEPs may favour a mixed Th17/Th2 response, not only involving a Th2 inflammation with IL-4, IL-5, IL-13 and IL-10, but also increasing levels of chemokines related to the Th17 response such as IL-17A, IL-17F, and CCL20 cytokines. The present doctoral thesis aims to study the effects of DEP exposure on two different asthma endotypes by assessing respiratory mechanics, airway hyperresponsiveness (AHR), innate and adaptive immune responses, oxidative stress and particle deposition patterns. The first study (Chapter 1) aims to study the effects of DEP exposure in a mouse model of chemical-induced asthma due to ammonium persulfate (AP), a non-type 2 endotype. This study demonstrates that sensitization and inhalation of AP induce a T2 low asthma endotype, characterized by asthma-like lung parameters such as higher levels of central airway resistance, tissue elastance, and AHR, and airway inflammation consisting in eosinophilia, and lower levels of tolerogenic dendritic cells (DC). The DEP-exposed group showed slightly modified lung mechanics, airway inflammation with neutrophilia, higher levels of Th2-related DCs, lower levels of alveolar macrophages, and reduced levels of cytokines such as IL-13 and IFN-γ. The group coexposed to AP and DEPs presented exacerbated signs of asthma consisting principally of asthma-like lung function, airway inflammation with higher levels of oxidative stress-sensitive DCs, lower total macrophages, and a differential pattern of DEPs deposition in the lungs, restricted to large conducting airways. The second study (Chapter 2) aims to study the effects of DEPs exposure in a mouse model exposed to non-asthmagenic doses of SHE, a Type 2 endotype. In this second chapter, the inhalation of 3 mg·ml-1 SHE induced a mild airway inflammation consisting in increased levels of eosinophils, B cells, Th2-related DCs, total and resident monocytes, and reductions in levels of NK cells, tolerogenic DCs, total and alveolar macrophages. The inhalation of DEPs alone triggers airway inflammation with neutrophilia. Lastly, the inhalation of SHE and DEPs together induced a mixed Th2/Th17 asthma endotype characterized by asthma-like lung function with higher levels of AHR and central airway resistance (Rn), airway inflammation consisting in higher levels of SHE-specific immunoglobulin (Ig)-E in serum, H2O2 in bronchoalveolar lavage (BAL), NK cells, oxidative stress-sensitive DCs, and lower levels of Th1-related DCs.Universitat Autònoma de Barcelona. Programa de Doctorat en Medicin

Biomarker Profiles Associated with COVID-19 Severity and Mortality

Introduction: The aim of this study was to analyze biomarkers that might predict the severity and progression of the SARS-CoV-2 infection, both in the acute phase and after recovery. Methods: Unvaccinated patients infected with the original strain of COVID-19 requiring ward (Group 1, n = 48) or ICU (Group 2, n = 41) admission were included. At the time of admission (visit 1), a clinical history was acquired, and blood samples were obtained. One and six months after discharge from the hospital (visits 2 and 3, respectively), a clinical history, lung function tests, and blood samples were carried out. At visit 2, patients also underwent a chest CT scan. Different cytokines (IL-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12p70, IL-13, IL-17A, G-CSF, GM-CSF, IFN-ɣ, MCP-1, MIP-1β, and TNF-α) and lung fibrosis biomarkers (YKL-40 and KL-6) were measured in blood samples obtained at visits 1, 2, and 3. Results: At visit 1, IL-4, IL-5, and IL-6 levels were higher in Group 2 (p = 0.039, 0.011, and 0.045, respectively), and IL-17 and IL-8 levels were higher in Group 1 (p = 0.026 and 0.001, respectively). The number of patients in Groups 1 and 2 who died during hospitalization was 8 and 11, respectively. YKL-40 and KL-6 levels were higher in patients who died. Serum YKL-40 and KL-6 levels determined at visit 2 correlated negatively with FVC (p = 0.022 and p = 0.024, respectively) and FEV1 (p = 0.012 and p = 0.032, respectively) measured at visit 3. KL-6 levels also correlated negatively with the diffusing capacity of the lungs for carbon monoxide (DLCO, p = 0.001). Conclusions: Patients who required ICU admission had higher levels of Th2 cytokines, while patients admitted to the ward showed an innate immune response activation, with IL-8 release and Th1/Th17 lymphocyte contribution. Increased levels of YKL-40 and KL-6 were associated with mortality in COVID-19 patients

Effects of diesel exhaust particle exposure on a murine model of asthma due to soybean.

Exposure to soybean allergens has been linked to asthma outbreaks. Exposure to diesel exhaust particles (DEP) has been associated with an increase in the risk of asthma and asthma exacerbation; however, in both cases the underlying mechanisms remain poorly understood, as does the possible interaction between the two entities.To investigate how the combination of soybean allergens and DEP can affect the induction or exacerbation of asthma in a murine model.BALB/c mice received intranasal instillations of saline, 3 or 5 mg protein/ml soybean hull extract (SHE), or a combination of one of these three solutions with DEP. Airway hyperresponsiveness (AHR), pulmonary inflammation in bronchoalveolar lavage, total serum immunoglobulin E and histological studies were assessed.A 5 mg protein/ml SHE solution was able by itself to enhance AHR (p = 0.0033), increase eosinophilic inflammation (p = 0.0003), increase levels of IL-4, IL-5, IL-13, IL-17A, IL-17F and CCL20, and reduce levels of IFN-γ. The combination of 5 mg protein/ml SHE with DEP also produced an increase in AHR and eosinophilic inflammation, but presented a slightly different cytokine profile with higher levels of Th17-related cytokines. However, while the 3 mg protein/ml SHE solution did not induce asthma, co-exposure with DEP resulted in a markedly enhanced AHR (p = 0.002) and eosinophilic inflammation (p = 0.004), with increased levels of IL-5, IL-17F and CCL20 and decreased levels of IFN-γ.The combination of soybean allergens and DEP is capable of triggering an asthmatic response through a Th17-related mechanism when the soybean allergen concentration is too low to promote a response by itself. DEP monitoring may be a useful addition to allergen monitoring in order to prevent new asthma outbreaks

Immunopathological Mechanisms of Bird-Related Hypersensitivity Pneumonitis

Bird-related hypersensitivity pneumonitis (BRHP) is an interstitial lung disease induced by avian proteins. The immunopathological pathways involved in the disease are still unknown. This study assesses the cellular immune response and the cytokine pattern in a mouse model of BRHP. On days −3 and −1, mice were intraperitoneally sensitized with commercial pigeon serum (PS) or saline. Intranasal instillations with PS or saline were carried out on three consecutive days/week over either 3 weeks (Group 1) or 12 weeks (Group 2). Leukocyte and cytokine patterns in lung tissue and pulmonary inflammation in bronchoalveolar lavage (BAL) were analysed. Both groups presented increases in resident monocytes, interstitial macrophages and type 2 dendritic cells (DCs), but also reductions in inflammatory monocytes, alveolar macrophages and tolerogenic DCs compared with their control groups. Group 1 had increased levels of eosinophils and T cells with reductions in neutrophils and B cells, while Group 2 showed high levels of B cells. Both groups exhibited increases in Th1 and Th2 cytokines. Group 2 also showed increased levels of IL-23, a Th17 cytokine. Increased levels of neutrophils, eosinophils and lymphocytes were observed in BAL samples of both groups compared with controls. In the first stages of BRHP, there is a mixed Th1/Th2 immune response, while during the progression of the disease, although there is a Th1 response, the cytokine levels seem to indicate a switch towards a Th2/Th17 mixed response

Immunopathological Mechanisms of Bird-Related Hypersensitivity Pneumonitis

Bird-related hypersensitivity pneumonitis (BRHP) is an interstitial lung disease induced by avian proteins. The immunopathological pathways involved in the disease are still unknown. This study assesses the cellular immune response and the cytokine pattern in a mouse model of BRHP. On days −3 and −1, mice were intraperitoneally sensitized with commercial pigeon serum (PS) or saline. Intranasal instillations with PS or saline were carried out on three consecutive days/week over either 3 weeks (Group 1) or 12 weeks (Group 2). Leukocyte and cytokine patterns in lung tissue and pulmonary inflammation in bronchoalveolar lavage (BAL) were analysed. Both groups presented increases in resident monocytes, interstitial macrophages and type 2 dendritic cells (DCs), but also reductions in inflammatory monocytes, alveolar macrophages and tolerogenic DCs compared with their control groups. Group 1 had increased levels of eosinophils and T cells with reductions in neutrophils and B cells, while Group 2 showed high levels of B cells. Both groups exhibited increases in Th1 and Th2 cytokines. Group 2 also showed increased levels of IL-23, a Th17 cytokine. Increased levels of neutrophils, eosinophils and lymphocytes were observed in BAL samples of both groups compared with controls. In the first stages of BRHP, there is a mixed Th1/Th2 immune response, while during the progression of the disease, although there is a Th1 response, the cytokine levels seem to indicate a switch towards a Th2/Th17 mixed response