Department of Molecular Medicine, Mayo Clinic, Rochester, MN 55905, USA.
Vaccine. 2011 Feb 11;29(8):1710-20. doi: 10.1016/j.vaccine.2010.12.020. Epub 2010 Dec 21.
Helicobacter pylori is a Gram-negative, spiral-shaped microorganism associated with acute and chronic gastritis, peptic ulcer, gastric cancer and gastric lymphomas in humans. H. pylori neutrophil-activating protein (NAP) is a major virulence factor playing a central role in pathogenesis of mucosal inflammation by immune cell attraction and Th1 cytokine response polarization. NAP is protective antigen and promising vaccine candidate against H. pylori infection. Here we present the development of measles virus (MV) vaccine strain encoding the NAP antigen. In order to facilitate the extracellular transport and detection, NAP was inserted in the human lambda immunoglobulin chain replacing a major part of the variable domain. We generated two MV vectors expressing secretory NAP forms: MV-lambda-NAP encoding the full-length constant lambda light chain domain and MV-s-NAP encoding only the N-terminus of the lambda light chain with the leader peptide. Immunization of MV permissive Ifnarko-CD46Ge transgenic mice by a single intraperitoneal injection of the NAP-expressing strains induced a robust, long-term humoral and cellular immune response against MV. Nine months post vaccination measles-neutralizing antibody titers were above the serum level considered protective for humans. Furthermore, all animals immunized with MV strains expressing the secretory NAP antigen developed strong humoral immunity against NAP, reaching titers >1:10,000 within 2-4 weeks. IFN-γ ELISpot assay confirmed that NAP-encoding MV vectors can also stimulate NAP-specific cell-mediated immunity. Our data demonstrate that MV is an excellent vector platform for expression of bacterial antigens and development of vaccines for H. pylori immunoprophylaxis in humans.
幽门螺杆菌是一种革兰氏阴性、螺旋形的微生物，与人类的急性和慢性胃炎、消化性溃疡、胃癌和胃淋巴瘤有关。幽门螺杆菌中性粒细胞激活蛋白（NAP）是一种主要的毒力因子，通过免疫细胞吸引和 Th1 细胞因子反应极化，在黏膜炎症的发病机制中起核心作用。NAP 是保护性抗原，也是针对幽门螺杆菌感染的有前途的疫苗候选物。在这里，我们介绍了编码 NAP 抗原的麻疹病毒（MV）疫苗株的开发。为了便于细胞外运输和检测，将 NAP 插入人 lambda 免疫球蛋白链中，取代可变结构域的大部分。我们生成了两种表达分泌型 NAP 形式的 MV 载体：编码全长恒定 lambda 轻链结构域的 MV-lambda-NAP 和仅编码 lambda 轻链 N 端的 MV-s-NAP，带有前导肽。通过单次腹腔内注射表达 NAP 的菌株，对 MV 许可的 Ifnarko-CD46Ge 转基因小鼠进行免疫，可诱导针对 MV 的强大、长期的体液和细胞免疫反应。接种疫苗 9 个月后，麻疹中和抗体滴度高于人类保护性血清水平。此外，所有用表达分泌型 NAP 抗原的 MV 株免疫的动物均针对 NAP 产生了强烈的体液免疫，在 2-4 周内达到 >1:10,000 的滴度。IFN-γ ELISpot 测定证实，编码 NAP 的 MV 载体还可以刺激 NAP 特异性细胞介导的免疫。我们的数据表明，MV 是表达细菌抗原和开发用于人类幽门螺杆菌免疫预防的疫苗的优秀载体平台。
