热带螨重组Blo t 5变应原诱导小鼠过敏模型的建立

doi:10.16431/j.cnki.1671-7236.2017.07.038

《中国畜牧兽医》 ›› 2017, Vol. 44 ›› Issue (7): 2171-2177.doi: 10.16431/j.cnki.1671-7236.2017.07.038

热带螨重组Blo t 5变应原诱导小鼠过敏模型的建立

李晓烨¹, 肖正泮¹, 刘楚¹, 韦双双², 王大勇², 裴业春¹

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

修回日期:2017-04-10 出版日期:2017-07-20 发布日期:2017-07-22
通讯作者: 裴业春 E-mail:ycpei@hainu.edu.cn
作者简介:李晓烨(1994-),女,浙江绍兴人,本科,研究方向:生物技术与分子药理学,E-mail:756997565@qq.com
基金资助:
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）

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

LI Xiao-ye¹, XIAO Zheng-pan¹, LIU Chu¹, WEI Shuang-shuang², WANG Da-yong², PEI Ye-chun¹

1. College of Animal Science and Technology, Institute of Tropical Agriculture and Forestry, Hainan University, Haikou 570228, China;
2. College of Biological Science, Institute of Tropical Agriculture and Forestry, Hainan University, Haikou 570228, China

Revised:2017-04-10 Online:2017-07-20 Published:2017-07-22

摘要/Abstract

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。

关键词: 热带螨; 重组Blo t 5蛋白; 过敏模型

Abstract:

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 PQE80L. 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 5 group, 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 5 group (P<0.05).The mice in rBlo t 5 group showed inflammatory cell infiltration in airway by HE.These results showed that the allergic murine model was induced successfully by rBlo t 5 allergen.

Key words: Blomia tropicalis; recombinant Blo t 5 (rBlo t 5); allergic model

中图分类号:

R392.8

李晓烨, 肖正泮, 刘楚, 韦双双, 王大勇, 裴业春. 热带螨重组Blo t 5变应原诱导小鼠过敏模型的建立[J]. 《中国畜牧兽医》, 2017, 44(7): 2171-2177.

LI Xiao-ye, XIAO Zheng-pan, LIU Chu, WEI Shuang-shuang, WANG Da-yong, PEI Ye-chun. Development of an Allergic Murine Model Induced by Recombinant Blo t 5 Allergen of Blomia tropicalis[J]. , 2017, 44(7): 2171-2177.

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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

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