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中华妇幼临床医学杂志(电子版) ›› 2011, Vol. 07 ›› Issue (06) : 546 -551. doi: 10.3877/cma.j.issn.1673-5250.2011.06.008

论著

肽聚糖对变态反应性哮喘产妇新生儿脐带血中嗜碱性粒细胞的作用
杨玲, 许以平, 郑晨, 刘鸣, 熊瑛, 祝捷, 史桂英, 陈玉英   
  1. 200092 上海,上海交通大学医学院附属新华医院
    上海交通大学医学院附属仁济医院
    上海交通大学医学院细胞生物学教研室
  • 出版日期:2011-12-01

Effect of Peptidoglycan on Umbilical Cord Blood-Derived Basophils From Maternal With Allergic Asthma

Ling YANG, Yi-ping XU, Chen ZHENG, Ming LIU, Ying XIONG, Jie ZHU, Gui-ying SHI, Yu-ying CHEN   

  1. Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine, 200092 Shanghai, China
  • Published:2011-12-01
  • Supported by:
    * Project No. YG2010MS12, supported by the Biomedical Engineering Cooperation Foundation of Shanghai Jiaotong University
引用本文:

杨玲, 许以平, 郑晨, 刘鸣, 熊瑛, 祝捷, 史桂英, 陈玉英. 肽聚糖对变态反应性哮喘产妇新生儿脐带血中嗜碱性粒细胞的作用[J]. 中华妇幼临床医学杂志(电子版), 2011, 07(06): 546-551.

Ling YANG, Yi-ping XU, Chen ZHENG, Ming LIU, Ying XIONG, Jie ZHU, Gui-ying SHI, Yu-ying CHEN. Effect of Peptidoglycan on Umbilical Cord Blood-Derived Basophils From Maternal With Allergic Asthma[J]. Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition), 2011, 07(06): 546-551.

目的

通过采用肽聚糖(PGN)刺激变态反应性哮喘产妇新生儿脐带血中嗜碱性粒细胞,测定其Toll样受体2(TLR2)的表达及白细胞介素(IL)-4的分泌,探讨PGN对变态反应性哮喘产妇新生儿脐带血中嗜碱性粒细胞的作用。

方法

自2008年3月1日至2009年3月1日在上海交通大学医学院附属新华医院住院分娩的12例变态反应性哮喘产妇(哮喘组)和16例健康产妇(对照组)的新生儿脐带血中提纯嗜碱性粒细胞,采用PGN刺激,经TLR2荧光抗体标记后,通过流式细胞术从抗体水平测定脐带血中嗜碱性粒细胞的TLR2表达;采用原位杂交技术从核酸水平观察TLR2 mRNA表达;采用ELISA法检测培养的上清液中IL-4含量(本研究遵循的程序符合本院人体试验委员会所制定的伦理学标准,得到该委员会批准,分组征得受试对象本人的知情同意,并与之签署临床研究知情同意书)。两组孕妇年龄、分娩方式、分娩孕龄及脐带血采集方式等比较,差异无统计学意义(P>0.05)。

结果

流式细胞术检测及原位杂交结果显示,采取PGN刺激后,哮喘组新生儿脐带血中嗜碱性粒细胞TLR2蛋白表达及核酸表达均显著高于对照组,两组比较,差异有统计学意义(P<0.01);上清液中IL-4的含量明显高于对照组,差异亦有统计学意义(P<0.01)。

结论

TLR2介导的天然免疫反应通过嗜碱性粒细胞释放介质而导致变态反应性炎症发生。细菌成分可通过TLR2受体激活嗜碱性粒细胞(尤其有变态反应性疾病遗传史时)释放介质,可能是变态反应性疾病发病增加的原因。

Objective

To determine if basophils harvested from neonates genetically predisposed to atopic disease had different levels of Toll-like receptors (TLR)2 expression and investigate whether putative TLR2 ligands mediated cytokine secretion.

Methods

Blood samples were collected from 12 asthmatic and 16 healthy women and their newborns. Informed consent was obtained from all participates. TLR2 expression was determined by using nucleic acid hybridization in situ and flow cytometry. IL-4 levels were quantified by ELISA following peptidoglycan (PGN) stimulation.

Results

TLR2 expression at the protein level and mRNA level were up-regulated in cord blood basophils from neonates born to mothers with allergic asthma following stimulation with PGN. Basophils purified from the cord blood of neonates born to atopic mothers produced more IL-4 compared to basophils purified from children born to non-atopic controls(P<0.01).

Conclusion

These data TLR2-mediated innate immune responses play a role in augmenting allergic reactions via modulation of basophil cytokine secretion. Microbial components may activate basophils through TLR2 (especially for genetically predisposed infants) to release cytokines associated with an increased incidence of allergic diseases.

图1 流式细胞仪分选嗜碱粒细胞(A:选取SSC低信号细胞;B:CD203c-PE+ CD45-PerCP+的细胞P5区即为嗜碱粒细胞)
图2 嗜碱粒细胞阿利新蓝染色(箭头所指为嗜碱粒细胞)(×400)
图3 哮喘组嗜碱粒细胞经PGN刺激前、后TLR2表达(散点图)
图4 PGN刺激前,哮喘组新生儿脐带血嗜碱粒细胞TLR2mRNA表 达(绿色箭头所指,呈浅蓝色)
图5 PGN刺激后,哮喘组新生儿脐带血嗜碱粒细胞TLR2mRNA表 达(红色箭头所指,呈浅棕色)
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