Use of the relative release index for histamine in LAD2 cells to evaluate the potential anaphylactoid effects of drugs

Han, Shengli; Lv, Yanni; Kong, Liyun; Che, Delu; Liu, Rui; Jia, Fu; Cao, Jiao; Wang, Jue; Wang, Cheng; He, Huaizhen; Zhang, Tao; Dong, Xinzhong; He, Langchong

doi:10.1038/s41598-017-14224-z

Cited by 40 publications

(23 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It has been shown in human LAD2 mast cell line that ciprofloxacin induced calcium influx . In our hands, in both granulocyte subtypes, direct stimulation with ciprofloxacin significantly and dose‐dependently induced calcium influx that could be significantly blocked by prior blocking with anti‐MRGPRX2 mAb indicating that ciprofloxacin might act through MRGPRX2 receptor.…”

Section: Discussionsupporting

confidence: 50%

The pseudoallergen receptor MRGPRX2 on peripheral blood basophils and eosinophils: Expression and function

Wedi

Gehring

Kapp

2020

Allergy

View full text Add to dashboard Cite

Background Mas‐related G protein‐coupled receptor X2 (MRGPRX2) is regarded as a mast cell‐specific receptor mediating non–IgE‐dependent activation. We aimed to investigate whether human basophils and eosinophils express functional MRGPRX2. Methods Flow cytometry, immunocytochemistry, immunofluorescence, Western blot, and RT‐PCR were performed in highly purified peripheral blood basophils and eosinophils of atopic and nonatopic donors. To assess functional activity, fluorescent avidin‐based degranulation assay, calcium mobilization, cytokine production in supernatants, assessment of viability/apoptosis, and tricolor granulocyte activation test were used. Results MRGPRX2 was significantly expressed by basophils and eosinophils but not neutrophils. Functional capacity was shown by anti‐MRGPRX2 mAb‐induced calcium influx and concentration‐dependent induction of degranulation. Sequential stimulation in the calcium mobilization assay gave no evidence for desensitization or receptor internalization. Anti‐MRGPRX2 mAb significantly promoted survival. Inhibition of apoptosis could be due to released IL‐3, IL‐5, and GM‐CSF found in supernatants. Short‐term incubation with IL‐3 dose‐dependently upregulated MRGPRX2 expression in both, stimulation for 24 hours with anti‐IgE, C5a, fMLP, and IL‐3 in basophils and by IL‐3, IL‐5, and IL‐33 in eosinophils. Among known mast cell MRGPRX2 agonists ciprofloxacin but not PMX‐53 was functional on basophils and eosinophils. In basophils of allergic subjects, tricolor granulocyte activation test using grass pollen demonstrated MRGPRX2 upregulation associated with degranulation and CD63 expression. Conclusion Unraveling the regulation and signaling mechanisms of MRGPRX2 on basophils and eosinophils might enable the development of new therapeutic strategies to prevent or inhibit allergic and nonallergic hypersensitivity. Moreover, addressing MRGPRX2 might have potential for diagnostic purposes in (drug) hypersensitivity.

show abstract

Section: Discussionsupporting

confidence: 50%

The pseudoallergen receptor MRGPRX2 on peripheral blood basophils and eosinophils: Expression and function

Wedi

Gehring

Kapp

2020

Allergy

View full text Add to dashboard Cite

show abstract

“…Fifty microliter of supernatant was collected. Histamine release assay was carried out using LC‐ESI‐MS/MS method (Han et al, 2017). In the applied LC‐ESI‐MS/MS method, an LCMS 8040 mass spectrometer (Shimadzu Corporation, Kyoto, Japan) was used.…”

Section: Methodsmentioning

confidence: 99%

Isoimperatorin reduces the effective dose of dexamethasone in a murine model of asthma by inhibiting mast cell activation

Wang

Zhang

Zeng

et al. 2020

Phytotherapy Research

Self Cite

View full text Add to dashboard Cite

Adverse effects that result from dexamethasone (DEX) use are common and serious in patients with asthma. Therefore, alternative anti-inflammatory treatments are being investigated. Isoimperatorin (ISO), an active natural furocoumarin, possesses multiple pharmacological properties, including an anti-inflammation effect. In this study, investigations were conducted on the effect of ISO on mast cell (MC) activation in vitro and whether ISO could reduce the effective dose of DEX in a mast cell-dependent murine model of asthma in vivo. Calcium imaging was used to assess intracellular Ca 2+ mobilization. Enzyme-linked immunosorbent assay was used to measure the chemokines release. Western blot analysis was conducted to investigate the underlying pathway. Airway inflammation and hyperresponsiveness (AHR) were examined in an asthma model. ISO inhibited Ca 2+ flux and MC degranulation via Lyn/PLCγ1/PKC, ERK, and P38 MAPK pathways. In the asthma model, ISO, in combination with DEX, showed an additive inhibitory effect on AHR, inflammation, and the number of activated MCs in the lungs and decreased the levels of interleukin (IL)-4, IL-5, IL-6, IL-13, tumor necrosis factor (TNF)-a, and CC motif chemokine ligand (CCL)-2 in bronchoalveolar lavage fluid. A combination of DEX and ISO may be appropriate if a decrease in the steroid dose is desired owing to dose-dependent adverse effects.

show abstract

“…The samples were separated using a Venusil HILIC column (150 × 2.1 mm id, 3 µm, Agela Technologies, Tianjin, China) and a mobile phase consisting of acetonitrile and ammonium formate (20 mM)–0.1% formic acid aqueous solution (80:20, v/v) at a flow rate of 0.3 mL/min. Other optimized parameter conditions were maintained according to our previous work . Data are expressed as the mean ± standard error of the mean (SEM).…”

Section: Methodsmentioning

confidence: 99%

Screening the anti‐allergic components in Saposhnikoviae Radix using high‐expression Mas‐related G protein‐coupled receptor X2 cell membrane chromatography online coupled with liquid chromatography and mass spectrometry

Jia

Wei

Zhang

et al. 2019

J of Separation Science

Self Cite

View full text Add to dashboard Cite

Saposhnikoviae Radix, the dried root of Saposhnikoviae divaricata, is commonly used in the traditional Chinese anti-allergic preparations, like Bofutsusho-san and Yupingfeng granules. A high-expression Mas-related G protein-coupled receptor X2 cell membrane chromatography coupled online with high-performance liquid chromatography combined with an ion trap time-of-flight multistage mass spectrometry system was established and used for screening and identifying the anti-allergic components in Saposhnikoviae Radix. The system was validated for excellent specificity and suitability using the appropriate standards. Two retained fractions were obtained on the cell membrane chromatography column, and three main components were identified as prim-O-glucosylcimifugin, cimifugin, and 4 ′ -O-β-D-glucosyl-5-Omethylvisamminol. Next, the molecular docking study was conducted, which confirmed that these three components could effectively bind to MRGPRX2 through hydrogen bonds with its amino acid residues. Finally, histamine release assay was performed to investigate the bioactivities of prim-O-glucosylcimifugin, cimifugin, and 4 ′ -O-β-D-glucosyl-5-O-methylvisamminol. Results showed that these three components could exert anti-allergic effects by inhibiting the histamine release in a dosedependent manner (from 10 to 100 μM). In conclusion, the high-expression Masrelated G protein-coupled receptor X2 cell membrane chromatography is an effective tool for discovering the anti-allergic components in Saposhnikoviae Radix. K E Y W O R D Sallergy, cell membrane chromatography, multistage mass spectrometry, traditional Chinese medicine Article Related Abbreviations: CF, cimifugin; CFH, ciprofloxacin hydrochloride; CMC, cell membrane chromatography; MRGPRX2, Mas-related G protein-coupled receptor X2; MV, 4 ′ -O-β-D-glucosyl-5-O-methylvisamminol; PGF, prim-O-glucosylcimifugin; SME, the methanol extract of Saposhnikoviae Radix.

show abstract

Use of the relative release index for histamine in LAD2 cells to evaluate the potential anaphylactoid effects of drugs

Cited by 40 publications

References 20 publications

The pseudoallergen receptor MRGPRX2 on peripheral blood basophils and eosinophils: Expression and function

The pseudoallergen receptor MRGPRX2 on peripheral blood basophils and eosinophils: Expression and function

Isoimperatorin reduces the effective dose of dexamethasone in a murine model of asthma by inhibiting mast cell activation

Screening the anti‐allergic components in Saposhnikoviae Radix using high‐expression Mas‐related G protein‐coupled receptor X2 cell membrane chromatography online coupled with liquid chromatography and mass spectrometry

Contact Info

Product

Resources

About