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重组 Der P1 变应原衍生肽基疫苗在变应性小鼠模型中的变应原特异性免疫治疗。

Allergen-specific immunotherapy by recombinant Der P1 allergen-derived peptide-based vaccine in an allergic mouse model.

机构信息

Department of Molecular Genetics, Marvdasht Branch, Islamic Azad University, Marvdasht, Iran.

Immunology, Asthma and Allergy Research Institute (IAARI), Tehran University of Medical Sciences, Tehran, Iran.

出版信息

Immun Inflamm Dis. 2023 Jun;11(6):e878. doi: 10.1002/iid3.878.

Abstract

AIM

Increased IgE levels have made house dust mite allergens one of the most frequent causes of allergies worldwide. Treatment reduces the IgE antibodies and types two cytokines, namely interleukin-4 (IL-4) and IL-13. Although existing treatments significantly reduce IgE or IL-4/IL-13, they are very costly. This study aimed to construct a recombinant protein derived from rDer p1 peptides in the form of an immunotherapy approach and to measure the response of IgE and IgG antibodies.

METHODS

The proteins were isolated, purified, and evaluated using the SDS-PAGE and Bradford test and confirmed by using Western blot. To evaluate immunotherapy efficiency, 24 BALB/C mice were sensitized intraperitoneally with house dust mites (HDM) adsorbed to Aluminum hydroxide (Alum) and randomly divided into four groups of six: control sensitized, HDM extract, rDer p1, and DpTTDp vaccine. To immunization, four groups of random mice were each treated with phosphate-buffered saline, 100 μg of rDer p1 protein, DpTTDp, or HDM extract, every 3 days. Direct ELISA determined HDM-specific IgG and IgE subclasses. Data were analyzed in SPSS and Graph pad prism software. Values of p < .05 were considered significant.

RESULTS

After immunization of mice, the rDer P1 and recombinant vaccine like HDM extract increased IgG antibody titer and decreased IgE-dependent reactivity in allergic mice to rDer P1. Also, the levels of inflammatory IL-4 and IL-13 cytokines as allergic stimulants decreased.

CONCLUSION

The use of present available recombinant proteins is considered a viable, cost-effective, and long-term option for providing effective HDM allergy immunotherapy vaccines without side effects.

摘要

目的

IgE 水平的升高使屋尘螨过敏原成为全球最常见的过敏原因之一。治疗可降低 IgE 抗体和两种细胞因子,即白细胞介素-4(IL-4)和白细胞介素-13(IL-13)。尽管现有治疗方法可显著降低 IgE 或 IL-4/IL-13,但费用非常高。本研究旨在构建一种源自 rDer p1 肽的重组蛋白作为免疫疗法,并测量 IgE 和 IgG 抗体的反应。

方法

使用 SDS-PAGE 和 Bradford 试验分离、纯化和评估蛋白质,并使用 Western blot 进行验证。为了评估免疫治疗的效率,将 24 只 BALB/C 小鼠用吸附在氢氧化铝(Alum)上的屋尘螨(HDM)经腹腔内致敏,并随机分为四组,每组 6 只:对照致敏组、HDM 提取物组、rDer p1 组和 DpTTDp 疫苗组。为了免疫,随机将四组小鼠分别用磷酸盐缓冲盐水、100μg rDer p1 蛋白、DpTTDp 或 HDM 提取物处理,每 3 天一次。直接 ELISA 测定 HDM 特异性 IgG 和 IgE 亚类。数据在 SPSS 和 Graph pad prism 软件中进行分析。p<0.05 被认为具有统计学意义。

结果

在对小鼠进行免疫接种后,rDer P1 和重组疫苗(如 HDM 提取物)增加了过敏性小鼠对 rDer P1 的 IgG 抗体滴度,并降低了 IgE 依赖性反应。此外,作为过敏刺激物的炎症性 IL-4 和 IL-13 细胞因子水平降低。

结论

使用现有可用的重组蛋白被认为是一种可行、具有成本效益的长期选择,可提供有效的 HDM 过敏免疫治疗疫苗,而无副作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3994/10251762/3b8f2f853c06/IID3-11-e878-g003.jpg

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