Fang Luis, Martínez Dalgys, Meza-Torres Catherine, Pereira-Sanandrés Nicole, Moreno-Woo Ana, Garavito Gloria, Egea Eduardo
Universidad del Norte. División de Ciencias de la Salud, Departamento de Medicina, Barranquilla, Colombia.
Universidad de Cartagena. Instituto de Investigaciones Inmunológicas, Cartagena, Colombia.
Rev Alerg Mex. 2024 Feb 1;71(1):68. doi: 10.29262/ram.v71i1.1366.
The objective of the present study was to design a multi-epitope protein from and APD allergens and to evaluate its IgE reactivity preliminarily.
Using computational tools, a molecule containing multiple "T" epitopes of allergens derived from and APD was designed "" This multi-epitope protein (MP1) was expressed using an system and purified by affinity chromatography using Ni-NTA agarose. Anti-MP1 and anti-HDM extract IgE reactivity was evaluated by Dot-Blot and indirect ELISA from sera of HDM-allergic patients and non-allergic individuals from Barranquilla-Colombia. Allergic individuals had a positive skin test to a standardized battery of inhaled allergens (EUROLINE - Ref: DP 3704-1601-1 E) and mite- specific IgE.
Multi-epitope (MP1) protein was expressed and purified with high purity. Dot-Blot result showed that all sera from allergic patients showed lower IgE reactivity to MP1 compared to HDM extract. By ELISA, significantly lower concentrations of anti-MP1 IgE (Median: 270.86 ng/ml; IQR: 90.3) were observed in contrast to anti-HDM IgE levels (Median: 988.5 ng/ml; IQR: 1117.6) in sera of patients allergic to HDM.
A protein composed of multiple epitopes of and HDM allergens was designed, expressed, and purified. Preliminary Dot-Blot results suggest that this molecule shows hypoallergenic properties with very low IgE reactivity compared to mite extract. Further functional studies are needed to understand better the immune response induced by this molecule.
本研究的目的是从和APD过敏原设计一种多表位蛋白，并初步评估其IgE反应性。
使用计算工具，设计了一种包含来自和APD的过敏原多个“T”表位的分子“” 这种多表位蛋白（MP1）使用系统表达，并通过使用Ni-NTA琼脂糖的亲和色谱法纯化。通过斑点印迹和间接ELISA法评估来自哥伦比亚巴兰基亚的HDM过敏患者和非过敏个体血清中抗MP1和抗HDM提取物的IgE反应性。过敏个体对标准化吸入过敏原组合（EUROLINE - 参考编号：DP 3704-1601-1 E）和螨特异性IgE的皮肤试验呈阳性。
多表位（MP1）蛋白得以表达并高纯度纯化。斑点印迹结果显示，与HDM提取物相比，所有过敏患者血清对MP1的IgE反应性较低。通过ELISA法，在对HDM过敏的患者血清中，观察到抗MP1 IgE的浓度显著较低（中位数：270.86 ng/ml；四分位间距：90.3），而抗HDM IgE水平（中位数：988.5 ng/ml；四分位间距：1117.6）。
设计、表达并纯化了一种由和HDM过敏原的多个表位组成的蛋白。初步斑点印迹结果表明，与螨提取物相比，该分子具有低过敏性，IgE反应性极低。需要进一步的功能研究以更好地了解该分子诱导的免疫反应。
