In silico and in vivo Investigations of the Immunoreactivity of Klebsiella pneumoniae OmpA Protein as a Vaccine Candidate

Q2 Biochemistry, Genetics and Molecular Biology
Shahla Shahbazi, Farzad Badmasti, Mehri Habibi, Samira Sabzi, Narjes Noori Goodarzi, Mehdi Farokhi, Mohammad Reza Asadi Karam
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Abstract

Background: The growing threat of antibiotic resistance and Klebsiella pneumoniae infection in healthcare settings highlights the urgent need for innovative solutions, such as vaccines, to address these challenges. This study sought to assess the potential of using K. pneumoniae outer membrane protein A (OmpA) as a vaccine candidate through both in silico and in vivo analyses.

Methods: The study examined the OmpA protein sequence for subcellular localization, antigenicity, allergenicity, similarity to the human proteome, physicochemical properties, B-cell epitopes, MHC binding sites, tertiary structure predictions, molecular docking, and immune response simulations. The ompA gene was cloned into the pET-28a (+) vector, expressed, purified and confirmed using Western blotting analysis. IgG levels in the serum of the immunized mice were measured using ELISA with dilutions ranging from 1:100 to 1:6400, targeting recombinant outer membrane protein A (rOmpA) and K. pneumoniae ATCC 13883. The sensitivity and specificity of the ELISA method were also assessed.

Results: The bioinformatics analysis identified rOmpA as a promising vaccine candidate. The immunized group demonstrated significant production of specific total IgG antibodies against rOmpA and K. pneumoniae ATCC1 13883, as compared to the control group (p < 0.0001). The titers of antibodies produced in response to bacterial exposure did not show any significant difference when compared to the anti-rOmpA antibodies (p > 0.05). The ELISA test sensitivity was 1:3200, and the antibodies in the serum could accurately recognize K. pneumoniae cells.

Conclusion: This study is a significant advancement in the development of a potential vaccine against K. pneumoniae that relies on OmpA. Nevertheless, additional experimental analyses are required.

肺炎克雷伯菌 OmpA 蛋白作为候选疫苗的免疫反应性的硅学和体内研究。
背景:抗生素耐药性和肺炎克雷伯氏菌感染对医疗机构的威胁日益严重,这突出表明迫切需要疫苗等创新解决方案来应对这些挑战。本研究试图通过硅学和体内分析评估使用肺炎克雷伯菌 OmpA 作为候选疫苗的潜力:研究对 OmpA 蛋白序列的亚细胞定位、抗原性、致敏性、与人类蛋白质组的相似性、理化性质、B 细胞表位、MHC 结合位点、三级结构预测、分子对接和免疫反应模拟进行了检查。将 ompA 基因克隆到 pET-28a (+) 载体中,进行表达、纯化并通过 Western 印迹分析加以确认。免疫小鼠血清中的 IgG 含量是用 ELISA 法测定的,稀释度从 1:100 到 1:6400,靶标是 rOmpA 和 K. pneumoniae ATCC 13883。同时还评估了 ELISA 方法的灵敏度和特异性:结果:生物信息学分析发现 rOmpA 是一种很有前景的候选疫苗。与对照组相比,免疫组明显产生了针对 rOmpA 和肺炎克氏菌 ATCC1 13883 的特异性总 IgG 抗体(p < 0.0001)。与抗 rOmpA 抗体相比,细菌暴露后产生的抗体滴度没有明显差异(p > 0.05)。ELISA 试验的灵敏度为 1:3200,血清中的抗体能准确识别肺炎克氏菌细胞:结论:这项研究在开发依赖 OmpA 的潜在肺炎克氏菌疫苗方面取得了重大进展。尽管如此,还需要进行更多的实验分析。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Iranian Biomedical Journal
Iranian Biomedical Journal Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (all)
CiteScore
3.20
自引率
0.00%
发文量
42
审稿时长
8 weeks
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