Antigen profiles for the quantitative assessment of eosinophils in mouse tissues by flow cytometry

Dyer, Kimberly D.; Garcia-Crespo, Katia E.; Killoran, Kristin E.; Rosenberg, Helene F.

doi:10.1016/j.jim.2011.04.009

Cited by 43 publications

(42 citation statements)

References 24 publications

(27 reference statements)

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“…Siglec-F was also found to be expressed by AMs, a finding consistent with that of previous studies (28,29), and by lung neutrophils, which has not been reported before. Although CD125 expression is thought to be restricted to eosinophils and B-1 cells in mice (28,29,(62)(63)(64), we unexpectedly found that lung neutrophils expressed high levels of CD125, suggesting that these cells may also be influenced by IL-5. Supporting this finding, mouse neutrophils have been shown to migrate in response to rIL-5 treatment (65).…”

Section: Discussionsupporting

confidence: 92%

Lung-resident eosinophils represent a distinct regulatory eosinophil subset

Mesnil

Raulier

Paulissen

et al. 2016

Journal of Clinical Investigation

431

506

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Increases in eosinophil numbers are associated with infection and allergic diseases, including asthma, but there is also evidence that eosinophils contribute to homeostatic immune processes. In mice, the normal lung contains resident eosinophils (rEos), but their function has not been characterized. Here, we have reported that steady-state pulmonary rEos are IL-5-independent parenchymal Siglec-FintCD62L+CD101lo cells with a ring-shaped nucleus. During house dust mite-induced airway allergy, rEos features remained unchanged, and rEos were accompanied by recruited inflammatory eosinophils (iEos), which were defined as IL-5-dependent peribronchial Siglec-FhiCD62L-CD101hi cells with a segmented nucleus. Gene expression analyses revealed a more regulatory profile for rEos than for iEos, and correspondingly, mice lacking lung rEos showed an increase in Th2 cell responses to inhaled allergens. Such elevation of Th2 responses was linked to the ability of rEos, but not iEos, to inhibit the maturation, and therefore the pro-Th2 function, of allergen-loaded DCs. Finally, we determined that the parenchymal rEos found in nonasthmatic human lungs (Siglec-8+CD62L+IL-3Rlo cells) were phenotypically distinct from the iEos isolated from the sputa of eosinophilic asthmatic patients (Siglec-8+CD62LloIL-3Rhi cells), suggesting that our findings in mice are relevant to humans. In conclusion, our data define lung rEos as a distinct eosinophil subset with key homeostatic functions

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Section: Discussionsupporting

confidence: 92%

Lung-resident eosinophils represent a distinct regulatory eosinophil subset

Mesnil

Raulier

Paulissen

et al. 2016

Journal of Clinical Investigation

431

506

View full text Add to dashboard Cite

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“…3A, 3B). Interestingly, strict gating based on the FMO controls incorporated a range of Siglec-F expressing cells and higher overall percentages than have been previously reported for mature eosinophils (25). To further examine eosinophil related populations within this Siglec-F + pool, we defined a population of GMP-like cells (Lin − Sca1 − Siglec-F + IL-5Rα − SSC lo ckit hi CD34 + ST2 + ), eosinophil precursors (EoPre) (Lin hi Sca1 − Siglec-F lo IL-5Rα + SSC lo ckit − CD34 − ST2 − ), and EoM cells (Lin lo Sca1 − Siglec-F hi IL-5Rα + SSC hi ckit − CD34 − ST2 lo ) (characterization and FMO control staining shown in Supplemental Fig.…”

Section: Resultsmentioning

confidence: 81%

IL-33 Precedes IL-5 in Regulating Eosinophil Commitment and Is Required for Eosinophil Homeostasis

Johnston

Hsu

Krier-Burris

et al. 2016

The Journal of Immunology

130

109

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Eosinophils are important in the pathogenesis of many diseases, including asthma, eosinophilic esophagitis, and eczema. While IL-5 is crucial for supporting mature eosinophils, the signals that support earlier eosinophil lineage events are less defined. The IL-33 receptor, ST2, is expressed on several inflammatory cells, including eosinophils, and is best characterized for its role during the initiation of allergic responses in peripheral tissues. Recently, ST2 expression was described on hematopoietic progenitor subsets, where its function remains controversial. Our findings demonstrate that IL-33 is required for basal eosinophil homeostasis, since both IL-33– and ST2-deficient mice exhibited diminished peripheral blood eosinophil numbers at baseline. Exogenous IL-33 administration increased mature eosinophils in both the bone marrow and periphery in WT and IL-33–deficient, but not ST2-deficient, mice. Systemic IL-5 was also increased under this treatment, and blocking IL-5 with a neutralizing antibody ablated of the IL-33-induced mature eosinophil expansion. The homeostatic hypereosinophilia seen in IL-5–transgenic mice was significantly lower with ST2 deficiency despite similar elevations in systemic IL-5. Finally, in vitro treatment of bone marrow cells with IL-33, but not IL-5, led to specific early expansion of IL-5Rα–expressing precursor cells. In summary, our findings establish a basal defect in eosinophilopoiesis in IL-33– and ST2-deficient mice and a mechanism whereby IL-33 supports mature eosinophils by driving both systemic IL-5 production and the expansion of IL-5Rα–expressing precursor cells.

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“…Spleen cell suspensions in HBSS supplemented with 1% FBS and 10 mM HEPES were prepared as described (Dyer et al , 2011). Red blood cells were lysed with ACK lysing buffer (Lonza).…”

Section: Methodsmentioning

confidence: 99%

Signaling via pattern recognition receptors NOD2 and TLR2 contributes to immunomodulatory control of lethal pneumovirus infection

et al. 2016

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Pattern recognition receptors (PRRs) engage microbial components in the lung, although their role in providing primary host defense against respiratory virus infection is not fully understood. We have previously shown that Gram-positive Lactobacillus plantarum (Lp) administered to the respiratory tract promotes full and sustained protection in response to an otherwise lethal mouse pneumovirus (PVM) infection, a robust example of heterologous immunity. While Lp engages PRRs TLR2 and NOD2 in ex vivo signaling assays, we found that Lp-mediated protection was unimpaired in single gene-deleted TLR2−/− and NOD2−/− mice. Here we demonstrate substantial loss of Lp-mediated protection in a double gene-deleted NOD2−/−TLR2−/− strain. Furthermore, we demonstrate protection against PVM infection by administration of the bi-functional NOD2-TLR2 agonist, CL-429. The bi-functional NOD2-TLR2 ligand CL-429 not only suppresses virus-induced inflammation, it is significantly more effective at preventing lethal infection than equivalent amounts of mono-molecular TLR2 and NOD2 agonists. Interestingly, and in contrast to biochemical NOD2 and/or TLR2 agonists, Lp remained capable of eliciting primary proinflammatory responses from NOD2−/−TLR2−/− mice in vivo and from alveolar macrophages challenged ex vivo. Taken together, we conclude that coordinate engagement of NOD2 and TLR2 constitutes a key step in the genesis of Lp-mediated protection from a lethal respiratory virus infection, and represents a critical target for modulation of virus-induced inflammatory pathology.

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Antigen profiles for the quantitative assessment of eosinophils in mouse tissues by flow cytometry

Cited by 43 publications

References 24 publications

Lung-resident eosinophils represent a distinct regulatory eosinophil subset

Lung-resident eosinophils represent a distinct regulatory eosinophil subset

IL-33 Precedes IL-5 in Regulating Eosinophil Commitment and Is Required for Eosinophil Homeostasis

Signaling via pattern recognition receptors NOD2 and TLR2 contributes to immunomodulatory control of lethal pneumovirus infection

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