新生期小鼠CTLA4Ig基因修饰的树突细胞对其成年期哮喘免疫耐受的影响  

Experimental study of CTLA4Ig gene-modified dendritic cells transfused neonatally inducing asthmatic immune tolerance in adult mice

作　　者：张瑾[1] 陈超[1] 陈艳[1] 王立波[1] 张晓波[1] 张明智[1] 

机构地区：[1]复旦大学附属儿科医院呼吸科,上海201102

出　　处：《中华医学杂志》2012年第12期848-852,共5页National Medical Journal of China

基　　金：国家自然科学基金（30901623）

摘　　要：目的探讨腺病毒重组CTLA4Ig基因修饰的树突细胞疫苗（AdCTLA4Ig-DC）在Th2细胞占主导地位的过敏性哮喘中的免疫调节机制。方法新生BALB／c雌性小鼠20只，抽签法随机分成AdCTLA4Ig．DC干预组和空载腺病毒转染DC（AdGFP—DC）干预组，各10只，分别回输AdCTLA4Ig-DC和AdGFP—DC，无特定病原体（SPF）环境下饲养至6～8周；另相同环境生长6—8周龄BALB／c雌性小鼠20只，抽签法随机分为空白对照组、哮喘组，各10只。空白对照组小鼠以生理盐水腹腔注射，并以雾化吸入，其余3组小鼠以卵白蛋白腹腔注射致敏，并以雾化吸入激发，通过观察各组小鼠激发时的症状表现、肺泡灌洗液（BALF）中嗜酸粒细胞比例和肺部病理切片改变了解小鼠气道炎症情况；酶联免疫吸附（ELISA）方法测定血清及BALF中IgE、白细胞介素（IL）-4、IL-10及干扰素（IFN）-1水平。结果哮喘组小鼠致敏后出现呼吸加快等哮喘表现、肺部病理表现为炎性浸润。AdCTLA4Ig．DC干预组小鼠成年期哮喘发作的症状明显减轻，且支气管黏膜下和周围肺组织的炎性细胞浸润减少；血清和BALF中IgE、IL_4水平均明显低于哮喘组[IgE：（3．9±1．3）×10^5比（5．34-1．7）×10^5pg／ml和（22．9±11．8）×10^3比（122．0±59．6）×10^3pg／ml，IL-4：（70．5±7．0）比（88．7±9．2）pg／ml和（46．9±2．9）比（73．3±8．7）pg／ml，均P〈0．05]，IL一10水平则均明显高于哮喘组[（442±8）比（227．4-11）pg／ml和（83±42）比（27±11）pg／ml，均P〈0．05]，IFN-γ水平与哮喘组差异无统计学意义（均P〉0．05）；BALF中嗜酸性细胞明显低于哮喘组[（18．5±1．9）×10^4／ml比（62．3±6．7）×10^4／ml，P〈0．05]。结论新生小鼠给予CTLA4Ig基因修饰的未成熟DC，能够抑制成年期卵白蛋白致敏、激发引起的Th2细胞活化，可诱导哮喘免疫耐受的发生。Objective To explore the effects of CTLA4Ig gene-modified dendritic cells on the mechanism of immune regulation in asthma with Th2 superiority. Methods Newborn BALB/e mice are randomly divided into 2 groups of AdCTLA4Ig-DC intervention and AdGFP modified dendritic cells （ AdGFP- DC） intervention by randomization （n = 10 each）. CTLA4Ig gene-modified dendritic cells （AdCTLA4Ig- DC） and AdGFP-DC were infused separately. Newborn BALB/c mice in both groups were fed for 6 - 8 weeks. Additional 6 - 8 weeks BALB/c mice were randomly divided into 2 groups of control and asthma （n = 10 each）. The saline-sensitized/challenged mice received an intraperitoneal injection while inhalation was administered in the control group. Ovalbumin （OVA） was used similarly in the remaining three groups. The manifestations of OVA challenged mice in each group, bronchoalveolar lavage fluid （BALF） eosinophils and pathological changes in airway were observed to examine the effects of CTLA4Ig gene-modified dendritic cells on airway inflammation. The levels of IgE, Interleukin （IL）-4, IL-10 and IFN-γ were detected by enzyme-linked immunosorbent assay （ELISA）. Results Rapid respiration and pulmonary inflammatory infiltration were obvious in the asthma group. CTLA4Ig gene-modified dendritic cells transfused neonatally significantly improved the symptoms of asthma in adulthood and reduced the infiltration of inflammatory cells in bronchial submueosa. The levels of IgE and IL-4 in serum and BALF in the AdCTLA4Ig-DC intervention group were significantly lower than those in the asthma group （ IgE ： （ 3.9 ± 1.3 ） x 105 pg/ml vs （ 5.3 ＋ 1.7） x 105 pg/ml and （22. 9 ＋ 11.8 ） x 103 pg/ml vs （ 122. 0 ± 59. 6） x 103 pg/ml, IL-4 ： （70. 5 ± 7.0） pg/ml vs （88. 7 ±9. 2） pg/ml and （46. 9 ±2. 9） pg/ml vs （73.3 ±8.7） pg/ml, all P 〈0. 05）. The levels of IL-10 in serum and BALF in the AdCTLA4Ig-DC intervention group were significantly higher than those in the asthma group （ （442

关 键 词：小鼠 哮喘 树突细胞 免疫耐受 模型 动物 

分 类 号：R562.25[医药卫生—呼吸系统]