抗CD20单克隆抗体对哮喘小鼠气道炎症的影响
Effect of anti-CD20 monoclonal antibody on airway inflammation in a murine model of asthma
贺基龙 1章宗悦 1左小淑 1林琪斌 1倪海阳 1丁续红 1黄毅 1余红缨 1聂汉祥1
作者信息
- 1. 武汉大学人民医院呼吸与危重症医学科 湖北 武汉 430060
- 折叠
摘要
目的:观察抗CD20单克隆抗体(简称抗CD20单抗)对哮喘小鼠气道炎症的影响.方法:将15只6～8周龄的BALB/c雌性小鼠随机分成正常对照组、哮喘对照组和抗CD20单抗治疗组,每组 5只.哮喘对照组和抗CD20单抗治疗组以屋尘螨(HDM)致敏和激发,正常对照组以等量PBS代替,其中抗CD20单抗治疗组在第1次激发前2 d给予抗CD20单抗经鼻滴入联合腹腔注射,正常对照组和哮喘对照组给予等量同型对照抗体代替.苏木精-伊红(HE)染色和糖原(PAS)染色观察肺部炎症和气道杯状细胞增生;瑞氏-吉姆萨染色检测支气管肺泡灌洗液(BALF)细胞总数和分类计数;酶联免疫吸附试验检测血清HDM特异性IgE及BALF中IL-4、IL-5、IL-13、IFN-γ和IL-10水平;流式细胞术检测肺CD19+B细胞数量.结果:与哮喘对照组比较,抗CD20单抗治疗组哮喘小鼠肺组织炎性细胞浸润减少,气道基底膜杯状细胞数量以及BALF中细胞总数、嗜酸粒细胞、巨噬细胞和淋巴细胞数量明显减少(P<0.01);血清HDM特异性IgE及BALF中IL-4、IL-5和IL-13水平明显降低(P<0.01);BALF中IFN-γ水平无统计学意义(P>0.05);BALF中IL-10水平明显增高(P<0.01).同时,抗CD20单抗治疗组哮喘小鼠肺CD19+B细胞数量明显减少(P<0.01).结论:经鼻滴入联合腹腔注射抗CD20单抗可以减轻哮喘小鼠气道炎症,可能与其去除肺B细胞导致气道IL-10水平增高有关.
Abstract
objective:To investigate the effect of anti-CD20 monoclonal antibody(mAb)on airway inflam-mation in a murine model of asthma.Methods:Fifteen female BALB/c mice aged 6-8 weeks were randomly divided into the normal control group,asthma control group,and anti-CD20 mAb treatment group,with 5 mice in each group.The asthma group and the anti-CD20 mAb treatment group were sensitized and challenged with house dust mite(HDM),while the normal control group was replaced with an equivalent dose of PBS.The anti-CD20 mAb was given by intranasal drip combined with in-traperitoneal injection two days before the first challenge in the anti-CD20 mAb treatment group,and an equal amount of IgG2c isotype control was given instead in the normal control group and asthma group.Hematoxylin-eosin(HE)staining and periodic acid-schiff(PAS)staining were used to assess lung histopathology and airway goblet cell hyperplasia.Total and classified counts of cells in bronchial alveolar lavage fluid(BALF)analyzed by Wright-Giemsa staining.Enzyme-linked immunosorbent as-say(ELISA)was adopted for the detection of serum HDM-specific IgE,IL-4,IL-5,IL-13,IFN-γ,and IL-10 levels in BALF.The number of CD19+B cells in the lungs was detected by flow cytome-try.Results:Compared with those in the asthma group,lung tissue inflammatory cell infiltration and the number of airway goblet cells in asthmatic mice in the anti-CD20 mAb treatment group were significantly reduced,as well as the total number of cells,eosinophils,macrophages,and lymphocytes in BALF(P<0.01);serum HDM-specific IgE and IL-4,IL-5 and IL-13 levels in BALF were signifi-cantly reduced(P<0.01);no statistically significant changes was found in IFN-γ levels in BALF(P>0.05);and IL-10 levels in BALF were significantly increased(P<0.01).Meanwhile,the number of lung CD19+B cells in asthmatic mice in the anti-CD20 mAb treatment group was significantly reduced(P<0.01).Conclusion:The combination of intranasal drip and intraperitoneal injection of anti-CD20 mAb attenuates airway inflammation in a murine model of asthma,which might be associated with in-creased airway IL-10 level.
关键词
哮喘/抗CD20单克隆抗体/气道炎症/B细胞/白细胞介素-10Key words
Asthma/Anti-CD20 Monoclonal Antibody/Airway Inflammation/B Cells/Inter-leukin-10引用本文复制引用
基金项目
国家自然科学基金资助项目(81970024)
出版年
2024
NETL
NSTL
万方数据