屋尘螨致敏小鼠鼻炎模型建立及其免疫学评价  

Establishment of house dust mite(Dermatophagoides pteronyssinus)induced allergic rhinitis mouse model and research on related immunological evaluation

作　　者：张影[1] 徐颖华[1,2] 江霞云 鲁旭 杨蕾[1] 於洋[1] 王斌 赵爱华[1] 李丽莉[3] ZHANG Ying;XU Ying-hua;JIANG Xia-yun;LU Xu;YANG Lei;YU Yang;WANG Bing;ZHAO Ai-hua;LI Li-li(NHC Key Laboratory for Research on Quality and Standardization of Biotech Products,National Institute for Food and Drug Control,Beijing 102629,China;National Center for Medical Culture Collections,Beijing 102629,China;Institute for in Vitro Diagnostic Control,National Institute for Food and Drug Control,Beijing 100050,China)

机构地区：[1]中国食品药品检定研究院,国家卫生健康委员会生物技术产品检定方法及其标准化重点实验室,北京102629 [2]中国医学细菌保藏管理中心,北京102629 [3]中国食品药品检定研究院体外诊断试剂检定所,北京100050

出　　处：《中华临床免疫和变态反应杂志》2021年第6期608-617,共10页Chinese Journal of Allergy & Clinical Immunology

基　　金：学科带头人培养基金[2019X5]。

摘　　要：目的基于屋尘螨变应原组主要变应原(Der.p1)的剂量建立稳定的屋尘螨过敏小鼠模型,并以此系统筛选过敏评价生物标识。方法以屋尘螨变应原提取物为致敏蛋白,以氢氧化铝为佐剂,依据主要变应原(Der.p1)含量设置不同抗原浓度组和不同免疫针次,皮下注射免疫BALB/c小鼠,末次免疫一周后,采用Der.p1浓度为500μg/mL的屋尘螨变应原提取物滴鼻激发,每只小鼠20μL,每天1次,持续1周。通过观察小鼠的过敏反应症状、检测鼻部组织病理变化、测定各组小鼠体液免疫和细胞免疫指标的变化,筛选最佳的过敏模型建立程序。在此基础上更换不同来源屋尘螨抗原重新致敏BALB/c小鼠,验证过敏小鼠模型建立方法的可重复性。结果致敏小鼠经抗原激发后均出现明显的过敏反应,其中最佳免疫程序为Der.p1浓度100μg/mL、皮下注射两次。该致敏组小鼠经抗原激发后体内IL-4特异性淋巴细胞数(174±23)明显增多,与阴性对照组(22±0.5)比较差异具有统计学意义(P=0.011);脾淋巴细胞Th2型极化显著(Th1/Th2比值为0.362±0.028),与阴性对照组(0.832±0.07)比较差异具有统计学意义(P=0.034);血清屋尘螨特异性IgE抗体水平与激发前比较明显增高(P=0.00017),总IgE抗体水平与阴性对照组比较明显增高(P=0.041)。利用不同来源的屋尘螨抗原,选择优化的致敏程序建立过敏小鼠模型,表现出较好的可重现性。结论成功构建屋尘螨变应原特异性致敏小鼠模型,并以此筛选到致敏相关的生物标识,证实Der.p1剂量与致敏效果间的量效关系,为后续屋尘螨过敏症机制研究及屋尘螨变应原脱敏制剂研制提供实验基础。Objective To established a stable house dust mite(Dermatophagoides pteronyssinus)induced allergic mouse model basing on dose of Der p1,major allergen of Dermatophagoides pteronyssinus,for screening biomarkers systematically.Methods Dermatophagoides pteronyssinus induced allergic mouse model was established,based on different subcutaneous immunization protocols by using BALB/C mouse,in which Dermatophagoides pteronyssinus extract was used as sensitizing protein and aluminum hydroxide as adjuvant.After last injection of immunization,Dermatophagoides pteronyssinus extract contained 500μg/mL Derp1 was used for nasal provocation,20μL per time per mice,once a day for one week.The best protocol for establishment of Dermatophagoides pteronyssinus induced allergic mouse model was selected,basing on allergic reaction symptoms,pathological changes of nasal tissues and alternations of humoral and cellular immunological parameters of mice in each group.Reproducibility of the selected protocol for establishment of allergic rhinitis mouse model was verified,by using different house dust mite extracts for sensitization.Results Obvious allergic reaction was observed among sensitized mice after allergen provocation,while IL-4 secreting lymphocytes increased significantly,Th2 polarization of spleen lymphocyte was remarkable and serum Dermatophagoides pteronyssinus specific IgE and total IgE levels increased significantly among them.Allergic rhinitis mouse model was established by using Dermatophagoides pteronyssinus extracts from different sources and good reproducibility was demonstrated.Conclusions House dust mite(Dermatophagoides pteronyssinus)induced allergic rhinitis mouse model was established and related biomarkers for sensitization were screened successfully,as well as dose-effect relationship between Der.p1 dose and sensitization effect was demonstrated,providing experimental basis for further research on mechanism of house dust mite allergy and development of house dust mite extract products for immunotherapy.

关 键 词：屋尘螨 Der.p1 过敏 动物模型 免疫反应 

分 类 号：R28[医药卫生—中药学]