Measuring mast cell degranulation

Berger, Stuart A.

doi:10.1016/b978-012442710-5.50154-4

Cited by 3 publications

(3 citation statements)

References 10 publications

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“…32 Briefly, BMMCs were loaded with 5 g/mL dinitrophenyl (DNP)-specific IgE (SPE-1, Sigma) for 30 minutes, washed, and resuspended in Tyrode buffer (10 mM HEPES [N-2-hydroxyethylpiperazine-NЈ-2-ethanesulfonic acid] buffer at pH 7.4, 130 mM NaCl, 5 mM KCl, 1.4 mM CaCl 2 , 1 mM MgCl 2 , 5.6 mM glucose and 0.1% bovine serum albumin [BSA]). Degranulation was induced by stimulation with DNP-human serum albumin (HSA), PMA, and ionomycin.…”

Section: Degranulation Assaymentioning

confidence: 99%

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TLR3-induced activation of mast cells modulates CD8+ T-cell recruitment

Orinska

Bulanova

Budagian

et al. 2005

Blood

156

162

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IntroductionThe magnitude and quality of innate immune response is essential for appropriate adaptive response. The activation of naive CD8 ϩ T cells, the main effector cells in the course of viral infections, their clonal expansion, development of effector cells, and maintenance and expansion of memory CD8 ϩ T cells after antigen reappearance are precisely regulated and dependent on an adequate context of costimulation and cytokine/chemokine environment, provided by cells responding to "first defense line" signals of innate immunity. [1][2][3][4] Mast cells (MCs), pivotal effector cells in immunoglobulin (Ig) E-associated disorders, have recently been recognized as important elements in innate immune defenses. 5 Strategically located at host/environment interfaces like the skin, airways, and gastrointestinal and urogenital tracts, 6,7 MCs are equipped with a large variety of receptors to detect signs of infections including Toll-like receptors (TLRs), CD48, and complement receptors. 8 MCs activated via these receptors secrete a large number of proinflammatory products including granule-associated preformed mediators (histamine, serotonin, proteases, proteoglycans), lipid mediators (leukotrienes B 4 and C 4 and prostaglandins), as well as cytokines, chemokines, and growth factors. 9 In contrast to the well-established crucial role of MCs in induction of host defense responses to bacteria and parasites, the function of MCs in antiviral immune responses remains to be characterized in detail. Recently, a number of reports have shown the functional interplay between MCs and T cells in allergic, infectious, and autoimmune processes, for example, MCs and T cells colocalizing in inflamed tissues, as in the lungs of patients with asthma and in animal models of allergic asthma. 10,11 It has also been reported that tumor necrosis factor ␣ (TNF-␣) released from MCs contributes substantially to T-cell recruitment to the draining lymph nodes (LNs) in an experimental infection model with Escherichia coli. 12 Furthermore, activated MCs induce the chemotaxis of CD8 ϩ T cells through the production of leukotrienes in vitro. 13 Further, the recruitment of early effector T cells into the airways in an asthma model is mediated by leukotriene B 4 (LTB 4 ) 14 released by MCs activated by IgE cross-linking. 15 However, the mechanisms and relevance of MC-T-cell interactions, as well as the signals required for MC activation and effector functions in settings of viral infections remain largely unknown.TLRs play an important role in innate immunity recognizing specific, nonself, conserved components shared by different pathogens. Different TLRs are involved in the specific recognition of various microbes. 16,17 Human and mouse MCs have been shown to be activated by bacterial peptidoglycan and lipopolysaccharide (LPS) via TLR2 and TLR4, respectively, producing a number of cytokines such as interleukin 5 (IL-5), IL-6, IL-10, IL-13, and TNF-␣. [18][19][20][21] On binding of viral dsRNA or its synthetic counterpart polyinosinic-polycytidyl...

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Section: Degranulation Assaymentioning

confidence: 99%

“…32 Briefly, BMMCs were loaded with 5 g/mL dinitrophenyl (DNP)-specific IgE (SPE-1, Sigma) for 30 minutes, washed, and resuspended in Tyrode buffer (10 mM For personal use only. on May 9, 2018. by guest www.bloodjournal.org From absorbance in the supernatants by the sum of absorbance in the supernatant and cell lysate.…”

Section: Degranulation Assaymentioning

confidence: 99%

TLR3-induced activation of mast cells modulates CD8+ T-cell recruitment

Orinska

Bulanova

Budagian

et al. 2005

Blood

156

162

View full text Add to dashboard Cite

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“…To analyze BMMC proliferation in response to IL-3, 2 ϫ 10 6 cells/mL were incubated with IL-3 for 72 hours, and 1 Ci 3 H-thymidine was added for the last 18 hours. Measurement of histamine by o-phthaldialdehyde assay, 22 measurement of MC degranulation by ␤-hexosaminidase release, 23 RNA extraction, cDNA synthesis, and conventional semiquantitative reverse transcription polymerase chain reaction (RT-PCR) for MC proteases are described in the supplemental data (available on the Blood Web site; see the Supplemental Materials link at the top of the online article).…”

Section: Cell Isolation Generation Culture and Stimulationmentioning

confidence: 99%

I787 provides signals for c-Kit receptor internalization and functionality that control mast cell survival and development

Orinska

Föger

Huber

et al. 2010

Blood

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ATP induces P2X7 receptor-independent cytokine and chemokine expression through P2X1 and P2X3 receptors in murine mast cells

Bulanova

Budagian

Orinska

et al. 2009

Journal of Leukocyte Biology

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Extracellular ATP mediates a diverse array of biological responses in many cell types and tissues, including immune cells. We have demonstrated that ATP induces purinergic receptor P2X(7) mediated membrane permeabilization, apoptosis, and cytokine expression in murine mast cells (MCs). Here, we report that MCs deficient in the expression of the P2X(7) receptor are resistant to the ATP-induced membrane permeabilization and apoptosis. However, ATP affects the tyrosine phosphorylation pattern of P2X(7)knockout cells, leading to the activation of ERK1/2. Furthermore, ATP induces expression of several cytokines and chemokines in these cells, including IL-4, IL-6, IFN-gamma, TNF-alpha, RANTES, and MIP-2, at the mRNA level. In addition, the release of IL-6 and IL-13 to cell-conditioned medium was confirmed by ELISA. The ligand selectivity and pharmacological profile indicate the involvement of two P2X family receptors, P2X(1) and P2X(3). Thus, depending on genetic background, particular tissue microenvironment, and ATP concentration, MCs can presumably engage different P2X receptor subtypes, which may result in functionally distinct biological responses to extracellular nucleotides. This finding highlights a novel level of complexity in the sophisticated biology of MCs and may facilitate the development of new therapeutic approaches to modulate MC activities.

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Measuring mast cell degranulation

Cited by 3 publications

References 10 publications

TLR3-induced activation of mast cells modulates CD8+ T-cell recruitment

TLR3-induced activation of mast cells modulates CD8+ T-cell recruitment

I787 provides signals for c-Kit receptor internalization and functionality that control mast cell survival and development

ATP induces P2X7 receptor-independent cytokine and chemokine expression through P2X1 and P2X3 receptors in murine mast cells

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