中山大学附属第一医院耳鼻咽喉科医院,广东 广州 510080
方淑斌,博士后,研究方向:呼吸道变应炎症及感音神经性聋的发病机制及其干预,E-mail:fangshb3@mail.sysu.edu.cn
收稿:2021-05-25,
纸质出版:2021-11-20
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方淑斌,付清玲.变应性鼻炎患者血浆细胞外囊泡对树突状细胞和巨噬细胞的调控作用[J].中山大学学报(医学科学版),2021,42(06):808-815.
FANG Shu-bin,FU Qing-ling.Effects of Plasma Extracellular Vesicles on Dendritic Cells and Macrophages in Patients with Allergic Rhinitis[J].Journal of Sun Yat-sen University(Medical Sciences),2021,42(06):808-815.
方淑斌,付清玲.变应性鼻炎患者血浆细胞外囊泡对树突状细胞和巨噬细胞的调控作用[J].中山大学学报(医学科学版),2021,42(06):808-815. DOI: 10.13471/j.cnki.j.sun.yat-sen.univ(med.sci).2021.0602.
FANG Shu-bin,FU Qing-ling.Effects of Plasma Extracellular Vesicles on Dendritic Cells and Macrophages in Patients with Allergic Rhinitis[J].Journal of Sun Yat-sen University(Medical Sciences),2021,42(06):808-815. DOI: 10.13471/j.cnki.j.sun.yat-sen.univ(med.sci).2021.0602.
目的
2
树突状细胞(DCs)和巨噬细胞(Mφ)在变应性鼻炎(allergic rhinitis, AR)的发病过程中起重要调控作用。本研究主要探讨AR血浆细胞外囊泡(EVs)对DCs和Mφ分化的调控作用。
方法
2
招募健康志愿者(HC)和AR患者各6人,应用超速离心法制备HC-EVs和AR-EVs;将磁珠分选所得人CD14
+
单核细胞在体外分别诱导分化成DCs和Mφ,并用5×10
8
/mL HC-EVs和AR-EVs分别对单核细胞、DCs和Mφ进行刺激,通过ELISA、聚合酶链反应(PCR)或流式分析的方法进行分析。
结果
2
流式分析表明,与HC-EVs相比,AR-EVs刺激CD14
+
单核细胞5天后,单核细胞表面CD14、CD68、CD80和CD86表达水平均无显著差异。用AR-EVs刺激不成熟DCs(iDC)2天后,iDC表面HLA-DR、CD40和CD80表达水平亦无显著差异。用AR-EVs刺激Mφ 2天后,PCR显示Mφ中IL-6(
P
<0.001)和TNF-α(
P
<0.05)的表达水平显著下降,TGM2的表达水平显著上升(
P
<0.05),而MRC1和IL-10的表达水平则无显著变化。
结论
2
AR-EVs可显著促进Mφ向促炎的M2型极化,抑制Mφ向M1型极化,但AR-EVs对CD14
+
单核细胞向DCs和Mφ分化及DCs的成熟均无明显作用,这提示血浆EVs可能通过调控Mφ的极化促进AR的发生发展。
Objectives
2
Dendritic cells (DCs) and macrophages (DCs) are involved in the pathogenesis of allergic rhinitis (AR). The present study was aimed to explore the effects of plasma extracellular vesicles (EVs) derived from patients with AR on differentiation of DCs and Mφ.
Methods
2
Healthy control (
n
=6, HC) and AR patients (
n
=6) were recruited, and HC-EVs and AR-EVs were isolated from plasma by ultracentrifugation. Human CD14
+
monocytes were isolated and induced into DCs and Mφ, respectively. Human monocytes, DCs and Mφ were cultured in the presence of 5×10
8
/mL HC-EVs or AR-EVs, and collected for polymerase chain reaction (PCR) or flow cytometry analysis.
Results
2
Compared with HC-EVs, no significant effect was observed on levels of CD14, CD68, CD80 and CD86 on monocytes after being treated with AR-EVs for 5 days. Similarly, levels of HLA-DR, CD40 and CD80 on immature DCs (iDCs) showed no significant difference in the presence of AR-EVs for 2 days. Interestingly, levels of IL-6 (
P
<0.001) and TNF-α (
P
<0.05) were significantly decreased, and level of TGM2 were significantly increased in Mφ treated with AR-EVs for 2 day. However, no effects of AR-EVs were observed on levels of MRC1 and IL-10.
Conclusion
2
AR-EVs were able to promote proinflammatory M2 polarization and inhibit M1 polarization of Mφ, but showed no significant effects on differentiation of DCs or Mφ as well as the maturation of DCs, suggesting that plasma EVs could promote the development of AR via regulation on Mφ polarization.
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