大鼠嗜碱性白血病粒细胞系RBL-2H3和RBL-1细胞模型在评价食品潜在致敏性中的比较  

Comparison of Rat(Rattus norvegicus) Basophilic Leukaemia(RBL)-2H3 and RBL-1 Cell Models for Evaluating Potential Allergenicity of Foods

作　　者：孙娜[1] 周催[1] 王翠燕[1] 孙璐[1] 车会莲[1] 

机构地区：[1]中国农业大学食品科学与营养工程学院,北京100083

出　　处：《农业生物技术学报》2013年第12期1465-1472,共8页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金面上项目(No.81273077)

摘　　要：目前广泛通过检测过敏患者血清中过敏原特异性IgE含量来进行过敏诊断，然而有研究显示，过敏患者血清中IgE水平与临床症状并无直接的关系。本研究通过比较大鼠（Rattus norvegicus） 嗜碱性白血病粒细胞系（rat basophilleukemia, RBL） RBL-2H3和RBL-1细胞与致敏小鼠（Mus musculus）血清中免疫球蛋白（IgE）抗体的结合能力以及脱颗粒能力，建立一种用于评价食品蛋白潜在致敏性的细胞免疫学评价方法。采用3种具有不同潜在致敏性的食品蛋白（大豆球蛋白（Gly）、卵清白蛋白（OVA）和马铃薯酸性磷酸酶（PAP）），对3~4周龄雌性Balb/c小鼠进行经口致敏5次，获取致敏血清用于流式细胞术对RBL-2H3和RBL-1细胞结合能力的分析以及体外介质释放实验。同时测定Balb/c小鼠的系统性过敏反应，探究RBL细胞脱颗粒反应与系统性过敏反应的关系。结果显示，小鼠血清IgE与RBL-2H3细胞的结合能力显著高于RBL-1细胞（P＜0.05）。RBL-2H3细胞表现出较好的脱颗粒反应，显著高于RBL-1细胞（P＜0.05），而RBL-1细胞表现出较低的脱颗粒反应。另外，RBL-2H3细胞介质释放实验检测灵敏度高，只需10~100 ng/mL的过敏原即可诱发细胞产生最强的脱颗粒反应，能够检测到食品中微量的致敏蛋白，检测特异性高；用抗OVA的小鼠血清致敏细胞，Gly诱导后，细胞未发生脱颗粒，能够区分Gly、OVA和PAP 3种不同致敏性的食品蛋白，与系统性过敏反应结果一致，真正反映食品蛋白诱发IgE介导的过敏反应的能力。研究结果提示，RBL-2H3细胞模型可以作为一种用于检测食品蛋白潜在致敏活性的有效方法。研究结果为建立评价新型食物蛋白潜在致敏性的细胞模型提供了一定的科学依据。The widely-used allergic diagnosis method depends on the binding of IgE, present in human patient sera. However, little correlation was demonstrated between serum immunoglobulin E（IgE） antibody level and the severity of clinical symptoms. Therefore, this study aimed to develop an in vitro cellular test system for evaluating potential allergenicity of foods via analysing the binding of mouse（Mus musculus） serum IgE to rat（Rattus norvegicus） basophilic leukaemia（RBL） cells and the degranulation of RBL-2H3 and RBL-1 cells. Female Balb/c mice （each 3-week-old） were orally sensitized 5 times with glycinin （Gly）, ovalbumin （OVA）, or potato acid phosphatase （PAP）. The sensitized serum pool that obtained after sensitization was utilized to analyze the binding of mouse IgE to RBL cells and mediator release assay. Additionally, elicitation of allergic reaction was assessed by measurement of vascular permeability to investigate the correlation between the degranulation of RBL cells and severity of allergic reaction. Results indicated that the capacity of RBL-2H3 cells to bind mouse IgE was significantly higher than that of RBL-1 cells （P〈0.05）. Good degranulation was induced in RBL-2H3 cell-line, while RBL-1 cell-line did the reverse. RBL-2H3 cell mediator release assay proved to be sensitive. Sensitized cells could be induced a maximal degranulation at a concentration of 10~100 ng/mL of allergens. This assay was also quite specific. Cells sensitized with sera from mouse with anti-OVA IgE did not degranulate following exposure to Gly. Furthermore, this assay had the ability to differentiate potential allergenicity of Gly, OVA and PAP. These functional data were found to be in line with the results of systemic anaphylaxis and demonstrated the ability of a protein to induce IgE-mediated allergic reactions. In conclusion, the RBL-2H3 cells, but not RBL-1 cells, is suitable for evaluating biological activity of different food allergens and form the basis of a useful model system for evaluat

关 键 词：RBL-2H3细胞 RBL-1细胞 食品过敏原 致敏活性 脱颗粒 

分 类 号：S852.651[农业科学—基础兽医学]