Functional Immunophenotyping of Children with Critical Status Asthmaticus Identifies Differential Gene Expression Responses in Neutrophils Exposed to a Poly(I:C) Stimulus
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PROVIDER: PRJNA844008 | ENA |
REPOSITORIES: ENA
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Project description:We determined whether we could identify clusters of children with critical asthma by functional immunophenotyping using an intracellular viral analog stimulus. We performed a single-center, prospective, observational cohort study of 43 children ages 6 – 17 years admitted to a pediatric intensive care unit for an asthma attack between July 2019 to February 2021.
2022-06-05 | GSE205151 | GEO
Project description:Preschool children with recurrent wheezing are heterogeneous, with differing responses to respiratory viral infections. Although neutrophils are crucial for host defense, their function has not been studied in this population. We performed functional immunophenotyping on isolated blood neutrophils from 52 preschool children with recurrent wheezing (aeroallergen sensitization, n=16; no sensitization, n=36). Blood neutrophils were purified and cultured overnight with polyinosinic:polycytidylic acid (poly(I:C)) as a viral analog stimulus. Neutrophils underwent next-generation sequencing with Reactome pathway analysis and were analyzed for cytokine secretion, apoptosis, myeloperoxidase and extracellular DNA release. CD14+ monocytes were also exposed to neutrophil culture supernatant and analyzed for markers of M1 and M2 activation. 495 genes, related largely to the innate immune system and neutrophil degranulation, were differently expressed in children with versus without aeroallergen sensitization. Functional experiments identified more neutrophil degranulation and extracellular trap formation (i.e., more myeloperoxidase and extracellular DNA) and less neutrophil pro-inflammatory cytokine secretion in children with aeroallergen sensitization. Neutrophils also shifted CD14+ monocytes to a more anti-inflammatory (i.e., M2) phenotype in sensitized children and a more pro-inflammatory (i.e., M1) phenotype in non-sensitized children. Although both groups experienced viral exacerbations, annualized exacerbation rates prompting unscheduled healthcare were also higher in children without aeroallergen sensitization after enrollment. Systemic neutrophil responses to viral infection differ by allergic phenotype and may be less effective in preschool children without allergic inflammation. Further studies of neutrophil function are needed in this population, which often has less favorable therapeutic responses to inhaled corticosteroids and other therapies directed at T2-high inflammation.
2024-01-29 | GSE254019 | GEO
Project description:The host immune response to a viral immune stimulus has not been examined in children during a life-threatening asthma attack. We determined whether we could identify clusters of children with critical asthma by functional immunophenotyping using an intracellular viral analog stimulus. We performed a single-center, prospective, observational cohort study of 43 children ages 6-17 years admitted to a pediatric intensive care unit for an asthma attack between July 2019 to February 2021. Neutrophils were isolated from children, stimulated overnight with LyoVec poly(I:C), and mRNA was analyzed using a targeted Nanostring immunology array. Network analysis of the differentially expressed transcripts for the paired LyoVec poly(I:C) samples was performed. We identified two clusters by functional immunophenotyping that differed by the Asthma Control Test score. Cluster 1 (n = 23) had a higher proportion of children with uncontrolled asthma in the four weeks prior to PICU admission compared with cluster 2 (n = 20). Pathways up-regulated in cluster 1 versus cluster 2 included chemokine receptor/chemokines, interleukin-10 (IL-10), IL-4, and IL-13 signaling. Larger validation studies and clinical phenotyping of children with critical asthma are needed to determine the predictive utility of these clusters in a larger clinical setting.
| S-EPMC9666940 | biostudies-literature
Project description:Functional immunophenotyping of blood neutrophils identifies novel endotypes of viral response in preschool children with recurrent wheezing
| PRJNA1068130 | ENA
Project description:Utilizing a chronic colitis model in Apoe-deficient mice, we identify the crucial role of neutrophils in the development of atherosclerosis. We demonstrate that neutrophils are the first cells to respond in both the circulation and the aorta to the repetitive stimulus of chronic intestinal inflammation and acquire inflammatory, pathogenic phenotype. Single cell RNA-Seq and single cell ATAC-Seq analysis of sorted GMP popu;ation revealed that circulating bone marrow neutrophils from repeated DSS-exposed mice acquire the pathogenic phenotype in the bone marrow which can accelerate vascular inflammation in turn.
2025-03-25 | GSE261993 | GEO
Project description:Utilizing a chronic colitis model in Apoe-deficient mice, we identify the crucial role of neutrophils in the development of atherosclerosis. We demonstrate that neutrophils are the first cells to respond in both the circulation and the aorta to the repetitive stimulus of chronic intestinal inflammation and acquire inflammatory, pathogenic phenotype. Single cell RNA-Seq and single cell ATAC-Seq analysis of sorted GMP popu;ation revealed that circulating bone marrow neutrophils from repeated DSS-exposed mice acquire the pathogenic phenotype in the bone marrow which can accelerate vascular inflammation in turn.
2025-03-25 | GSE261992 | GEO
Project description:Polymorphonuclear cells (neutrophils) play an important role in the systemic inflammatory response syndrome and the development of sepsis. These cells are essential for the defense against microorganisms, but may also cause tissue damage. Therefore, neutrophil numbers and activity are considered to be tightly regulated. Previous studies have investigated gene transcription during experimental endotoxemia in whole blood and peripheral blood mononuclear cells. However, the gene transcription response of the circulating pool of neutrophils to systemic inflammatory stimulation in vivo is currently unclear. We examined neutrophil gene transcription kinetics in healthy human subjects (n=4) administered a single dose of endotoxin (LPS, 2 ng/kg iv). In addition, freshly isolated neutrophils were stimulated ex vivo with LPS, TNFM-NM-1, G-CSF and GM-CSF to identify stimulus-specific gene transcription responses. Whole transcriptome microarray analysis of circulating neutrophils at 2, 4 and 6 hours after LPS infusion revealed activation of inflammatory networks which are involved in signaling of TNFM-NM-1 and IL-1M-NM-1 and IL-1M-NM-2. The transcriptome profile of inflammatory activated neutrophils in vivo reflects extended survival and regulation of inflammatory responses. We show that these changes in neutrophil transcriptome are most likely due to a combination of early activation of circulating neutrophils by TNFM-NM-1 and G-CSF and a mobilization of young neutrophils from the bone marrow. After LPS infusion blood was taken at t=0, t=2, t=4 and t=6 hours. Neutrophils were isolated and gene expression of these cells was assessed. T=2, t=4 and t=6 were related to t=0 as control condition
2012-06-04 | E-GEOD-35590 | biostudies-arrayexpress
Project description:The innate mechanisms associated with viral exacerbations in preschool children with recurrent wheezing are not understood. We assessed differential gene expression in blood neutrophils from preschool children with recurrent wheezing, stratified by aeroallergen sensitization, at baseline and after exposure to polyinosinic-polycytidylic acid (poly(I:C)). We also examined whether poly(I:C)-stimulated blood neutrophils influenced airway epithelial gene expression. Blood neutrophils were purified and cultured overnight with poly(I:C) and underwent next-generation sequencing with Reactome pathway analysis. Primary human small airway epithelial cells were treated with poly(I:C)-treated neutrophil culture supernatants and were analyzed for Type 1 interferon gene expression with a targeted array. Symptoms and exacerbations were assessed in participants over 12 months. 436 genes were differently expressed in neutrophils from children with versus without aeroallergen sensitization at baseline, with significant downregulation of Type 1 interferons. These Type 1 interferons were significantly upregulated in sensitized children after poly(I:C) stimulation. Confirmatory experiments demonstrated similar upregulation of Type 1 interferons in IL-4 treated neutrophils stimulated with poly(I:C). Poly(I:C)-treated neutrophil supernatants from children with aeroallergen sensitization also induced a Type 1 interferon response in epithelial cells. Children with aeroallergen sensitization also had higher symptom scores during exacerbations and these symptom differences persisted for three days after prednisolone treatment. We conclude that Type 1 interferon responses are dysregulated in preschool children with aeroallergen sensitization, which is in turn associated with exacerbation severity. Given the importance of Type 1 interferon signaling in viral resolution, additional studies of neutrophil Type 1 interferon responses are needed in this population.
2025-03-10 | GSE261070 | GEO
Project description:One of the key impacts of probiotic bacteria is the ability to beneficially modulate innate and adaptive immune responses of various cell types of the gut-associated lymphoid tissue. In this study we used an in vitro intestinal epithelial cell model to investigate the impact of three probiotic bacteria Lactobacillus helveticus R0052, Bifidobacterium longum subsp. infantis R0033 and Bifidobacterium bifidum R0071 individually, in combination (R0033, R0052 and R0071) and of a specific surface-layer protein partially purified from L. helveticus R0052 (R0052-SLP) in response to a known dsRNA pro-inflammatory stimulus, poly (I:C). Genome-wide human expression microarray chips were used to evaluate other cellular pathways and gain a deeper insight into global gene and pathway modulation. Here we report that the challenge by poly (I:C)-only resulted in the differential expression of 863 genes; whereas, the co-challenge with either R0033, R0052, R0071, R0052-SLP and the multi-strain combination (R0052, R0033 and R0071) resulted in differential expression of 467 genes, 369 genes, 293 genes, 219 genes and 132 genes; respectively. The multi-strain combination had a greater impact than each single component strain at reducing the number of genes modulated. Among the major human pathways modulated by probiotics or R0052-SLP in response to poly (I:C) include: immune/virus, cellular/signaling, nervous/endocrine and autoimmune disorder related pathways. Cytokine and chemokine profiling was performed at the protein level based on specific markers from the TNF-α and NF-KB signaling pathways. The results revealed single strain, multi-strain and R0052-SLP specific attenuation for the majority of proteins measured (TNF-α, IL-8, CXCL1, CXCL2, CXCL10 and IL-10) indicating potentially different mechanisms for each strain, multi-strain and protein component and reinforcing the rationale for multi-strain combinations. The overall study design consisted of 8 different treatments/challenges of HT-29. Briefly, HT-29 cells were co-challenge for 3 h with either the single strains (R0033, R0052 or R0071) or probiotic combination (PC) alone or in combination with poly (I:C) at 10 µg/mL. Also, a specific surface layer protein purified from R0052 (R0052-SLP) was also co-challenged with poly (I:C) in HT-29 cells. Treatment/challenges of poly (I:C)-only, surface layer protein-only (R0052-SLP-only) and probiotic combination-only or PC-only were also evaluated. All challenges and co-challenges were compared to unchallenged control HT-29 cells. 4 biological reps were performed for each of the 8 samples/treatments.
2015-08-25 | E-GEOD-71515 | biostudies-arrayexpress
Project description:Neutrophils play a key role in the innate immunity and the first line of defense against invading pathogens. The aim of this work is to investigate the neutrophil responses and transcriptomic alteration in the porcine peripheral blood by employing Poly I:C to emulate viral infection. The pigs with the two-tailed critical value of neutrophilia post 4h with poly I:C stimulation from a Duroc-Erhualian F2 population.
2017-08-30 | E-MTAB-4928 | biostudies-arrayexpress