RESUMEN
BACKGROUND: Forkhead box protein 3 (FOXP3) is the master transcription factor in CD4+CD25hiCD127lo regulatory T (Treg) cells. Mutations in FOXP3 result in IPEX (immune dysregulation, polyendocrinopathy, enteropathy, X-linked) syndrome. Clinical presentation of IPEX syndrome is broader than initially described, challenging the understanding of the disease, its evolution, and treatment choice. OBJECTIVE: We sought to study the type and extent of immunologic abnormalities that remain ill-defined in IPEX, across genetic and clinical heterogeneity. METHODS: We performed Treg-cell-specific epigenetic quantification and immunologic characterization of severe "typical" (n = 6) and "atypical" or asymptomatic (n = 9) patients with IPEX. RESULTS: Increased number of cells with Treg-cell-Specific Demethylated Region demethylation in FOXP3 is a consistent feature in patients with IPEX, with (1) highest values in those with typical IPEX, (2) increased values in subjects with pathogenic FOXP3 but still no symptoms, and (3) gradual increase over the course of disease progression. Large-scale profiling using Luminex identified plasma inflammatory signature of macrophage activation and TH2 polarization, with cytokines previously not associated with IPEX pathology, including CCL22, CCL17, CCL15, and IL-13, and the inflammatory markers TNF-α, IL-1A, IL-8, sFasL, and CXCL9. Similarly, both Treg-cell and Teff compartments, studied by Mass Cytometry by Time-Of-Flight, were skewed toward the TH2 compartment, especially in typical IPEX. CONCLUSIONS: Elevated TSDR-demethylated cells, combined with elevation of plasmatic and cellular markers of a polarized type 2 inflammatory immune response, extends our understanding of IPEX diagnosis and heterogeneity.
Asunto(s)
Enfermedades Genéticas Ligadas al Cromosoma X , Poliendocrinopatías Autoinmunes , Humanos , Factores de Transcripción Forkhead , Linfocitos T Reguladores , Mutación , Epigénesis GenéticaRESUMEN
BACKGROUND: Data on the safety and efficacy of coronavirus disease 2019 (COVID-19) vaccination in people with a range of primary immunodeficiencies (PIDs) are lacking because these patients were excluded from COVID-19 vaccine trials. This information may help in clinical management of this vulnerable patient group. OBJECTIVE: We assessed humoral and T-cell immune responses after 2 doses of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) messenger RNA (mRNA) vaccines in patients with PID and functional B-cell defects. METHODS: A double-center retrospective review was performed of patients with PID who completed COVID-19 mRNA vaccination and who had humoral responses assessed through SARS-CoV-2 spike protein receptor binding domain (RBD) IgG antibody levels with reflex assessment of the antibody to block RBD binding to angiotensin-converting enzyme 2 (ACE2; hereafter referred to as ACE2 receptor blocking activity, as a surrogate test for neutralization) and T-cell response evaluated by an IFN-γ release assay. Immunization reactogenicity was also reviewed. RESULTS: A total of 33 patients with humoral defect were evaluated; 69.6% received BNT162b2 vaccine (Pfizer-BioNTech) and 30.3% received mRNA-1273 (Moderna). The mRNA vaccines were generally well tolerated without severe reactions. The IFN-γ release assay result was positive in 24 (77.4%) of 31 patients. Sixteen of 33 subjects had detectable RBD-specific IgG responses, but only 2 of these 16 subjects had an ACE2 receptor blocking activity level of ≥50%. CONCLUSION: Vaccination of this cohort of patients with PID with COVID-19 mRNA vaccines was safe, and cellular immunity was stimulated in most subjects. However, antibody responses to the spike protein RBD were less consistent, and, when detected, were not effective at ACE2 blocking.
Asunto(s)
Vacuna nCoV-2019 mRNA-1273/inmunología , Vacuna BNT162/inmunología , COVID-19/inmunología , COVID-19/prevención & control , Enfermedades de Inmunodeficiencia Primaria/inmunología , Vacuna nCoV-2019 mRNA-1273/administración & dosificación , Vacuna nCoV-2019 mRNA-1273/efectos adversos , Adulto , Anciano , Anticuerpos Antivirales/biosíntesis , Linfocitos B/inmunología , Vacuna BNT162/administración & dosificación , Vacuna BNT162/efectos adversos , Femenino , Humanos , Inmunidad Celular , Inmunidad Humoral , Inmunoglobulina G/biosíntesis , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , SARS-CoV-2/inmunología , Glicoproteína de la Espiga del Coronavirus/inmunología , Linfocitos T/inmunología , Adulto JovenRESUMEN
Primary immunodeficiency (PID) diseases result from genetic defects of the immune system that increase a patient's susceptibility to infections. The types of infections that occur in patients with PID diseases are dictated largely by the nature of the immunodeficiency, which can be defined by dysfunction of cellular or humoral defenses. An increasing number of PID diseases, including those with both cellular and humoral defects, have antibody deficiency as a major feature, and as a result can benefit from immunoglobulin replacement therapy. In fact, the most common PID diseases worldwide are antibody deficiencies and include common variable immunodeficiency, congenital agammaglobulinemia, hyper-IgM syndrome, specific antibody deficiency, and Good syndrome. Although immunoglobulin replacement therapy is the cornerstone of treatment for the majority of these conditions, a thorough understanding of the specific infections for which these patients are at increased risk can hasten diagnosis and guide additional therapies. Moreover, the infection trends in some patients with PID disease who have profound defects of cellular immunity, such as autosomal-dominant hyper-IgE syndrome (Job/Buckley syndrome) or dedicator of cytokinesis 8 (DOCK8) deficiency, suggest that select patients might benefit from immunoglobulin replacement therapy even if their immunodeficiency is not limited to antibody defects. In this review, we provide an overview of the predisposition to infections seen in PID disease that may benefit from immunoglobulin replacement therapy.
Asunto(s)
Inmunoglobulinas/uso terapéutico , Síndromes de Inmunodeficiencia/complicaciones , Síndromes de Inmunodeficiencia/terapia , Infecciones/inmunología , Agammaglobulinemia/complicaciones , Agammaglobulinemia/inmunología , Agammaglobulinemia/terapia , Inmunodeficiencia Variable Común/inmunología , Inmunodeficiencia Variable Común/terapia , Enfermedades Genéticas Ligadas al Cromosoma X/inmunología , Enfermedades Genéticas Ligadas al Cromosoma X/terapia , Humanos , Síndromes de Inmunodeficiencia/clasificación , Síndromes de Inmunodeficiencia/inmunología , Infecciones/terapia , Factores de RiesgoRESUMEN
BACKGROUND: Colonization by Aspergillus fumigatus in patients with cystic fibrosis (CF) can cause A fumigatus sensitization and/or allergic bronchopulmonary aspergillosis (ABPA), which affects pulmonary function and clinical outcomes. Recent studies show that specific allergens upregulate the surface-expressed basophil marker CD203c in sensitized subjects, a response that can be readily measured by using flow cytometry. OBJECTIVE: We sought to identify A fumigatus sensitization in patients with CF by using the basophil activation test (BAT). METHODS: Patients with CF attending Beaumont Hospital were screened for study inclusion. BAT was used to identify A fumigatus sensitization. Serologic (total and A fumigatus-specific IgE), pulmonary function, and body mass index measurements were performed. RESULTS: The BAT discriminates A fumigatus-sensitized from nonsensitized patients with CF. Persistent isolation of A fumigatus in sputum is a significant risk factor for A fumigatus sensitization. Levels of the A fumigatus-stimulated basophil activation marker CD203c inversely correlated with pulmonary function and body mass index in A fumigatus-sensitized but not nonsensitized patients with CF. Total and A fumigatus-specific IgE, but not IgG, levels are increased in A fumigatus-sensitized patients with CF and ABPA when compared with those in A fumigatus-sensitized and nonsensitized patients with CF without ABPA. Itraconazole treatment did not affect A fumigatus sensitization. CONCLUSION: Combining the BAT with routine serologic testing allows classification of patients with CF into 3 groups: nonsensitized, A fumigatus-sensitized, and ABPA. Accurate and prompt identification of A fumigatus-associated clinical status might allow early and targeted therapeutic intervention, potentially improving clinical outcomes.
Asunto(s)
Aspergilosis/etiología , Aspergilosis/metabolismo , Aspergillus/inmunología , Basófilos/metabolismo , Fibrosis Quística/complicaciones , Hidrolasas Diéster Fosfóricas/metabolismo , Pirofosfatasas/metabolismo , Especificidad de Anticuerpos/inmunología , Antifúngicos/farmacología , Antifúngicos/uso terapéutico , Antígenos Fúngicos/inmunología , Antígenos de Superficie/metabolismo , Aspergilosis/diagnóstico , Aspergilosis/tratamiento farmacológico , Basófilos/inmunología , Biomarcadores , Índice de Masa Corporal , Fibrosis Quística/fisiopatología , Femenino , Volumen Espiratorio Forzado , Humanos , Inmunoglobulina E/sangre , Inmunoglobulina E/inmunología , Inmunoglobulina G/sangre , Inmunoglobulina G/inmunología , Inmunofenotipificación , Recuento de Leucocitos , Masculino , Estudios Prospectivos , Esputo/inmunología , Esputo/microbiologíaRESUMEN
The diagnosis of cystic fibrosis (CF) patients with allergic bronchopulmonary aspergillosis (ABPA) is clinically challenging, due to the absence of an objective biological test. Since blood basophils play a major role in allergic responses, we hypothesised that changes in their surface activation pattern discriminate between CF patients with and without ABPA.We conducted a prospective longitudinal study (Stanford cohort) comparing basophil activation test CD203c levels by flow cytometry before and after activation with Aspergillus fumigatus allergen extract or recombinant Asp f1 in 20 CF patients with ABPA (CF-ABPA) and in two comparison groups: CF patients with A. fumigatus colonisation (AC) but without ABPA (CF-AC; n=13) and CF patients without either AC or ABPA (CF; n=12). Patients were tested every 6â months and when ill with pulmonary exacerbation. We also conducted cross-sectional validation in a separate patient set (Dublin cohort).Basophil CD203c surface expression reliably discriminated CF-ABPA from CF-AC and CF over time. Ex vivo stimulation with A. fumigatus extract or recombinant Asp f1 produced similar results within the Stanford (p<0.0001) and the Dublin cohorts. CF-ABPA patients were likelier to have elevated specific IgE to A. fumigatus and were less frequently co-infected with Staphylococcus aureus.Basophil CD203c upregulation is a suitable diagnostic and stable monitoring biomarker of ABPA in CF.
Asunto(s)
Aspergilosis Broncopulmonar Alérgica/metabolismo , Basófilos/metabolismo , Portador Sano/metabolismo , Fibrosis Quística/metabolismo , Hidrolasas Diéster Fosfóricas/metabolismo , Aspergilosis Pulmonar/metabolismo , Pirofosfatasas/metabolismo , Adolescente , Adulto , Alérgenos/farmacología , Aspergilosis Broncopulmonar Alérgica/complicaciones , Aspergilosis Broncopulmonar Alérgica/diagnóstico , Aspergillus fumigatus/inmunología , Basófilos/efectos de los fármacos , Biomarcadores/metabolismo , Portador Sano/diagnóstico , Estudios de Casos y Controles , Niño , Estudios de Cohortes , Fibrosis Quística/complicaciones , Fibrosis Quística/diagnóstico , Femenino , Citometría de Flujo , Humanos , Masculino , Estudios Prospectivos , Aspergilosis Pulmonar/complicaciones , Aspergilosis Pulmonar/diagnóstico , Adulto JovenAsunto(s)
Antígeno CTLA-4/genética , Síndromes de Inmunodeficiencia/diagnóstico , Pneumocystis carinii/fisiología , Neumonía por Pneumocystis/genética , Adolescente , Haploinsuficiencia , Humanos , Huésped Inmunocomprometido , Síndromes de Inmunodeficiencia/genética , Masculino , Osteomielitis , Neumonía por Pneumocystis/diagnósticoAsunto(s)
Anticuerpos Monoclonales Humanizados/uso terapéutico , Micosis/tratamiento farmacológico , Rinitis Alérgica Perenne/tratamiento farmacológico , Sinusitis/tratamiento farmacológico , Adolescente , Hongos/aislamiento & purificación , Humanos , Imagen por Resonancia Magnética , Masculino , Micosis/diagnóstico , Rinitis Alérgica Perenne/diagnóstico , Rinitis Alérgica Perenne/microbiología , Sinusitis/diagnóstico , Sinusitis/microbiología , Tomografía Computarizada por Rayos XAsunto(s)
Antirreumáticos/uso terapéutico , Enfermedades Autoinmunes/tratamiento farmacológico , Enfermedades Autoinmunes/genética , Proteína Antagonista del Receptor de Interleucina 1/uso terapéutico , Síndrome de Activación Macrofágica/genética , Proteína de Unión al GTP cdc42/genética , Niño , Femenino , Humanos , Lactante , Inflamación/genética , Interleucina-1beta/antagonistas & inhibidores , MasculinoRESUMEN
WHIM (Warts, Hypogammaglobulinemia, Infections, Myelokathexis) syndrome is a rare primary immunodeficiency predominantly caused by heterozygous gain-of-function mutations in the c-terminus of the gene CXCR4. These CXCR4 variants display impaired receptor trafficking with persistence of the CXCR4 receptor on the surface resulting in hyperactive downstream signaling following CXCL12 stimulation. In turn, this results in defective lymphoid differentiation, and reduced blood neutrophil and lymphocyte numbers. Here we report a CXCR4 mutation that in two members of a kindred, led to life-long autoimmunity and lymphoid hypertrophy as the primary clinical manifestations of WHIM syndrome. We examine the functional effects of this mutation, and how these have affected phosphorylation, activation, and receptor internalization.
RESUMEN
Biallelic mutations in the ACP5 gene cause spondyloenchondrodysplasia with immune dysregulation (SPENCDI). SPENCDI is characterized by the phenotypic triad of skeletal dysplasia, innate and adaptive immune dysfunction, and variable neurologic findings ranging from asymptomatic brain calcifications to severe developmental delay with spasticity. Immune dysregulation in SPENCDI is often refractory to standard immunosuppressive treatments. Here, we present the cases of two patients with SPENCDI and recalcitrant autoimmune cytopenias who demonstrated a favorable clinical response to targeted JAK inhibition over a period of more than 3 years. One of the patients exhibited steadily rising IgG levels and a bone marrow biopsy revealed smoldering multiple myeloma. A review of the literature uncovered that approximately half of the SPENCDI patients reported to date exhibited increased IgG levels. Screening for multiple myeloma in SPENCDI patients with rising IgG levels should therefore be considered.
Asunto(s)
Anemia Hemolítica Autoinmune , Enfermedades Autoinmunes , Inmunoglobulina G , Síndromes de Inmunodeficiencia , Janus Quinasa 2 , Osteocondrodisplasias , Trombocitopenia , Humanos , Fosfatasa Ácida Tartratorresistente/genética , Janus Quinasa 1RESUMEN
Studies of the monogenic autoimmune disease immunodysregulation polyendocrinopathy enteropathy X-linked syndrome (IPEX) have elucidated the essential function of the transcription factor FOXP3 and thymic-derived regulatory T cells (Tregs) in controlling peripheral tolerance. However, the presence and the source of autoreactive T cells in IPEX remain undetermined. Here, we investigated how FOXP3 deficiency affects the T cell receptor (TCR) repertoire and Treg stability in vivo and compared T cell abnormalities in patients with IPEX with those in patients with autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy syndrome (APECED). To study Tregs independently of their phenotype and to analyze T cell autoreactivity, we combined Treg-specific demethylation region analyses, single-cell multiomic profiling, and bulk TCR sequencing. We found that patients with IPEX, unlike patients with APECED, have expanded autoreactive T cells originating from both autoreactive effector T cells (Teffs) and Tregs. In addition, a fraction of the expanded Tregs from patients with IPEX lost their phenotypic and functional markers, including CD25 and FOXP3. Functional experiments with CRISPR-Cas9-mediated FOXP3 knockout Tregs and Tregs from patients with IPEX indicated that the patients' Tregs gain a TH2-skewed Teff-like function, which is consistent with immune dysregulation observed in these patients. Analyses of FOXP3 mutation-carrier mothers and a patient with IPEX after hematopoietic stem cell transplantation indicated that Tregs expressing nonmutated FOXP3 prevent the accumulation of autoreactive Teffs and unstable Tregs. These findings could be directly used for diagnostic and prognostic purposes and for monitoring the effects of immunomodulatory treatments.
Asunto(s)
Enfermedades Genéticas Ligadas al Cromosoma X , Poliendocrinopatías Autoinmunes , Humanos , Poliendocrinopatías Autoinmunes/genética , Poliendocrinopatías Autoinmunes/terapia , Enfermedades Genéticas Ligadas al Cromosoma X/genética , Linfocitos T Reguladores , Mutación/genética , Síndrome , Factores de Transcripción Forkhead/genética , Receptores de Antígenos de Linfocitos T/genéticaRESUMEN
The mammalian target of rapamycin (mTOR) pathway contributes to various immunoinflammatory processes. Yet, its potential involvement in basophil responses in allergy remains unclear. In this pilot study, we quantified two key mTOR effector phosphoproteins, the eukaryotic initiation factor 4E (peIF4E) and S6 ribosomal protein (pS6rp), in blood basophils from nut allergy patients (NA, N = 16) and healthy controls (HC, N = 13). Without stimulation in vitro, basophil peIF4E levels were higher in NA than HC subjects (P = 0.014). Stimulation with nut (offending) but not chicken / rice (non-offending) extract increased basophil peIF4E and pS6rp levels (+32%, P = 0.018, and +98%, P = 0.0026, respectively) in NA but not HC subjects, concomitant with increased surface levels of CD203c and CD63, both known to reflect basophil activation. Pre-treatment with the mTOR inhibitor rapamycin decreased pS6rp and CD203c responses in nut extract-stimulated basophils in NA subjects. Thus, basophil responses to offending allergens are associated with modulation of mTOR effector phosphoproteins.
Asunto(s)
Basófilos/inmunología , Factor 4E Eucariótico de Iniciación/inmunología , Hipersensibilidad a la Nuez/inmunología , Hipersensibilidad al Cacahuete/inmunología , Proteína S6 Ribosómica/inmunología , Serina-Treonina Quinasas TOR/inmunología , Adolescente , Alérgenos/inmunología , Alérgenos/farmacología , Anacardium/inmunología , Basófilos/efectos de los fármacos , Niño , Preescolar , Femenino , Humanos , Juglans/inmunología , Masculino , Hipersensibilidad a la Nuez/sangre , Hipersensibilidad al Cacahuete/sangre , Fosforilación , Extractos Vegetales/inmunología , Extractos Vegetales/farmacología , Sirolimus/farmacología , Serina-Treonina Quinasas TOR/antagonistas & inhibidores , Adulto JovenAsunto(s)
Angioedema/tratamiento farmacológico , Angioedemas Hereditarios/tratamiento farmacológico , Proteínas Inactivadoras del Complemento 1/deficiencia , Linfoma/tratamiento farmacológico , Paraproteinemias/tratamiento farmacológico , Rituximab/uso terapéutico , Anciano de 80 o más Años , Angioedema/genética , Angioedema/inmunología , Angioedema/patología , Angioedemas Hereditarios/genética , Angioedemas Hereditarios/inmunología , Angioedemas Hereditarios/patología , Antiinflamatorios no Esteroideos/uso terapéutico , Bradiquinina/análogos & derivados , Bradiquinina/uso terapéutico , Antagonistas del Receptor de Bradiquinina B2/uso terapéutico , Proteínas Inactivadoras del Complemento 1/genética , Proteínas Inactivadoras del Complemento 1/inmunología , Femenino , Humanos , Factores Inmunológicos/uso terapéutico , Linfoma/genética , Linfoma/inmunología , Linfoma/patología , Masculino , Persona de Mediana Edad , Paraproteinemias/genética , Paraproteinemias/inmunología , Paraproteinemias/patología , Péptidos/uso terapéutico , Resultado del TratamientoRESUMEN
Cystic fibrosis (CF) patients undergo progressive airway destruction caused in part by chronic neutrophilic inflammation. While opportunistic pathogens infecting CF airways can cause inflammation, we hypothesized that host-derived metabolic and stress signals would also play a role in this process. We show that neutrophils that have entered CF airways have increased phosphorylation of the eukaryotic initiation factor 4E and its partner the 4E-binding protein 1; 2 key effectors in the growth factor- and amino acid-regulated mammalian target of rapamycin (mTOR) pathway. Furthermore CF airway neutrophils display increased phosphorylation of the cAMP response element binding protein (CREB), a major transcriptional coactivator in stress signaling cascades. These active intracellular pathways are associated with increased surface expression of critical adaptor molecules, including the growth factor receptor CD114 and the receptor for advanced glycation end-products (RAGE), a CREB inducer and sensor for host-derived damage-associated molecular patterns (DAMPs). Most CF airway fluids lack any detectable soluble RAGE, an inhibitory decoy receptor for DAMPs. Concomitantly, CF airway fluids displayed high and consequently unopposed levels of S100A12; a potent mucosa- and neutrophil-derived DAMP. CF airway neutrophils also show increased surface levels of 2 critical CREB targets, the purine-recycling enzyme CD39 and the multifunctional, mTOR-inducing CXCR4 receptor. This coordinated set of events occurs in all patients, even in the context of minimal airway inflammation and well-preserved lung function. Taken together, our data demonstrate an early and sustained activation of host-responsive metabolic and stress pathways upon neutrophil entry into CF airways, suggesting potential targets for therapeutic modulation.
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Quimiotaxis de Leucocito , Fibrosis Quística/inmunología , Fibrosis Quística/metabolismo , Neutrófilos/inmunología , Proteína de Unión a Elemento de Respuesta al AMP Cíclico , Productos Finales de Glicación Avanzada , Humanos , Enfermedades Pulmonares/inmunología , Enfermedades Pulmonares/metabolismo , Estrés Oxidativo , Fosforilación , Proteínas Quinasas , Transducción de Señal , Serina-Treonina Quinasas TORRESUMEN
Primary immunodeficiency may present with treatment-refractory enteropathy. We present two patients with celiac/celiac-like disease diagnosed in early childhood and refractory to the gluten-free diet. One patient had features of multi-system autoimmunity, whereas the other had celiac-like disease as an isolated clinical finding. Both patients underwent genetic testing given disease refractoriness and were ultimately diagnosed with cytotoxic T lymphocyte antigen 4 (CTLA4) haploinsufficiency. They are both now in complete clinical and endoscopic remission on abatacept. CTLA4 haploinsufficiency has incomplete penetrance and significant phenotypic heterogeneity but should be considered in the differential diagnosis of refractory celiac/celiac-like disease, as treatment implications are significant.
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Enfermedad Celíaca , Autoinmunidad , Antígeno CTLA-4/genética , Enfermedad Celíaca/diagnóstico , Enfermedad Celíaca/genética , Preescolar , Dieta Sin Gluten , Haploinsuficiencia , HumanosRESUMEN
RATIONALE: Basophils contribute to anaphylaxis and allergies. We examined the utility of assessing basophil-associated surface antigens (CD11b/CD63/CD123/CD203c/CD294) in characterizing and monitoring subjects with nut allergy. METHODS: We used flow cytometry to analyze basophils at baseline (without any activation) and after ex vivo stimulation of whole blood by addition of nut or other allergens for 2, 10, and 30 min. We also evaluated whether basophil expression of CD11b/CD63/CD123/CD203c/CD294 was altered in subjects treated with anti-IgE monoclonal antibody (omalizumab) to reduce plasma levels of IgE. RESULTS: We demonstrate that basophil CD203c levels are increased at baseline in subjects with nut allergy compared to healthy controls (13 subjects in each group, p < 0.0001). Furthermore, we confirm that significantly increased expression of CD203c occurs on subject basophils when stimulated with the allergen to which the subject is sensitive and can be detected rapidly (10 min of stimulation, n = 11, p < 0.0008). In 5 subjects with severe peanut allergy, basophil CD203c expression following stimulation with peanut allergen was significantly decreased (p < 0.05) after 4 and 8 weeks of omalizumab treatment but returned toward pretreatment levels after treatment cessation. CONCLUSIONS: Subjects with nut allergy show an increase of basophil CD203c levels at baseline and following rapid ex vivo stimulation with nut allergen. Both can be reduced by omalizumab therapy. These results highlight the potential of using basophil CD203c levels for baseline diagnosis and therapeutic monitoring in subjects with nut allergy.
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Antialérgicos/uso terapéutico , Anticuerpos Monoclonales/uso terapéutico , Basófilos/metabolismo , Hipersensibilidad a la Nuez/metabolismo , Hidrolasas Diéster Fosfóricas/biosíntesis , Pirofosfatasas/biosíntesis , Adolescente , Adulto , Anticuerpos Antiidiotipos , Anticuerpos Monoclonales Humanizados , Basófilos/efectos de los fármacos , Basófilos/inmunología , Biomarcadores/análisis , Biomarcadores/sangre , Separación Celular , Niño , Preescolar , Femenino , Citometría de Flujo , Humanos , Lactante , Masculino , Hipersensibilidad a la Nuez/tratamiento farmacológico , Hipersensibilidad a la Nuez/inmunología , Omalizumab , Hidrolasas Diéster Fosfóricas/efectos de los fármacos , Pirofosfatasas/efectos de los fármacos , Sensibilidad y Especificidad , Adulto JovenRESUMEN
BACKGROUND AND AIM: Eosinophilic esophagitis (EoE) is a chronic inflammatory disorder characterized by upper gastrointestinal symptoms and the presence of high numbers of eosinophils in the esophagus. Although eosinophils in the esophagus have been found to be activated in subjects with EoE, detailed studies of intracellular signaling pathways involved in the mechanism of activation of eosinophils in EoE have heretofore been limited. The aim of the study was to assess whether any surface molecules or transcription factors are activated in peripheral eosinophils in subjects with EoE. METHODS: Eosinophils and CD3+ lymphocytes were identified directly from 50 µL of whole blood of EoE and control subjects. Using Hi-FACS, levels of surface activation markers, including CD66b, and intracellular phosphoepitopes, including phosphorylated forms of signal transducer and activator of transcription (phospho-STAT) 1 and 6, were measured within each cell subset. RESULTS: Levels of surface CD66b as well as levels of intracellular phospho-STAT1 and phospho-STAT6 in peripheral blood eosinophils were significantly higher for untreated subjects with EoE vs healthy controls (P < 0.05). Levels of phospho-STAT1 and phospho-STAT6 in peripheral blood eosinophils were lower in subjects with EoE on therapy versus untreated subjects with EoE (P < 0.05). CONCLUSIONS: Levels of phospho-STAT1 and phospho-STAT6, transcription factors involved in inflammatory processes, were both significantly higher in peripheral eosinophils from untreated (ie, newly diagnosed) subjects with EoE versus subjects with EoE on therapy, healthy controls. Blood-based measurements of CD66b and phospho-STAT levels in peripheral eosinophils may be beneficial for identifying EoE.
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Esofagitis Eosinofílica/inmunología , Eosinófilos/inmunología , Activación de Linfocitos , Adolescente , Antígenos CD/metabolismo , Complejo CD3/metabolismo , Moléculas de Adhesión Celular/metabolismo , Niño , Preescolar , Esofagitis Eosinofílica/metabolismo , Esofagitis Eosinofílica/terapia , Eosinófilos/metabolismo , Femenino , Proteínas Ligadas a GPI/metabolismo , Humanos , Inmunofenotipificación , Terapia de Inmunosupresión , Linfocitos/metabolismo , Masculino , Fosforilación , Procesamiento Proteico-Postraduccional , Factor de Transcripción STAT1/genética , Factor de Transcripción STAT1/metabolismo , Factor de Transcripción STAT6/genética , Factor de Transcripción STAT6/metabolismo , Índice de Severidad de la Enfermedad , Adulto JovenRESUMEN
Blood neutrophils recruited to cystic fibrosis (CF) airways are believed to be rapidly killed by resident bacteria and to passively release elastase and other toxic by-products that promote disease progression. By single-cell analysis, we demonstrate that profound functional and signaling changes readily occur within viable neutrophils recruited to CF airways, compared with their blood counterparts. Airway neutrophils have undergone conventional activation, as shown by decreased intracellular glutathione, increased lipid raft assembly, surface mobilization of CD11b+ and CD66b+ granules, and increased levels of the cytoskeleton-associated phospho-Syk kinase. Unexpectedly, they also mobilize to the surface CD63+ elastase-rich granules, usually confined intracellularly, and lose surface expression of CD16 and CD14, both key receptors in phagocytosis. Furthermore, they express CD80, major histocompatibility complex type II, and the prostaglandin D2 receptor CD294, all normally associated with other lineages, which reflects functional reprogramming. This notion is reinforced by their decreased total phosphotyrosine levels, mirroring a postactivated stage, and increased levels of the phospho-S6 ribosomal protein, a key anabolic switch. Thus, we identified a subset of neutrophils within CF airways with a viable but dysfunctional phenotype. This subset provides a possible therapeutic target and indicates a need to revisit current paradigms of CF airway disease.