Assessing Phenotypic Heterogeneity in Intestinal Tissue Eosinophils

Olbrich, Courtney L; Larsen, Leigha D; Spencer, Lisa A.

doi:10.1007/978-1-0716-1095-4_19

Cited by 4 publications

(4 citation statements)

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“…Single‐cell LP and IE suspensions were obtained from intestinal tissues as previously described 26 . Briefly, whole small intestine was collected, Peyer's patches excised, and tissue pieces incubated in mucolytic DTT‐containing buffer (1 mM dl ‐dithiothreitol, 10% FBS, 1 mM HEPES, 2.5 mM NaHCO3 in HBSS −/− ) to release IE cells.…”

Section: Methodsmentioning

confidence: 99%

“…LP cells were recovered by digesting tissue pieces in a collagenase containing buffer (175 U/ml collagenase type 1, 10% FBS, 0.5 mM CaCl 2 , 0.5 mM MgCl 2 , 1× Pen‐Strep in RPMI 1640 with l ‐glutamine and 25 mM HEPES) prior to mechanical disruption and passage through a 70 μM cell strainer. Both IE and LP cell preparations were further purified by discontinuous Percoll gradient centrifugation as described 26 . Briefly, cell pellets were resuspended in Percoll (Cytiva, Upsala, Sweden) diluted to 44% in RPMI containing phenol red and underlaid with Percoll diluted to 67% in RPMI without phenol red prior to centrifugation at 1000 × g .…”

Section: Methodsmentioning

confidence: 99%

“…Single-cell LP and IE suspensions were obtained from intestinal tissues as previously described. 26 Briefly, whole small intestine was collected, Peyer's patches excised, and tissue pieces incubated in mucolytic DTTcontaining buffer (1 mM DL-dithiothreitol, 10% FBS, 1 mM HEPES, 26 Briefly, cell pellets were resuspended in Percoll (Cytiva, Upsala, Sweden) diluted to 44% in RPMI containing phenol red and underlaid with Percoll diluted to 67% in RPMI without phenol red prior to centrifugation at 1000 × g. Leukocytes were collected from the interface between 44% and 67% layers.…”

Section: Recovery Of Intestinal Leukocytes From Lp and Ie Compartmentsmentioning

confidence: 99%

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Modulation of surface CD11c expression tracks plasticity in murine intestinal tissue eosinophils

Larsen

Dockstader

Olbrich

et al. 2022

Journal of Leukocyte Biology

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Intestinal eosinophils are implicated in the inflammatory pathology of eosinophilic gastrointestinal diseases and inflammatory bowel diseases. Eosinophils also contribute to intestinal immunologic and tissue homeostasis and host defense. Recent studies in allergic airway disease suggest functional subphenotypes of eosinophils may underly their pathogenic versus protective roles. However, subphenotypes of intestinal eosinophils have not been defined and are complicated by their constitutive expression of the putative eosinophil inflammatory marker CD11c. Here, we propose a framework for subphenotype characterization of intestinal eosinophils based on relative intensity of surface CD11c expression. Using this flow cytometry framework in parallel with histology and BrdU tracing, we characterize intestinal eosinophil subphenotypes and monitor their plasticity at baseline and within the context of acute allergic and chronic systemic inflammation. Data reveal a conserved continuum of CD11c expression amongst intestinal eosinophils in health and acute disease states that overall tracked with other markers of activation. Oral allergen challenge induced recruitment of eosinophils into small intestinal lamina propria surrounding crypts, followed by in situ induction of CD11c expression in parallel with eosinophil redistribution into intestinal villi. Allergen challenge also elicited eosinophil transepithelial migration and the appearance of CD11cloCD11bhi eosinophils in the intestinal lumen. Chronic inflammation driven by overexpression of TNFα led to a qualitative shift in the relative abundance of CD11c‐defined eosinophil subphenotypes favoring CD11chi‐expressing eosinophils. These findings provide new insights into heterogeneity of intestinal tissue eosinophils and offer a framework for measuring and tracking eosinophil subphenotype versatility in situ in health and disease.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Recovery Of Intestinal Leukocytes From Lp and Ie Compartmentsmentioning

confidence: 99%

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Modulation of surface CD11c expression tracks plasticity in murine intestinal tissue eosinophils

Larsen

Dockstader

Olbrich

et al. 2022

Journal of Leukocyte Biology

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“…Eosinophil accumulation during inflammatory responses involves their maturation and release from the bone marrow (in approximately 8 days), adhesion, and transmigration from the post-capillary endothelium into peripheral circulation, followed by chemotaxis and activation in tissues. [ 13 ] Many cytokines and chemokines have been shown to mediate this process, most of which are associated with Th2-mediated immune responses. For example, interleukin-3 (IL-3), IL-5, and granulocyte–macrophage colony-stimulating factor (GM-CSF) modulate eosinophil production in the bone marrow, whereas IL-5 is involved in the expansion and release of eosinophils.…”

Section: Pathogenesismentioning

confidence: 99%

Eosinophilic gastroenteritis: Pathogenesis, diagnosis, and treatment

Ruan

Liu

et al. 2023

Chinese Medical Journal

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Eosinophilic gastroenteritis (EGE) is a gastrointestinal disorder of unclear etiology that is characterized by eosinophilic infiltration of the stomach and small intestine, and consists of mucosal, muscular, and serosal subtypes. Eosinophilic infiltration of the gastrointestinal tract is a fundamental histopathological characteristic of EGE and is driven by several T-helper type 2 (Th2)-dependent cytokines and induced by food allergy. Due to the lack of a diagnostic gold standard, EGE has a high rate of delayed diagnosis or misdiagnosis. However, several new diagnostic strategies have been developed, such as novel genetic biomarkers and imaging tests. Although dietary therapy and corticosteroids remain the common choices for EGE treatment, recent decades have seen the emergence of novel treatment alternatives, such as biologics that target particular molecules involved in the pathogenic process. Preliminary investigations and clinical trials have demonstrated the efficacy of biologics and provided additional insights for the era of refractory or corticosteroid-dependent EGE biologics.

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