Th-17 regulatory cytokines promote interleukins-17A and 17F production by neutrophils during asthma

doi:10.3969/j.issn.2095-4344.2114

Abstract

Abstract:

BACKGROUND: Th-17 cells and their derived cytokines (interleukins 17, 21, and 22) show an increasing trend in patients with severe asthma, but the specific regulatory mechanism has not yet been determined.

OBJECTIVE: To investigate the effect of Th-17 regulatory cytokines on the expression of interleukin 17 in neutrophils in patients with severe asthma.

METHODS: There were 28 patients with asthma and 28 healthy controls in the study. Peripheral blood neutrophils isolated from all the subjects were stimulated with interleukins 21, 23, and 6 cytokines and their ability to produce interleukin 17A and 17F was determined relative to healthy controls. Signal transducer and activator of transcription 3 (STAT3) phosphorylation levels were measured in stimulated neutrophils using ﬂow cytometry. The requirement for STAT3 phosphorylation was determined by blocking its activation using a specific chemical inhibitor. The study protocol was reviewed and approved by the Ethics Committee of the Affiliated Hospital of Guilin Medical University, with an approval No. 2017-013.

RESULTS AND CONCLUSION: Stimulating asthmatic neutrophils with interleukins 21, 23, and 6 significantly enhanced the production of interleukins-17A and 17F in neutrophils, and increased STAT3 phosphorylation in all the patients compared with the healthy controls. Interestingly, inhibiting STAT3 phosphorylation using a specific chemical inhibitor dramatically blocked the ability of neutrophils to produce interleukin 17, demonstrating that STAT3 activation is the major approach to mediate the expression of interleukin 17 gene. To conclude, neutrophil infiltration in lungs of severe asthmatics may represent an important source of pro-inﬂammatory interleukins 17A and 17F. STAT3 pathway may be a potential target for regulating neutrophilic inﬂammation during severe asthma.

Key words: asthma, cytokines, inflammatory response, neutrophils, Th-17 cells, STAT3

CLC Number:

Wei Jianghong, Jia Aijun, Ma Libing, Wang Yueling, Qiu Lulu, Xiao Bing. Th-17 regulatory cytokines promote interleukins-17A and 17F production by neutrophils during asthma[J]. Chinese Journal of Tissue Engineering Research, 2020, 24(31): 5044-5051.

Figures/Tables 8

2.6 白细胞介素21/白细胞介素23诱导的中性粒细胞中白细胞介素17产生及STAT3途径激活见图6。用白细胞介素21+23细胞因子刺激哮喘中性粒细胞 15 min，通过流式细胞术定量磷酸化激活的STAT3阳性细胞百分比(pSTAT3)。与未刺激的细胞相比(18.0%)，用白细胞介素21+白细胞介素23刺激后pSTAT3显著增加(44.3%，P=0.005)，见图6A。在存在Stattic(Sc-202818)的情况下刺激细胞后，pSTAT3的这种增加显著降低(20.1%，P=0.002)。用白细胞介素21+23或白细胞介素6细胞因子刺激健康的中性粒细胞可上调pSTAT3水平，但不显著。使用Sc-202818可抑制白细胞介素21+白细胞介素23诱导的磷酸化，见图6A，B。在存在或不存在Sc-202818的情况下，细胞被饥饿并用白细胞介素21+白细胞介素23细胞因子刺激，然后通过流式细胞术测定表达白细胞介素17A和白细胞介素17F的细胞的频率。在缺乏白细胞介素21+白细胞介素23细胞因子刺激神经营养素时，观察到白细胞介素17A阳性细胞显著增加(22.3% vs. 白细胞介素21+白细胞介素23：42.6%，P=0.011)抑制剂，见图6C。对于白细胞介素17F观察到类似的结果(27.50% vs. 白细胞介素21+白细胞介素23：55.3%，P=0.009)，见图6D。然而，当使用Sc-202818抑制STAT3磷酸化时，白细胞介素17A阳性细胞明显减少(22.3% vs. 白细胞介素21+白细胞介素23：23.5%，P=0.020)和白细胞介素17F(观察到27.5% vs. 白细胞介素21+白细胞介素23：18.6%，P=0.003)，表明白细胞介素21和白细胞介素23诱导的中性粒细胞中白细胞介素17的表达需要STAT3磷酸化，见图6C，D。直方图反映了在存在或不存在Sc-202818的情况下用白细胞介素21+白细胞介素23刺激后白细胞介素17A和白细胞介素17F表达的哮喘细胞百分比，见图6E，F。 "

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