热带螨重组Blo t 5变应原诱导小鼠过敏模型的建立  

Development of an Allergic Murine Model Induced by Recombinant Blo t 5 Allergen of Blomia tropicalis

作　　者：李晓烨[1] 肖正泮 刘楚[1] 韦双双[2] 王大勇[2] 裴业春[1] 

机构地区：[1]海南大学热带农林学院动物科技学院,海口570228 [2]海南大学热带农林学院生物学院,海口570228

出　　处：《中国畜牧兽医》2017年第7期2171-2177,共7页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金青年项目(31402257);海南省科技项目(ZDYF2017110);海南省自然科学基金项目(20153084)

摘　　要：为了建立热带螨重组Blo t 5(rBlo t 5)变应原诱导小鼠过敏模型,试验对热带螨Blo t 5基因序列进行优化与全基因合成,构建了原核表达载体PQE80L-Blo t 5,并在大肠杆菌中进行表达。利用镍柱亲和层析法纯化了rBlo t 5蛋白,Western blotting结果显示,纯化的rBlo t 5蛋白能与Blo t 5鼠单克隆抗体结合。随后将12只BALB/c小鼠随机均分为对照组(PBS组)与诱导组(rBlo t 5组),通过腹腔注射致敏与气道诱导后,酶联免疫吸附试验(ELISA)检测血清总IgE水平;鼠尾静脉注射变应原诱发应激性过敏反应;HE染色观察小鼠肺部炎症严重程度。诱导小鼠过敏后发现,与PBS组相比,rBlo t 5组小鼠血清总IgE水平显著升高(P<0.05),应激性过敏反应引起体温显著下降(P<0.05),肺部组织细胞浸润显著增加(P<0.05)。本研究结果表明,原核表达纯化的rBlo t 5蛋白能够成功建立小鼠过敏模型。To development an allergic murine model induced by the recombinant Blo t 5（rBlo t 5）allergen,the Blo t 5 gene sequences were optimized,synthesized and inserted into the prokaryotic expression vector PQE80 L.The Blo t 5 was expressed in E.coli,purified by NI-NTA agarose,and tested by Western blotting.The results showed that the rBlo t 5 could specially react with monoclonal antibodies against Blo t 5.Twelve BALB/c mice were randomly divided into control group（PBS group,n=6）and rBlo t 5-induced group（rBlo t 5group,n=6）.After sensitization by intraperitoneal injection（i.p）and challenged by intratracheal instillation of rBlo t 5,the total serum IgE levels were measured by ELISA;The mice in rBlo t 5 group were challenged with rBlo t 5 by intraperitoneal injection,and the body temperature was measured at regular intervals;The lung was fixed and stained with haematoxylin and eosin（HE）.The result showed that compare with PBS group,the total serum IgE levels rised significantly in rBlo t 5 group（P0.05）.The body temperature dropped significantly and a severe allergic reaction（anaphylaxis）was occurred in mice of rBlo t 5group（P0.05）.The mice in rBlo t 5group showed inflammatory cell infiltration in airway by HE.These results showed that the allergic murine model was induced successfully by rBlo t 5 allergen.

关 键 词：热带螨 重组Blo t 5蛋白 过敏模型 

分 类 号：R392.8[医药卫生—免疫学]