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基于低变应原肽的沙拐枣花粉过敏疫苗的设计与评价。

Design and evaluation of a hypoallergenic peptide-based vaccine for Salsola kali allergy.

机构信息

Department of Immunology, School of Public Health, Tehran University of Medical Sciences, Iran; Immunology Asthma & Allergy Research Institute, Tehran University of Medical Sciences, Iran.

Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Iran; Department of Applied Biosciences and Biotechnology, Faculty of Science and Technology, Midlands State University (MSU), Zimbabwe.

出版信息

Int Immunopharmacol. 2019 Jan;66:62-68. doi: 10.1016/j.intimp.2018.10.037. Epub 2018 Nov 13.

Abstract

BACKGROUND

The Salsola kali (S. kali) pollen is one of the most important causes of allergic rhinitis in the deserts and semi-desert areas. Immunotherapy with allergen extracts remains the only available treatment addressing the underlying mechanism of allergy. However, given the low efficacy of this method, it is necessary to find more effective and alternative therapeutic interventions using molecular biology and bioinformatics tools. In this study, a hypoallergenic vaccine was designed on the basis of B-cell epitope approach for S. kali immunotherapy.

METHODS

Using the Immune Epitope Database (IEDB), a 35-mer peptide was selected and chemically conjugated to a keyhole limpet hemocyanin (KLH) molecule. Specific IgG and IgE from immunized BALB/c mice sera against the vaccine (Sal k 1-KLH), S. kali extract and the recombinant protein, rSal k 1, were measured using ELISA. Also, inhibition of IgE by mouse IgG was evaluated using an inhibitory ELISA. Finally, the IgE reactivity and T-cell reactivity of the designed vaccine were evaluated by dot blot assay and MTT assay.

RESULTS

Vaccination with the vaccine produced high levels of protective IgG in mice, which inhibited the binding of patients IgE to recombinant proteins. The result showed that the designed vaccine, unlike the recombinant protein and extract, did not induce T-cell lymphocytes response and also exhibited decreased IgE reactivity.

CONCLUSION

The designed vaccine can be considered as a promising candidate for therapeutic allergen-specific immunotherapy.

摘要

背景

盐角草(S. kali)花粉是沙漠和半沙漠地区引起过敏性鼻炎的最重要原因之一。过敏原提取物的免疫疗法仍然是针对过敏根本机制的唯一可用治疗方法。然而,鉴于这种方法的疗效较低,有必要使用分子生物学和生物信息学工具寻找更有效和替代的治疗干预措施。在这项研究中,基于 B 细胞表位方法设计了一种低过敏原疫苗,用于 S. kali 免疫疗法。

方法

使用免疫表位数据库(IEDB),选择一个 35 -mer 肽并将其化学偶联到钥孔血蓝蛋白(KLH)分子上。使用 ELISA 测量针对疫苗(Sal k 1-KLH)、S. kali 提取物和重组蛋白 rSal k 1 的免疫 BALB/c 小鼠血清中的特异性 IgG 和 IgE。还使用抑制 ELISA 评估了小鼠 IgG 对 IgE 的抑制作用。最后,通过斑点印迹法和 MTT 法评估设计疫苗的 IgE 反应性和 T 细胞反应性。

结果

疫苗接种在小鼠中产生了高水平的保护性 IgG,抑制了患者 IgE 与重组蛋白的结合。结果表明,与重组蛋白和提取物不同,设计的疫苗不会诱导 T 细胞淋巴细胞反应,并且还表现出降低的 IgE 反应性。

结论

设计的疫苗可以被认为是治疗性过敏原特异性免疫疗法的有前途的候选物。

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