| 引用本文: | 桑晶,康桦,王佳炜,陈舒怀,徐玲燕,王木兰,洪利娅.体外嗜碱性粒细胞激活试验法的建立[J].中国现代应用药学,2020,37(8):949-952. |
| SANG Jing,KANG Hua,WANG Jiawei,CHEN Shuhuai,XU Lingyan,WANG Mulan,HONG Liya.Establishment of Human Basophil Activation Method in Vitro[J].Chin J Mod Appl Pharm(中国现代应用药学),2020,37(8):949-952. |
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| 体外嗜碱性粒细胞激活试验法的建立 |
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桑晶1, 康桦1, 王佳炜2, 陈舒怀1, 徐玲燕1, 王木兰2, 洪利娅1
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1.浙江省食品药品检验研究院, 国家药品监督管理局化妆品动物替代试验技术重点实验室, 杭州 310004;2.正大青春宝药业有限公司, 杭州 310004
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| 摘要: |
| 目的 建立基于流式细胞仪检测的体外嗜碱性粒细胞激活试验（human basophil activation test，h-BAT）方法，并探索该方法用于Ⅰ型变态反应检查的可行性。方法 利用人源白血病嗜碱性粒细胞KU812在Ⅰ型变态反应过程中经阳性剂激活后，细胞特异性表面抗原CD63和CD203表达特异性增强的原理建立体外Ⅰ型变态反应模型，并同步用人组胺酶联免疫法进行验证。结果 确定以终浓度为2μg·mL-1的N-甲酰甲硫氨酰-亮氨酰-苯丙氨酸为阳性剂，孵育2 h作为流式方法检测嗜碱性粒细胞激活的阳性成立条件，细胞相对组胺释放百分比增高结果进一步确认了阳性成立条件。结论 h-BAT法有测试简便、评价客观等优点，是一种较好的过敏反应细胞模型，可以用于现有豚鼠试验的替代。 |
| 关键词: 嗜碱性粒细胞激活试验 过敏反应模型 替代方法 |
| DOI：10.13748/j.cnki.issn1007-7693.2020.08.010 |
| 分类号:R965.2 |
| 基金项目:浙江省药品监管系统科技计划项目（2020010）；浙江省食品药品监管系统科技计划项目（BH201701） |
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| Establishment of Human Basophil Activation Method in Vitro |
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SANG Jing1, KANG Hua1, WANG Jiawei2, CHEN Shuhuai1, XU Lingyan1, WANG Mulan2, HONG Liya1
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1.Zhejiang Institute for Food and Drug Control, NMPA Key Laboratory for Animal Alternative Testing Technology of Cosmetics, Hangzhou 310004, China;2.Chiatai Qingchunbao Pharmaceutical Co., Ltd., Hangzhou 310004, China
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| Abstract: |
| OBJECTIVE To establish human basophil activation test in vitro by flow cytometry, and to explore its feasibility of assessment of type I allergy reaction. METHODS By the theory of the specific expression of CD63 and CD203 in human basophil KU812 could be enhanced by positive agents, established in vitro model of type I allergy, which was verified by the result of human histamine enzyme-linked immunosorbent assay. RESULTS Human basophil activation test by flow cytometry was set up with positive agent of 2 μg·mL-1fMLP for 2 h of incubation. The results of increase of cell histamine release percentage confirmed the positive establishment condition at the same time. CONCLUSION Human basophil activation test in vitro is a good cell model of anaphylaxis with the advantages of relatively simple and objective. It is a good animal alternative test for sensitization test with guinea pigs. |
| Key words: human basophil activation test allergic model alternative method |
