Assessment of three human FcεRI-transfected RBL cell-lines for identifying IgE induced degranulation utilizing pea nut-allergic patient sera and peanut protein extract
Assessment of three human FcεRI-transfected RBL cell-lines for identifying IgE induced degranulation utilizing pea nut-allergic patient sera and peanut protein extract
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DOI：
10.1016/j.yrtph.2008.04.012
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发表时间：
2008-08-01
影响因子：
3.4
通讯作者：
Knippels, L. M. J.
中科院分区：
文献类型：
--
作者：
Ladics, G. S.;van Bilsen, J. H. M.;Knippels, L. M. J.
特异性IgE血清筛选研究用于研究蛋白质交叉反应性。这种非功能性免疫化学方法无法测量蛋白质的生物活性。因此，开发了使用用人Fc&RI转染的RBL细胞的测定。我们的目的是评价通过β-氨基己糖苷酶释放表达人Fc β RI的α-（RBL-hEI（a）-2B12和RBL-30/25细胞）或α-、β-和γ-亚基（RBL SX-38）的三种细胞系的脱粒。纯化的人IgE和血清衍生的多克隆IgE从花生过敏受试者后的挑战与抗IgE或花生蛋白提取物，分别被利用。使用纯化的IgE +抗人IgE，在所有三种中分别诱导了稳健的脱粒：RBL-30/25（84%）、hEI（a）-2B12（54%）、SX-38（94%）。对于一个花生过敏受试者+花生提取物，所有细胞系均发生良好的释放（分别为18%、40%、45%和65%）。对于来自其他三名花生过敏受试者的血清，RBL-hEI（a）-2B12细胞+花生提取物没有发生β-氨基己糖苷酶释放，而对于RBL-30/25和RBL SX 38细胞，仅来自一名受试者的血清诱导了良好的脱粒，分别为30%和60%。未观察到与强效食物过敏原（花生）一致的脱粒。该检测在安全性评估、预测值和用于评估蛋白质与过敏原的交叉反应性的再现性方面的实用性需要用其他蛋白质和充分表征的血清进行进一步研究。(c)2008年爱思唯尔公司All rights reserved.
Specific IgE sera screening studies are employed to investigate protein cross-reactivity. Such nonfunctional immunochemical methods cannot measure the biological activity of proteins. Therefore, an assay using RBL cells transfected with human Fc&RI was developed. Our objective was to evaluate the degranulation of three cell-lines expressing either the alpha-(RBL-hEI(a)-2B12 and RBL-30/25 cells) or alpha-, beta-, and gamma-subunits (RBL SX-38) of the human Fc epsilon RI by beta-hexosaminidase release. Purified human IgE and serumderived polyclonal IgE from peanut-allergic subjects following challenge with anti-IgE or peanut protein extract, respectively, were utilized. Robust degranulation was induced in all three: RBL-30/25 (84%), hEI(a)-2B12(54%), SX-38 (94%), respectively, using purified IgE + anti-human IgE. Good release (18%, 4045%, and 65%, respectively) occurred for one peanut-allergic subject + peanut extract with all cell-lines. With serum from three other peanut-allergic subjects, no beta-hexosaminidase release occurred with RBL-hEI(a)-2B12 cells + peanut extract, while only serum from one subject induced good degranulation, 30% and 60%, respectively, with RBL-30/25 and RBL SX 38 cells. Consistent degranulation with a potent food allergen (peanuts) was not observed. The assay's utility in safety assessment, predictive value and reproducibility for evaluating the cross-reactivity of proteins with allergens needs further investigation with additional proteins and well-characterized sera. (c) 2008 Elsevier Inc. All rights reserved.