作 者：
李晓烨;肖正泮;刘楚;韦双双;王大勇;裴业春
单 位：
海南大学热带农林学院动物科技学院;海南大学热带农林学院生物学院
关键词：
热带螨;重组Blo t 5蛋白;过敏模型
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。
译 名：
Development of an Allergic Murine Model Induced by Recombinant Blo t 5 Allergen of Blomia tropicalis
作 者：
LI Xiao-ye;XIAO Zheng-pan;LIU Chu;WEI Shuang-shuang;WANG Da-yong;PEI Ye-chun;College of Animal Science and Technology,Institute of Tropical Agriculture and Forestry,Hainan University;College of Biological Science,Institute of Tropical Agriculture and Forestry,Hainan University;
关键词：
Blomia tropicalis;;recombinant Blo t 5(rBlo t 5);;allergic model
摘 要：
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(P<0.05).The body temperature dropped significantly and a severe allergic reaction(anaphylaxis)was occurred in mice of rBlo t 5group(P<0.05).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.
