Nandre Rahul M, Ruan Xiaosai, Duan Qiangde, Sack David A, Zhang Weiping
Department of Diagnostic Medicine/Pathobiology, Kansas State University College of Veterinary Medicine, Manhattan, KS, USA.
Department of International Health, Johns Hopkins University Bloomberg School of Public Health, Baltimore, MD, USA.
Vaccine. 2016 Jun 30;34(31):3620-5. doi: 10.1016/j.vaccine.2016.04.003. Epub 2016 May 24.
Diarrhea continues to be a leading cause of death in children younger than 5 years in developing countries. Enterotoxigenic Escherichia coli (ETEC) is a leading bacterial cause of children's diarrhea and travelers' diarrhea. ETEC bacteria initiate diarrheal disease by attaching to host receptors at epithelial cells and colonizing in small intestine. Therefore, preventing ETEC attachment has been considered the first line of defense against ETEC diarrhea. However, developing vaccines effectively against ETEC bacterial attachment encounters challenge because ETEC strains produce over 23 immunologically heterogeneous adhesins. In this study, we applied MEFA (multiepitope fusion antigen) approach to integrate epitopes from adhesin tips or adhesive subunits of CFA/I, CS1, CS2, CS3, CS4, CS5, CS6, CS21 and EtpA adhesins and to construct an adhesin tip MEFA peptide. We then examined immunogenicity of this tip MEFA in mouse immunization, and assessed potential application of this tip MEFA for ETEC vaccine development. Data showed that mice intraperitoneally immunized with this adhesin tip MEFA developed IgG antibody responses to all nine ETEC adhesins. Moreover, ETEC and E. coli bacteria expressing these nine adhesins, after incubation with serum of the immunized mice, exhibited significant reduction in attachment to Caco-2 cells. These results indicated that anti-adhesin antibodies induced by this adhesin tip MEFA blocked adherence of the most important ETEC adhesins, suggesting this multivalent tip MEFA may be useful for developing a broadly protective anti-adhesin vaccine against ETEC diarrhea.
腹泻仍然是发展中国家5岁以下儿童死亡的主要原因。产肠毒素大肠杆菌(ETEC)是儿童腹泻和旅行者腹泻的主要细菌病因。ETEC细菌通过附着于上皮细胞的宿主受体并在小肠中定殖来引发腹泻病。因此,预防ETEC附着一直被视为抵御ETEC腹泻的第一道防线。然而,开发有效的抗ETEC细菌附着疫苗面临挑战,因为ETEC菌株产生超过23种免疫异质性粘附素。在本研究中,我们应用多表位融合抗原(MEFA)方法整合来自CFA/I、CS1、CS2、CS3、CS4、CS5、CS6、CS21和EtpA粘附素的粘附素尖端或粘附亚基的表位,并构建一种粘附素尖端MEFA肽。然后我们检测了这种尖端MEFA在小鼠免疫中的免疫原性,并评估了这种尖端MEFA在ETEC疫苗开发中的潜在应用。数据显示,用这种粘附素尖端MEFA进行腹腔免疫的小鼠对所有九种ETEC粘附素产生了IgG抗体反应。此外,表达这九种粘附素的ETEC和大肠杆菌在与免疫小鼠的血清孵育后,对Caco-2细胞的附着显著减少。这些结果表明,这种粘附素尖端MEFA诱导的抗粘附素抗体阻断了最重要的ETEC粘附素的粘附,表明这种多价尖端MEFA可能有助于开发一种针对ETEC腹泻的具有广泛保护作用的抗粘附素疫苗。