摘要
目的建立重组德国小蠊变应原Blag2(rBlag2)诱导的小鼠变态反应气道炎症动物模型。方法在大肠杆菌中诱导表达Blag2重组蛋白,用镍亲和柱层析法提纯重组蛋白;24只BALB/c小鼠随机分为德国小蠊粗浸液组(A组)、德国小蠊粗浸液+重组Blag2组(B组)、重组Blag2(C组)、阴性对照组(D组)。分别通过HE染色和PAS染色(periodicacid-Schiff)观察小鼠肺部炎症和黏液分泌;观察支气管肺泡灌洗液(BALF)中细胞总数和细胞分类;用酶联免疫吸附试验(ELISA)检测BALF、脾细胞培养上清的细胞因子和血清rBlag2特异的IgE、IgG1、IgG2a抗体。结果成功表达、纯化Blag2重组蛋白;A、B与C组肺部病理改变呈现明显的变态反应性炎症;BALF中的细胞总数、中性粒细胞数、EOS计数、IL-4、血清抗原特异性IgE抗体、IgG1抗体和脾细胞分泌IL-4均显著高于阴性对照组(P<0.01)。结论用rBlag2能成功建立小鼠变态反应气道炎症动物模型。
Objective To develop a murine model of allergic airway inflammation by recombinant Blatella germanica allergen-rBla g2 as clinical relevant allergen. Methods The recombinant Bla g2 was expressed in E. coli bacteria and purified through Ni^＋ NTA His agarose. Twenty-four BALB/c mice were divided randomly into crude extract of German cockroach group（group A, n = 6）, rBla g2 ＋ crude extract of German cockroach group （group B, n = 6）, rBla g2 group（group C, n = 6） and control group（group D, n = 6）. After sensitization by intraperitoneal injection （ i. p） and challenged by intranasal instillation of rBla g2, the lung was fixed and stained with haematoxylin and eosin（H＆E）. The lung tissues were also stained with periodic acid-Schiff（PAS） to evaluate the degree of proliferation of goblet cells. The total cell number and composition of BALF samples were determined. The cytokines in splenocytes culture supernatants and BALF were assayed by enzyme-linked immunoassay （ELISA）. rBla g2 specific IgG1, IgG2a and IgE in serum were also measured by ELISA. Results Mice in model A, B and C group showed inflammatory cell infiltration in airway. Total cells, neutrophils, eosinophils, and IL-4 in BALF; allergen-specific antibody IgE and IgG1 in sertum; IL-4 released from splenocytes into supemtants in vitro were all significantly increased in group A, B, C as compared with those in group D（P〈 0.01）. This model may be exploited in the study of pathophysiological and immunopathlogical mechanisms.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2006年第5期446-451,共6页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金(No.39860071
No.30571625)
广东省科技计划重点项目(No.2003A3080502)
深圳市科技计划项目
