外源蛋白潜在致敏性不同方法评价分析
Different methods to evaluate potential allergenicity of foreign allergens
-
摘要: 目的利用动物模型和生物信息学分析方法对3种外源蛋白(Cry1Ab/Ac、LRP和HSA)的潜在致敏性进行分析。方法动物实验将32只3周龄SPF级雌性Balb/c小鼠分为4组,对照组及Cry1Ab/Ac组、LRP组、HSA组分别以经口灌胃途径给予生理盐水及外源蛋白Cry1Ab/Ac、LRP、HSA,ELISA法测定血清中特异性IgE水平和血浆中组胺水平;利用生物信息学分析方法采用过敏原在线数据库将3种外源蛋白的氨基酸序列与已知过敏原进行对比,判断3种外源蛋白的潜在致敏性。结果动物模型实验结果显示,Cry1Ab/Ac组、LRP组、HSA组小鼠IgE抗体水平分别为(3.83±0.41)、(3.00±0.81)、(3.50±0.55),均高于对照组小鼠的(1.67±0.58),差异均有统计学意义(P<0.05);对照组、Cry1Ab/Ac组、LRP组、HSA组小鼠组胺水平分别为(976.46±179.55)、(1 308.56±465.17)、(985.89±453.54)、(1 149.61±200.97),差异均无统计学意义(P>0.05);生物信息学分析结果显示,HSA和LRP与已知食品过敏原存在同源性,可能存在潜在致敏性,而Cry1Ab/Ac未发现与已知食品过敏原存在同源性。结论Cry1Ab/Ac、LRP和HSA 3种外源蛋白均具有较强的免疫原性,其中 LRP和HSA具有潜在致敏性。Abstract: Objective To analyse the potential allergenicity of 3 foreign allergens(insecticidal crystal proteinCry1Ab/Ac),lysine-rich proteinLRP,and human serum albuminHSA)using animal model in vivo and bioinformatics analysis. Methods Amino acid sequences of the three foreign proteins were compared with those of the known allergens using online database.Female Balb/c mice were orally exposed to the three foreign proteins.Protein-specific IgE antibodies in serum and plasma histamine levels were detected with enzyme-linked immunosorbent assay(ELISA). Results The results of bioinformatics analysis showed that HSA and LRP matched with the known food allergens and had the potential allergenicity,but Cry1Ab/Ac didn't match with the known food allergens.The results of animal model showed that all of the 3 foreign proteins were potentially allergenic.Compared with negative control group,the 3 foreign proteins could increase the level of IgE.The results of histamine detection showed that the three foreign proteins did not significantly increase the histamine level. Conclusion Cry1Ab/Ac,LRP and HSA have strong immunogenicity and LRP and HSA have potential allergenicity.
下载: