热带螨重组Blo t 5变应原诱导小鼠过敏模型的建立

发布时间：2018-06-25 10:17

  本文选题：热带螨 + 重组Blo　； 参考：《中国畜牧兽医》2017年07期

【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(P0.05),应激性过敏反应引起体温显著下降(P0.05),肺部组织细胞浸润显著增加(P0.05)。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。
[Abstract]:In order to establish a mouse allergic model induced by recombinant Blo t 5 (rBlo t 5), a prokaryotic expression vector PQE80L-Blo t 5 was constructed and expressed in Escherichia coli. The purified rBlo t 5 protein was purified by nickel column affinity chromatography. The results showed that the purified rBlo t 5 protein could bind to the monoclonal antibody of Blo t 5 mouse. Then 12 BALB / c mice were randomly divided into two groups: control group (PBS group) and induction group (rBlo t 5 group). The serum IgE levels were measured by enzyme-linked immunosorbent assay (Elisa) after intraperitoneal injection of sensitization and airway induction. The severity of lung inflammation in mice was observed by HE staining. Compared with PBS group, the serum total IgE level of mice induced by allergy was significantly higher than that of PBS group (P0.05), the body temperature of stress allergic reaction decreased significantly (P0.05), and the infiltration of lung tissue cells increased significantly (P0.05). The results showed that the purified rBlo t 5 protein could successfully establish mouse allergic model.
【作者单位】： 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;
【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)
【分类号】：R-332;R392
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本文编号：2065578