Esmaeili Zahra, Kamal Shahsavar Sara, Ariannejad Hamid, Hajinajaf Nima, Menbari Shaho, Ghazvini Kiarash
Department of Microbiology and Virology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran; Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
Institute of Biotechnology, Ferdowsi University of Mashhad, Iran.
Microb Pathog. 2025 Jul;204:107613. doi: 10.1016/j.micpath.2025.107613. Epub 2025 Apr 17.
Helicobacter pylori (H.pylori) is considered to be the most important gastrointestinal pathogen causing gastritis, gastric ulcers and even gastric cancer. The treatment of these infections has failed due to the rapidly increasing antibiotic resistance to standard treatment regimens and the lack of an effective vaccine. This study investigates the production and therapeutic potential of Immunoglobulin Y (IgY) antibodies targeting key epitopes of the H. pylori UreB recombinant protein. Given the increasing challenge of antibiotic resistance in H. pylori treatment, this research underscores the necessity for alternative therapeutic strategies. A specific region of the UreB gene, containing critical immunogenic epitopes, was amplified using Polymerase Chain Reaction (PCR) and cloned into the pET32b vector. The recombinant plasmid was expressed in Escherichia coli BL21 (DE3), and the UreB protein was purified via Ni-NTA affinity chromatography, confirmed by SDS-PAGE and Western blot analysis. Hens were immunized with the recombinant UreB protein, resulting in the generation of specific IgY antibodies. The purified IgY-UreB antibodies exhibited a remarkable reduction in urease activity by 84.53 % at a concentration of 10 mg/mL, effectively neutralizing this critical virulence factor. Additionally, in vitro assays demonstrated that IgY-UreB antibodies significantly inhibited the growth of H. pylori at a concentration of 5 mg/mL. These findings highlight the potential of IgY as a viable alternative to traditional antibiotic therapies, particularly in the context of rising antibiotic resistance. This study paves the way for the development of innovative immunotherapeutic strategies that may improve treatment outcomes for H. pylori infections.
幽门螺杆菌(H.pylori)被认为是导致胃炎、胃溃疡甚至胃癌的最重要的胃肠道病原体。由于对标准治疗方案的抗生素耐药性迅速增加以及缺乏有效的疫苗,这些感染的治疗已经失败。本研究调查了针对幽门螺杆菌尿素酶B(UreB)重组蛋白关键表位的免疫球蛋白Y(IgY)抗体的产生及其治疗潜力。鉴于幽门螺杆菌治疗中抗生素耐药性日益增加的挑战,本研究强调了替代治疗策略的必要性。使用聚合酶链反应(PCR)扩增UreB基因的一个包含关键免疫原性表位的特定区域,并将其克隆到pET32b载体中。重组质粒在大肠杆菌BL21(DE3)中表达,通过镍-亚氨基二乙酸(Ni-NTA)亲和层析纯化UreB蛋白,经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白质免疫印迹分析确认。用重组UreB蛋白免疫母鸡,产生特异性IgY抗体。纯化的IgY-UreB抗体在浓度为10mg/mL时脲酶活性显著降低84.53%,有效中和了这一关键毒力因子。此外,体外试验表明,IgY-UreB抗体在浓度为5mg/mL时显著抑制幽门螺杆菌的生长。这些发现突出了IgY作为传统抗生素疗法可行替代方案的潜力,尤其是在抗生素耐药性不断上升的情况下。本研究为开发可能改善幽门螺杆菌感染治疗效果的创新免疫治疗策略铺平了道路。