一种潜在鼻病毒疫苗的反向疫苗学方法

S. Hudu, Saadatu Haruna Shinkafi, Shuaibu Umar, B. Makun
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摘要

背景:鼻病毒(RVs)是普通感冒最重要的病原,是儿童和成人慢性支气管炎、哮喘和慢性阻塞性肺疾病(COPD)急性加重的三分之二的原因。本研究旨在通过反向疫苗学方法设计一种能够诱导针对大多数RV的交叉反应抗体的泛血清型疫苗。方法:采用生物信息学分析方法对所有已知RV血清型的衣壳蛋白(VP1、VP2、VP3和VP4)进行表征,并预测潜在的免疫基序。选择并化学合成了所有RV-A或B血清型中共有的至少包含9个共聚物的保守基序。构建并体外克隆了理想菌株(HRV-74)、VP1、VP2、VP3和VP4 4个编码衣壳蛋白的全长标记基因。表达后,纯化的重组蛋白也被皮下注射到其他各组家兔。测定特异性免疫球蛋白M (IgM)和G (IgG)对多肽、蛋白和全病毒的反应和交叉反应性。结果:获得的抗肽抗体对不同RV菌株具有交叉中和活性。通过ELISA检测,合成肽的抗体对相应的重组蛋白和抗原不同的RV菌株表现出交叉反应性。结论:研究结果表明,RV衣壳蛋白保守区对应的肽具有较强的免疫原性;此外,研究结果表明,它们的组合对于扩展对变异rv的交叉保护至关重要。
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Reverse Vaccinology Approach for a Potential Rhinovirus Vaccine
Background: Rhinoviruses (RVs) represent the most important aetiological agents of the common cold and are responsible for about two-thirds of acute exacerbations of chronic bronchitis, asthma, and chronic obstructive pulmonary disease (COPD) in both children and adults. This study aimed to design a pan-serotypic vaccine capable of inducing cross-reactive antibodies against most of the RV by using a reverse Vaccinology approach. Methods: Bioinformatics analysis was carried out to characterise the capsid proteins (VP1, VP2, VP3, and VP4) of all known RV serotypes and to predict potential immune motifs. Conserved motifs consisting at least nine-mers common across all RV-A or B serotypes were selected and synthesized chemically. Four tagged full-length genes coding the capsid proteins of an ideal strain (HRV-74), VP1, VP2, VP3, and VP4 were constructed and cloned in vitro. Upon expression , the purified recombinant proteins were also administered subcutaneously to other groups of rabbits. The responses and cross-reactivity of the specific immunoglobulin M (IgM) and G (IgG) to the peptides, proteins, and whole viruses were measured. Results: The obtained anti-peptide antibodies exhibited a cross-neutralizing activity for different RV strains in vitro. Antibodies raised to the synthetic peptides exhibited cross-reactivity against the corresponding recombinant proteins and antigenically distinct RV strains coated on plates via ELISA assay. Conclusions: The study findings indicated that the peptides corresponding to the conserved region of the RV capsid proteins were potent immunogenic; moreover, the findings showed that their combination was crucial for extending the cross-protection against variant RVs.
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