摘要
目的观察以聚乳酸-羟基乙酸共聚物(PLGA)材料为佐剂制备的重组屋尘螨2类变应原(rDerp2)纳米微粒疫苗(DEPN)对小鼠过敏性气道炎症的影响,并探讨其免疫治疗机制。方法制备PLGA-rDerp2纳米粒子并鉴定其特性。40只BALB/c小鼠随机分为5组,A组(对照组)均给予生理盐水(100μl)。B、C、D和E组腹部皮下注射屋尘螨粗浸液(10μg)免疫小鼠致敏,然后分别用PBS(100μl)、2mg空白PLGA粒子(emptyPLGA,EP)、100μgrDerp2、2mg的DEPN纳米疫苗(载有100μgrDerp2)皮下注射进行免疫治疗,连续免疫治疗3d,1次/d,各组用rDerp2(50μg)滴鼻激发,激发后第2天剖杀,收集支气管肺泡灌洗液(BALF)并对细胞进行总计数和分类计数;HE染色和PAS染色(PeriodicAcid-SchiffStain)观察小鼠肺部组织炎症和支气管黏液分泌;用ELISA检测BALF和脾细胞培养上清的细胞因子(IL-4、IFN-γ)和血清中变应原特异性IgG2a和IgE抗体浓度。结果B、C组肺部呈明显的变态反应性炎症,D、E组变应原诱导的肺部嗜酸粒细胞浸润和黏液分泌比B、C组显著减轻。BALF中的细胞总数B组比A组明显增多,分类细胞以中性和嗜酸粒细胞为主,超过50%。rDerp2特异性IgE抗体水平,D组(0.93±0.04)和E组(0.77±0.10)均低于B组(1.14±0.10)(P<0.01);特异性IgG2a抗体水平,D组(1.02±0.01)和E组(1.17±0.46)均高于B组(0.14±0.01)(P<0.01)。在BALF中,D组[(55.60±3.79)pg/ml]和E组[(48.60±4.50)pg/ml]IL-4水平均低于B组[(78.90±6.07)pg/ml](P<0.01);IFN-γ水平E组[(68.50±2.87)pg/ml]显著高于B组[(27.30±3.51)pg/ml](P<0.01)。脾细胞上清的IL-4水平,D组[(56.3±4.85)pg/ml]和E组[(40.2±4.36)pg/ml]显著低于B组[(81.20±6.84)pg/ml](P<0.01);IFN-γ水平,E组[(70.20±3.85)pg/ml]显著高于B组[(34.60±2.25)pg/ml]。结论DEPN免疫治疗可抑制小鼠肺部过敏炎症,其机制可能与调节Th1/Th2平衡有关。
Objective To investigate the efficacy and mechanism of subcutaneously given recombinant Der p 2 entrapped PLGA nanopartieles （DEPN） on mouse model with allergie airway inflammation. Methods 40 BALB/c mice were randomly divided into 5 groups, group A （normal eontrol） were treated with saline （100 μl） all the time, groups B, C, D and E were sensitized intraperitoneally with erude dust mite extracts （10 μg） and then subcutaneously treated respectively with PBS （100μl）, 2mg empty PLGA （EP）, 100μg rDerp2, and 2mg DEPN （loaded with 100 μg rDerp 2） for 3 times, once per day, followed by intranasal challenge of 50 μg rDer p 2. One day pest ehallenge, miee were saerifieed and bronehoalveolar lavage fluid （BALF） was eolleeted. Number of the total eells and eosinophils was determined, and airway inflammation and mueus seeretion were analyzed by haematoxylin and eosin （H＆E） staining and periodic aeid-Schiff （PAS） staining. Level of eytokines in the supernatant of sphnocyte culture was assayed by ELISA. Level of rDer p 2 specific IgG2a and IgE in the sera was determined by ELISA. Results The lung histology showed development of eosinophil infiltration in the airway of mice in groups B and C. The lung inflammation and mucus secretion in groups D and E were significantly alleviated than that of groups B and C. Number of total cells （63.50±5.12） and eosinophils （15.32±3.04） in BALF decreased in group B. Compared with group B, the number of total cells in groups D （55.3±5.20）×10^4 /ml and E （41.00±4.91）×10^4 /ml greatly decreased （P〈0.05）, and same with that of eosinophils in groups D （9.56±1.09）×10^4 /ml and E （3.22_±0.31）×10^4 /ml. The rDerp2 specific IgE and IgG2a antibodies in group B were 1.14±0.105 and 0.14±0.07 respectively. The level of specific IgE was significantly lower （P〈0.01） in groups D （0.93± 0.04） and E （0.77-±0.09）, and that of IgG2a in groups D （1.02±0.01） and E （1.17±0.46） were significantly high
出处
《中国寄生虫学与寄生虫病杂志》
CAS
CSCD
北大核心
2006年第6期414-419,共6页
Chinese Journal of Parasitology and Parasitic Diseases
基金
国家863高技术发展计划项目(No.2002AA214011)
国家自然科学基金(No.30471505
30271226)
粤港关键领域重点突破项目(No.20054982207)
广东省科技计划重大项目(No.2003C104019)
深圳市科技计划(No.200218)~~
