Segmental allergen challenge in patients with atopic asthma leads to increased IL-9 expression in bronchoalveolar lavage fluid lymphocytes

Erpenbeck, Veit J.; Hohlfeld, Jens M.; Volkmann, Brunhild; Hagenberg, Andreas; Geldmacher, Henning; Braun, Armin; Krug, Norbert

doi:10.1067/mai.2003.1485

Cited by 130 publications

(87 citation statements)

References 25 publications

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“…Reverse transcription was performed with 0.04–0.1 µg of total RNA from each sample in a 40-µl reaction volume for 60 min at 37°C followed by 5 min at 95°C according to the manufacturer’s recommendations (Omniscript RT-Kit, Qiagen, Hilden, Germany). Real-time PCR was performed on the LightCycler Instrument using the DNA binding dye SYBR green (LightCycler-Fast Start DNA Master SYBR Green I, Roche Molecular Biochemicals, Mannheim, Germany), as described previously [26]. PCR reaction mix contained 2 µl of PCR master mix supplemented with 2.5 m M MgCl 2 , custom-synthesized primers (MWG Biotech, Ebersberg, Germany) and 2 µl of either external standards, cDNA or untranscribed RNA to exclude DNA contamination, brought to a final volume of 20 µl with water.…”

Section: Methodsmentioning

confidence: 99%

Regulation of GATA-3, c-maf and T-bet mRNA Expression in Bronchoalveolar Lavage Cells and Bronchial Biopsies after Segmental Allergen Challenge

Erpenbeck¹,

Hagenberg²,

Krentel³

et al. 2006

Int Arch Allergy Immunol

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Background: GATA-3 (GATA binding protein 3) and the proto-oncogene c-maf are Th2-regulating transcription factors that control the expression of interleukin (IL)-4 and IL-5, while T-bet (T-box expressed in T cells) is a Th1-specific transcription factor that controls the expression of interferon (IFN-γ). Allergen provocation in asthmatics induces a Th2-dominated cytokine profile, but so far it is unknown whether the skewed cytokine expression is reflected by the expression of the respective transcriptional regulators. Objective: The aim of this study was to determine the regulation of Th1- and Th2-specific transcription factors and cytokines in 10 atopic subjects with mild asthma and 5 nonatopic healthy controls at baseline and after segmental sham and allergen challenge. Methods: The mRNA expression of GATA-3, c-maf and T-bet was determined by real-time polymerase chain reaction in bronchoalveolar lavage (BAL) cells and bronchial biopsies. The percentage of IL-4+, IL-5+ and IFN-γ+ BAL T cells was determined by flow cytometry, and BAL levels of these cytokines were measured by ELISA. Results: In BAL cells of asthmatics, the mRNA expression of all transcription factors was increased after allergen challenge. In bronchial biopsies, the basal expression of GATA-3 was increased in asthmatics compared to healthy controls but decreased after allergen challenge. Compared to sham challenge, the percentage of IL-5+/CD4+ BAL T cells was increased after allergen challenge in asthmatics while the percentage of IFN-γ+/CD4+ and IFN-γ+/CD8+ T cells was decreased. Expression of c-maf mRNA in BAL cells correlated with IL-4+/CD4+ BAL cells and BAL IL-5 levels. Conclusions: Segmental allergen challenge in asthmatics leads to increased GATA-3, c-maf and T-bet expression in BAL cells but not in bronchial biopsies.

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Section: Methodsmentioning

confidence: 99%

Regulation of GATA-3, c-maf and T-bet mRNA Expression in Bronchoalveolar Lavage Cells and Bronchial Biopsies after Segmental Allergen Challenge

Erpenbeck¹,

Hagenberg²,

Krentel³

et al. 2006

Int Arch Allergy Immunol

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“…IL-9, a pleiotropic cytokine, has been reported to have an important role in atopic asthma (22,23). Studies have found that naïve CD8 + T cells are also able to differentiate into Tc9 cells under similar Th9-polarizing conditions (12,24).…”

Section: Cd8mentioning

confidence: 99%

Increased circulating IL-9-producing CD8+ T cells are associated with eosinophilia and high FeNO in allergic asthmatics

Wang

Cheng

Chen

et al. 2016

Experimental and Therapeutic Medicine

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Abstract. Allergic asthma is a chronic airway disorder mediated by Th2 cells. It has been shown that IL-9-producing CD8 + cytotoxic T (Tc9) cells promote the subsequent onset of allergic airway inflammation in mice mediated by abnormal Th2 immunity. Whether Tc9 cells are associated with the immunopathogenesis of asthmatic patients remains unknown. In the present study, peripheral blood mononuclear cells (PBMCs) were separated by Ficoll-Hypaque gradient centrifugation from all subjects. The frequency of Tc9 cells was measured by flow cytometry. Serum IL-9 levels were assessed by enzyme-linked immunosorbent assay (ELISA). mRNA expression levels of IL-9, STAT6, and IRF4 in PBMCs from healthy controls and asthmatic patients were detected by reverse transcription-quantitative polymerase chain reaction. The results showed that the numbers of Tc9 cells in allergic asthmatics were significantly increased, compared with healthy controls (P<0.0001). Notably, IL-9 protein and mRNA levels were increased in allergic asthmatics and STAT6 and IRF4 mRNA levels were elevated, as compared with healthy controls. In addition, circulating numbers of Tc9 cells were positively correlated with blood eosinophil counts and fractioned exhaled nitric oxide (FeNO) levels in asthmatic patients. Moreover, the number of Tc9 cells and serum IL-9 levels in asthmatic patients were significantly decreased after treatment with glucocorticoids (P<0.05). These findings suggest that increased circulating Tc9 cells are associated with eosinophilia and high FeNO of allergic asthma, and that abnormal Tc9 immunity may contribute to the pathogenesis of allergic asthmatics.

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“…Cells were thawed, cultured up to passage 3, harvested and stored frozen until RNA isolation and cDNA transcription. Primers (Müller et al 2006) and protocols (Erpenbeck et al 2003) were those used previously. Abundance of mRNA was determined via the ratio of target to house-keeping gene (PBGD, porphobilinogen deaminase).…”

Section: Gene Expression Analysismentioning

confidence: 99%

In contrast to lung fibroblasts – no signs of senescence in skin fibroblasts of patients with emphysema

Müller

Paasch

Feindt

et al. 2008

Experimental Gerontology

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Segmental allergen challenge in patients with atopic asthma leads to increased IL-9 expression in bronchoalveolar lavage fluid lymphocytes

Cited by 130 publications

References 25 publications

Regulation of GATA-3, c-maf and T-bet mRNA Expression in Bronchoalveolar Lavage Cells and Bronchial Biopsies after Segmental Allergen Challenge

Regulation of GATA-3, c-maf and T-bet mRNA Expression in Bronchoalveolar Lavage Cells and Bronchial Biopsies after Segmental Allergen Challenge

Increased circulating IL-9-producing CD8+ T cells are associated with eosinophilia and high FeNO in allergic asthmatics

In contrast to lung fibroblasts – no signs of senescence in skin fibroblasts of patients with emphysema

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