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BACKGROUND: Chronic airway inflammatory diseases, such as chronic rhinosinusitis with nasal polyps and asthma, show increased nasal Staphylococcus aureus colonization. Staphylococcus aureus-derived serine protease-like protein (Spl) D and other closely related proteases secreted by S aureus have recently been identified as inducers of allergic asthma in human subjects and mice, but their mechanism of action is largely unknown. OBJECTIVE: We investigated the role of recombinant SplD in driving TH2-biased responses and IgE formation in a murine model of allergic asthma. METHODS: Allergic asthma was induced in C57BL/6 J wild-type mice, Toll-like receptor (TLR) 4 knockout (Tlr4-/-) mice, and recombination-activating gene (Rag2) knockout (Rag2-/-) mice by means of repeated intratracheal applications of SplD. Inflammatory parameters in the airways were assessed by means of flow cytometry, ELISA, Luminex, and immunohistochemistry. Serum SplD-specific IgE levels were analyzed by using ELISA. RESULTS: We observed that repeated intratracheal exposure to SplD led to IL-33 and eotaxin production, eosinophilia, bronchial hyperreactivity, and goblet cell hyperplasia in the airways. Blocking IL-33 activity with a soluble ST2 receptor significantly decreased the numbers of eosinophils, IL-13+ type 2 innate lymphoid cells and IL-13+CD4+ T cells and IL-5 and IL-13 production by lymph node cells but had no effect on IgE production. SplD-induced airway inflammation and IgE production were largely dependent on the presence of the functional adaptive immune system and independent of TLR4 signaling. CONCLUSION: The S aureus-derived protein SplD is a potent allergen of S aureus and induces a TH2-biased inflammatory response in the airways in an IL-33-dependent but TRL4-independent manner. The soluble ST2 receptor could be an efficient strategy to interfere with SplD-induced TH2 inflammation but does not prevent the allergic sensitization.
Assuntos
Asma/imunologia , Proteínas de Bactérias/toxicidade , Proteína 1 Semelhante a Receptor de Interleucina-1/imunologia , Interleucina-33/imunologia , Serina Proteases/toxicidade , Staphylococcus aureus/imunologia , Animais , Asma/induzido quimicamente , Asma/genética , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/patologia , Modelos Animais de Doenças , Proteína 1 Semelhante a Receptor de Interleucina-1/genética , Interleucina-33/genética , Camundongos , Camundongos Knockout , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Staphylococcus aureus/patogenicidade , Células Th2/imunologia , Células Th2/patologiaRESUMO
BACKGROUND: Clinical and experimental studies have identified a crucial role for IL-33 and its receptor ST2 in allergic asthma. Inhalation of traffic-related pollutants, such as diesel exhaust particles (DEP), facilitates the development of asthma and can cause exacerbations of asthma. However, it is unknown whether IL-33/ST2 signalling contributes to the enhancing effects of air pollutants on allergic airway responses. OBJECTIVE: We aim to investigate the functional role of IL-33/ST2 signalling in DEP-enhanced allergic airway responses, using an established murine model. METHODS: C57BL/6J mice were exposed to saline, DEP alone, house dust mite (HDM) alone or combined DEP+HDM. To inhibit IL-33 signalling, recombinant soluble ST2 (r-sST2) was given prophylactically (ie, during the whole experimental protocol) or therapeutically (ie, at the end of the experimental protocol). Airway hyperresponsiveness and the airway inflammatory responses were assessed in bronchoalveolar lavage fluid (BALF) and lung. RESULTS: Combined exposure to DEP+HDM increased IL-33 and ST2 expression in lung, elevated inflammatory responses and bronchial hyperresponsiveness compared to saline, sole DEP or sole HDM exposure. Prophylactic interference with the IL-33/ST2 signalling pathway impaired the DEP-enhanced allergic airway inflammation in the BALF, whereas effects on lung inflammation and airway hyperresponsiveness were minimal. Treatment with r-sST2 at the end of the experimental protocol did not modulate the DEP-enhanced allergic airway responses. CONCLUSION: Our data suggest that the IL-33/ST2 pathway contributes to the onset of DEP-enhanced allergic airway inflammation.
Assuntos
Poluentes Atmosféricos/efeitos adversos , Interleucina-33/metabolismo , Hipersensibilidade Respiratória/etiologia , Hipersensibilidade Respiratória/metabolismo , Transdução de Sinais , Alérgenos/imunologia , Animais , Biomarcadores , Modelos Animais de Doenças , Feminino , Proteína 1 Semelhante a Receptor de Interleucina-1/metabolismo , Leucócitos/imunologia , Leucócitos/metabolismo , Linfócitos/imunologia , Linfócitos/metabolismo , Camundongos , Material Particulado/efeitos adversos , Pyroglyphidae/imunologia , Proteínas Recombinantes/farmacologia , Hipersensibilidade Respiratória/tratamento farmacológico , Hipersensibilidade Respiratória/patologia , Mucosa Respiratória/imunologia , Mucosa Respiratória/metabolismoRESUMO
BACKGROUND: Vaccination against severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) was one crucial element to overcome the coronavirus disease 2019 (COVID-19) pandemic. Even though anaphylaxis to vaccines is rare, 47 patients came to the Allergy Unit at the University Hospital Graz, Austria, reporting immediate anaphylactoid symptoms after administration of COVID-19 vaccines. In addition, 29 patients with known drug-induced anaphylaxis wanted to be tested for a possible sensitization against COVID-19 vaccines or excipients, such as polyethylene glycol (PEG) or polysorbate 80 (PS80) before the first COVID-19 vaccination. Skin prick tests and intradermal tests were performed in all 76 patients, mostly using PEG 2000, and/or PS80. Skin prick tests with COVID-19 vaccines were performed depending on availability. OBJECTIVE: Our aim was to characterize this patient cohort in terms of patients' anaphylactoid responses, their willingness to future vaccinations against SARS-Cov2, and reasons for their decision. METHODS: We developed a questionnaire and analyzed 34 completed copies. RESULTS: Of the 47 patients with anaphylactoid reactions to COVID-19 vaccination, most were female (40 female/7 male). The skin tests, even when performed with the respective COVID-19 vaccine, were negative in all but one patient. Most patients who experienced anaphylactoid reactions after a COVID-19 vaccination, did not want another COVID-19 vaccination at the time of answering the questionnaire because of anxiety for another anaphylactoid response at the next shot. Premedication with antihistamines significantly lowered (nâ¯= 74 vaccinations) the severity of anaphylactoid responses after COVID-19 vaccinations. CONCLUSION: Anxiety about another anaphylactoid episode hinders patients to be vaccinated against SARS-CoV2 again. Premedication with antihistamines and collaboration of allergologists with psychologists might lower the risk of an anaphylactic/anaphylactoid response as well anxiety in drug-induced anaphylactic patients.
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Leukemic cutaneous T-cell lymphomas (L-CTCL) are lymphoproliferative disorders of skin-homing mature T-cells causing severe symptoms and high mortality through chronic inflammation, tissue destruction, and serious infections. Despite numerous genomic sequencing efforts, recurrent driver mutations have not been identified, but chromosomal losses and gains are frequent and dominant. We integrated genomic landscape analyses with innovative pharmacologic interference studies to identify key vulnerable nodes in L-CTCL. We detected copy number gains of loci containing the STAT3/5 oncogenes in 74% (n = 17/23) of L-CTCL, which correlated with the increased clonal T-cell count in the blood. Dual inhibition of STAT3/5 using small-molecule degraders and multi-kinase blockers abolished L-CTCL cell growth in vitro and ex vivo, whereby PAK kinase inhibition was specifically selective for L-CTCL patient cells carrying STAT3/5 gains. Importantly, the PAK inhibitor FRAx597 demonstrated encouraging anti-leukemic activity in vivo by inhibiting tumor growth and disease dissemination in intradermally xenografted mice. We conclude that STAT3/5 and PAK kinase interaction represents a new therapeutic node to be further explored in L-CTCL.
Assuntos
Linfoma Cutâneo de Células T , Quinases Ativadas por p21 , Animais , Camundongos , Genômica , Xenoenxertos , Linfoma Cutâneo de Células T/tratamento farmacológicoRESUMO
Staphylococcus aureus (S. aureus) can secrete a broad range of virulence factors, among which staphylococcal serine protease-like proteins (Spls) have been identified as bacterial allergens. The S. aureus allergen serine protease-like protein D (SplD) induces allergic asthma in C57BL/6J mice through the IL-33/ST2 signaling axis. Analysis of C57BL/6J, C57BL/6N, CBA, DBA/2, and BALB/c mice treated with intratracheal applications of SplD allowed us to identify a frameshift mutation in the serine (or cysteine) peptidase inhibitor, clade A, and member 3I (Serpina3i) causing a truncated form of SERPINA3I in BALB/c, CBA, and DBA/2 mice. IL-33 is a key mediator of SplD-induced immunity and can be processed by proteases leading to its activation or degradation. Full-length SERPINA3I inhibits IL-33 degradation in vivo in the lungs of SplD-treated BALB/c mice and in vitro by direct inhibition of mMCP-4. Collectively, our results establish SERPINA3I as a regulator of IL-33 in the lungs following exposure to the bacterial allergen SplD, and that the asthma phenotypes of mouse strains may be strongly influenced by the observed frameshift mutation in Serpina3i. The analysis of this protease-serpin interaction network might help to identify predictive biomarkers for type-2 biased airway disease in individuals colonized by S. aureus.
Assuntos
Alérgenos/imunologia , Proteínas de Bactérias/imunologia , Interleucina-33/imunologia , Serina Proteases/imunologia , Infecções Estafilocócicas/imunologia , Staphylococcus aureus/imunologia , Animais , Asma/imunologia , Feminino , Mutação da Fase de Leitura/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos CBA , Camundongos Endogâmicos DBA , Peptídeo Hidrolases/imunologia , Serina Endopeptidases/imunologia , Serpinas/imunologiaRESUMO
Staphylococcus aureus persistently colonizes the nostrils of one-third of the population but colonizes the sinus mucosa in up to 90% of patients with nasal polyps, implying a possible role in airway disease. Recent findings give new mechanistic insights into the ability of S. aureus to trigger type 2 inflammatory responses in the upper and lower airways. This novel concept of a S. aureus-driven chronic airway inflammatory disease suggests a new understanding of disease triggers. This article reviews the role of S. aureus in chronic inflammatory airway diseases and discusses possible therapeutic approaches to target S. aureus.
Assuntos
Infecções Respiratórias/microbiologia , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/fisiologia , Asma/complicações , Asma/epidemiologia , Asma/etiologia , Bacteriófagos/fisiologia , Suscetibilidade a Doenças/imunologia , Humanos , Imunização , Imunoglobulina E/imunologia , Fagocitose/imunologia , Mucosa Respiratória/imunologia , Mucosa Respiratória/metabolismo , Mucosa Respiratória/microbiologia , Infecções Respiratórias/diagnóstico , Infecções Respiratórias/tratamento farmacológico , Infecções Respiratórias/imunologia , Infecções Estafilocócicas/diagnóstico , Infecções Estafilocócicas/tratamento farmacológico , Infecções Estafilocócicas/imunologia , Staphylococcus aureus/virologia , Nanomedicina Teranóstica , VirulênciaRESUMO
UNLABELLED: The ability of flavoenzymes to reduce dioxygen varies greatly, and is controlled by the protein environment, which may cause either a rapid reaction (oxidases) or a sluggish reaction (dehydrogenases). Previously, a 'gatekeeper' amino acid residue was identified that controls the reactivity to dioxygen in proteins from the vanillyl alcohol oxidase superfamily of flavoenzymes. We have identified an alternative gatekeeper residue that similarly controls dioxygen reactivity in the grass pollen allergen Phl p 4, a member of this superfamily that has glucose dehydrogenase activity and the highest redox potential measured in a flavoenzyme. A substitution at the alternative gatekeeper site (I153V) transformed the enzyme into an efficient oxidase by increasing dioxygen reactivity by a factor of 60,000. An inverse exchange (V169I) in the structurally related berberine bridge enzyme (BBE) decreased its dioxygen reactivity by a factor of 500. Structural and biochemical characterization of these and additional variants showed that our model enzymes possess a cavity that binds an anion and resembles the 'oxyanion hole' in the proximity of the flavin ring. We showed also that steric control of access to this site is the most important parameter affecting dioxygen reactivity in BBE-like enzymes. Analysis of flavin-dependent oxidases from other superfamilies revealed similar structural features, suggesting that dioxygen reactivity may be governed by a common mechanistic principle. DATABASE: Structural data are available in PDB database under the accession numbers 4PVE, 4PVH, 4PVJ, 4PVK, 4PWB, 4PWC and 4PZF.