Detection of True IgE-expressing Mouse B Lineage Cells

Gallagher, M. J.; Shrestha, Akritee; Magee, Jennifer; Wesemann, Duane R.

doi:10.3791/52264

Cited by 3 publications

(3 citation statements)

References 21 publications

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“…Cells were counted using a hemocytometer with exclusion of dead cells with Trypan blue dye. Cells were stained with fluorophore or biotinconjugated antibodies as described (54,55) where indicated. The cell sorting was performed on a FACSAria II flow cytometer (BD Biosciences).…”

Section: Methodsmentioning

confidence: 99%

“…The cell sorting was performed on a FACSAria II flow cytometer (BD Biosciences). For the in vitro CSR experiments and the allelic exclusion experiments, staining for IgM, IgG1, and IgE for intracytoplasmic IgH expression was done by using trypsinization followed by fixation/permeabilization as described (54,55). Data analysis was performed with FlowJo software (v9.9.4).…”

Section: Methodsmentioning

confidence: 99%

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IgH isotype-specific B cell receptor expression influences B cell fate

Tong

Granato

Zuo

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Ig heavy chain (IgH) isotypes (e.g., IgM, IgG, and IgE) are generated as secreted/soluble antibodies (sIg) or as membrane-bound (mIg) B cell receptors (BCRs) through alternative RNA splicing. IgH isotype dictates soluble antibody function, but how mIg isotype influences B cell behavior is not well defined. We examined IgH isotype-specific BCR function by analyzing naturally switched B cells from wild-type mice, as well as by engineering polyclonal γ1/γ1 andε/ε mice, which initially produce IgG1 or IgE from their respective native genomic configurations. We found that B cells from wild-type mice, as well as γ1/γ1 andε/ε mice, produce transcripts that generate IgM, IgG1, and IgE in an alternative splice form bias hierarchy, regardless of cell stage. In this regard, we found that mIgμ > mIgγ1 > mIgε, and that these BCR expression differences influence respective developmental fitness. Restrained B cell development from γ1/γ1 andε/ε mice was proportional to sIg/mIg ratios and was rescued by enforced expression of the respective mIgs. In addition, artificially enhancing BCR signal strength permitted IgE memory B cells-which essentially do not exist under normal conditions-to provide long-lived memory function, suggesting that quantitative BCR signal weakness contributes to restraint of IgE B cell responses. Our results indicate that IgH isotype-specific mIg/BCR dosage may play a larger role in B cell fate than previously anticipated.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

IgH isotype-specific B cell receptor expression influences B cell fate

Tong

Granato

Zuo

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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“…After staining, cells were washed twice with BD Biosciences Perm/Wash buffer. IgE staining was performed using the methods described by Gallagher et al with anti-IgE–FITC (1:100, BioLegend 406906) ( 60 ).…”

Section: Methodsmentioning

confidence: 99%

Nociceptor neurons promote IgE class switch in B cells

Mathur¹,

Wang²,

Seehus³

et al. 2021

JCI Insight

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Nociceptors, the high-threshold primary sensory neurons that trigger pain, interact with immune cells in the periphery to modulate innate immune responses. Whether they also participate in adaptive and humoral immunity is, however, not known. In this study, we probed if nociceptors have a role in distinct airway and skin models of allergic inflammation. In both models, the genetic ablation and pharmacological silencing of nociceptors substantially reduced inflammatory cell infiltration to the affected tissue. Moreover, we also found a profound and specific deficit in IgE production in these models of allergic inflammation. Mechanistically, we discovered that the nociceptor-released neuropeptide substance P helped trigger the formation of antibody-secreting cells and their release of IgE. Our findings suggest that nociceptors, in addition to their contributions to innate immunity, play a key role in modulating the adaptive immune response, particularly B cell antibody class switching to IgE.

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Inhibition of immunoglobulin E attenuates pulmonary hypertension

et al. 2022

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Pulmonary hypertension (PH) is a severe cardiopulmonary disease characterized by pathological vascular remodeling in the lung. Immunoglobulin E (IgE) is known to participate in aortic vascular remodeling, but whether IgE mediates pulmonary vascular remodeling in PH is unknown. Here, we found serum IgE elevation in PAH patients, hypoxia-induced PH mice and monocrotaline (MCT)-induced PH rats.Combining animal model of PH with single-cell RNA sequencing, we found IgE production in the lung tissues of PH mice. Neutralizing IgE with an anti-IgE antibody was effective in preventing PH development in mice and rat models. The IgE receptor FcεRIα was also upregulated in PH lung tissues and Fcer1a de ciency prevented the development of PH in mice. Single-cell RNA-seq revealed that FcεRIα was mostly expressed in mast cells, and mast cell-speci c Fcer1a knockout protected against PH in mice.Further mechanistic experiments revealed that IgE-activated mast cells produced interleukins IL6 and IL13, which subsequently promoted vascular muscularization. Clinically approved IgE antibody Omalizumab alleviated the progression of established PH in rats. Using genetic and pharmacological approaches, we have demonstrated that blocking IgE-FcεRIα signaling may hold potential for the treatment of PAH.

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Detection of True IgE-expressing Mouse B Lineage Cells

Cited by 3 publications

References 21 publications

IgH isotype-specific B cell receptor expression influences B cell fate

IgH isotype-specific B cell receptor expression influences B cell fate

Nociceptor neurons promote IgE class switch in B cells

Inhibition of immunoglobulin E attenuates pulmonary hypertension

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