Li Jianzhen, Yang Miao, Chen Bin, Wang Zhenhua, Cao Yuheng, Yang Yang, Zhang Mengwei, Zhang Dongmei, Ni Xueqin, Zeng Yan, Pan Kangcheng
Animal Microecology Institute, Department of Animal and Plant Quarantine, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China.
College of Animal Husbandry and Veterinary, Chengdu Agricultural College, Chengdu 611130, China.
Microorganisms. 2024 Feb 21;12(3):439. doi: 10.3390/microorganisms12030439.
, a probiotic bacterium with engineering potential, is widely used for the expression of exogenous proteins. In this study, we utilized the integrative plasmid pDG364 to integrate the hemagglutinin-neuraminidase () gene from Newcastle disease virus (NDV) into the genome of the 168 model strain. We successfully constructed a recombinant strain (designated RH) that displays a truncated antigen fragment on the surface of its spores and further evaluated its immunogenic effects in mice. Using ELISA, we quantified the levels of IgG in serum and secretory IgA (sIgA) in intestinal contents. The results revealed that the recombinant RH elicited robust specific mucosal and humoral immune responses in mice. Furthermore, RH demonstrated potential mucosal immune adjuvant properties by fostering the development of immune organs and augmenting the number of lymphocytes in the small intestinal villi. Additionally, the strain significantly upregulated the relative expression of inflammatory cytokines such as IL-1β, IL-6, IL-10, TNF-α, and IFN-γ in the small intestinal mucosa. In conclusion, the RH strain developed in this study exhibits promising mucosal immunogenic effects. It holds potential as a candidate for an anti-NDV mucosal subunit vaccine and offers a novel preventive strategy for the poultry industry against this disease.
作为一种具有工程潜力的益生菌,被广泛用于外源蛋白的表达。在本研究中,我们利用整合质粒pDG364将新城疫病毒(NDV)的血凝素神经氨酸酶()基因整合到168模式菌株的基因组中。我们成功构建了一种重组菌株(命名为RH),其孢子表面展示截短的抗原片段,并进一步评估了其在小鼠中的免疫原性效应。通过ELISA,我们定量了血清中IgG水平和肠内容物中分泌型IgA(sIgA)水平。结果显示,重组RH在小鼠中引发了强烈的特异性黏膜和体液免疫反应。此外,RH通过促进免疫器官发育和增加小肠绒毛中淋巴细胞数量,表现出潜在的黏膜免疫佐剂特性。另外,该菌株显著上调了小肠黏膜中炎症细胞因子如IL-1β、IL-6、IL-10、TNF-α和IFN-γ的相对表达。总之,本研究中构建的RH菌株表现出有前景的黏膜免疫原性效应。它有潜力作为抗NDV黏膜亚单位疫苗的候选物,并为家禽业针对该疾病提供一种新的预防策略。