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. 2017 Jul 19;18(1):38.
doi: 10.1186/s12865-017-0220-1.

Detection of IL-9 producing T cells in the PBMCs of allergic asthmatic patients

Lei Jia  1 , Ying Wang  2 , Jiangping Li  1 , Sha Li  3 , Yannan Zhang  1 , Juan Shen  1 , Weiping Tan  4 , Changyou Wu  5
Affiliations

Detection of IL-9 producing T cells in the PBMCs of allergic asthmatic patients

Lei Jia et al. BMC Immunol. .

Abstract

Background: Interleukin-9 (IL-9) was reported as an active participant in the pathogenesis of allergic asthma. This study aimed to investigate the major source ofIL-9 and its effect on interferon γ (IFN-γ) and immunoglobulin (Ig) secretion by B cells.

Methods: We isolated peripheral blood mononuclear cells from children with allergic asthma and healthy children. IL-9, IL-4 and IFN-γ expression were detected by ELISA, ELISpot and Flowcytometry. Expression of transcription factor PU.1 was measured by Western Blot. We evaluated the effect of IL-9 on B cell activation and Ig production.

Results: Results showed that compared with healthy children, levels of IL-9, IL-4 and PU.1 were elevated and levels of IFN-γ were lower in children with allergic asthma. IL-9-expressing CD4+ T cells did not co-express IL-4. Exogenous IL-9 inhibited IFN-γ production in a dose-dependent manner. Antigen-specific Th9 cells existed in children with house dust mite allergic asthma. IL-9 up-regulated expression of CD69 and CD25 on B cells and combination of IL-9 and IL-4 enhanced IgE production.

Conclusions: In conclusion, our results showed that Th9 cells may be the major source of IL-9 in children with allergic asthma. In these patients, IL-9 impairs IFN-γ production and synergistically promotes IL-4-induced IgE secretion.

Keywords: Allergic asthma; IL-9; IgE; Th9 cell.

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Conflict of interest statement

Ethics approval and consent to participate

The experimental protocols were established following the Declaration of Helsinki and approved by the Ethical Committee of Sun Yat-sen Memorial Hospital and Zhongshan School of Medicine, Sun Yat-sen University. Written informed consent was obtained from the legal guardians of all participants.

Consent for publication

Not applicable.

Competing interests

The authors declare no financial or commercial conflict of interest.

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Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Figures

Fig. 1
Fig. 1
IL-9, IL-4 and IFN-γ production by PBMCs from children with allergic asthma. IL-9 (a), IL-4 (c) and IFN-γ (e) production by PBMCs from PBMCs of healthy donors (HD) and allergic asthma patients (AP) was detected by ELISA. Representative and statistical ELISpot data of IL-9 (b), IL-4 (d) and IFN-γ (f) were obtained from 3 independent experiments. ELISA data were collected from 10 healthy donors and 14 asthma patients. *, P < 0.05; **, P < 0.01; ***, P < 0.001
Fig. 2
Fig. 2
IL-9 inhibits the production of IFN-γ. IL-9 inhibits IFN-γ production from PBMCs in a dose-dependent manner (a). IFN-γ production by purified CD4+ T cells was detected by ELISA (b). ELISA data were collected from 3 independent experiments. ***,P < 0.001
Fig. 3
Fig. 3
The major IL-9-expressing CD4+ T cells in children with allergic asthma are Th9 cells. The percentage of Th9 cells in PBMCs of healthy donors and asthma patients was detected by FACS (a). Statistical result was obtained from HD (n = 10) and AP (n = 12) (b). PU.1 levels in total HD and AP PBMCs were measured by Western Blot (c). **, P < 0.01
Fig. 4
Fig. 4
HDM-specific Th9 cells in children with allergic asthma. a The production of IL-9 by PBMCs of children with house dust mite allergic asthma (n = 5). PBMCs were stimulated with house dust mite (HDM) extract and IL-9 was detected by ELISA. b Representative FACS data of HDM-specific Th9 cell in children with HDM allergic asthma. c Statistical result of HDM-specific Th9 cell in children with HDM allergic asthma (n = 3)
Fig. 5
Fig. 5
IL-9 promotes B cells activation. a Representative FACS data of IL-9 activation function on B cells. b Statistical results of IL-9 activation function on B cells. IL-9, alone, up-regulates the Mean Fluorescence Intensity (MFI) of CD69 and CD25 and increases the percentage of CD25 (n = 6). c Representative FACS data of synergistic effect of IL-9 on B cell activation. d Statistical results of IL-9 synergistic activation function on B cells. IL-9, synergistically with Anti-CD40, up-regulates the MFI of CD69 and increases the percentage of CD25 (n = 10). ns, P > 0.05; *, P < 0.05; **, P < 0.01; ***, P < 0.001
Fig. 6
Fig. 6
IL-9 synergistically promotes IL-4-induced IgE and IgG production. a The production of IgG, but not IgE, can be promoted by IL-9 alone (n = 3). b Synergistic effect of IL-9 on IL-4–induced IgE and IgG production (n = 4). IgE and IgG levels were detected by ELISA. Results represent the Mean ± SEM. ND, not detected; **, P < 0.01; ***, P < 0.001
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