Flagellin Enhances the Immunogenicity of Pasteurella multocida Lipoprotein E Subunit Vaccine.

Yao-Chi Chung, Li-Ting Cheng, Chun-Yen Chu, Haroon Afzal, Thu-Dung Doan
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Abstract

Fowl cholera, caused by Pasteurella multocida infection, poses challenges for prevention because of its many serotypes. Bacterins are currently widely used for vaccination against fowl cholera, but protection is limited to homologous strains. Live attenuated vaccines of P. multocida provide some heterologous protection, but side effects are considerable. More recently, protein-based antigens are promising subunit vaccines when their low immunogenicity has been addressed with effective adjuvants. Bacterial flagellin has been widely considered a promising adjuvant for vaccines. In this study, we tested the adjutancy of flagellin in a subunit vaccine against P. multocida in a mice and chicken models. For vaccine formulation, the antigen fPlpE (P. multocida liporotein E) was combined with fFliC (Salmonella Typhimurium flagellin). The recombinant proteins of fPlpE and fFliC were successfully expressed using the Escherichia coli system as the expected sizes of 55 kDa and 70 kDa, respectively. The fFliC elicited strong expression levels of proinflammatory cytokine (IL-1β, IL-8, and IL-6) when stimulated in native chicken peripheral blood mononuclear cells. Immunization of mice and chickens with the subunit vaccines containing fFliC accelerated the antibody response. In the challenge tests, fFliC increased vaccine protective efficacy against the heterologous strain P. multocida A1 and highly virulent strain Chu01 in mice and chickens, respectively. These data indicated potential possibilities of using fFliC as an immunostimulant adjuvant in developing a subunit vaccine against fowl cholera.

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鞭毛蛋白能增强多杀性巴氏杆菌脂蛋白 E 亚基疫苗的免疫原性
鸡霍乱是由多杀性巴氏杆菌感染引起的,由于其血清型众多,给预防工作带来了挑战。目前,细菌素被广泛用于鸡霍乱疫苗接种,但其保护作用仅限于同源菌株。多杀性猪霍乱减毒活疫苗可提供一些异源保护,但副作用很大。最近,以蛋白质为基础的抗原在使用有效佐剂解决了低免疫原性的问题后,成为很有前景的亚单位疫苗。细菌鞭毛蛋白被广泛认为是一种很有前景的疫苗佐剂。在本研究中,我们在小鼠和鸡模型中测试了鞭毛蛋白在多杀性疟原虫亚单位疫苗中的辅助作用。在疫苗配制过程中,抗原 fPlpE(多杀性白喉杆菌脂溶血素 E)与 fFliC(鼠伤寒沙门氏菌鞭毛蛋白)相结合。fPlpE 和 fFliC 的重组蛋白在大肠杆菌系统中成功表达,预期大小分别为 55 kDa 和 70 kDa。在刺激原生鸡外周血单核细胞时,fFliC 能引起促炎细胞因子(IL-1β、IL-8 和 IL-6)的强烈表达。用含有 fFliC 的亚单位疫苗免疫小鼠和鸡可加速抗体反应。在挑战试验中,fFliC 分别提高了小鼠和鸡对异源株多杀性白喉杆菌 A1 和高毒力株 Chu01 的疫苗保护效力。这些数据表明,使用 fFliC 作为免疫增强佐剂开发鸡霍乱亚单位疫苗具有潜在的可能性。
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