Han Lichao, Ji Xingzhao, Fan Shihong, Shen Jirao, Liang Bin, Li Zhenjun
Department of Traditional Chinese Medicine, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, China.
Chinese Center for Disease Control and Prevention, State Key Laboratory of Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, 155 Changbai Road Changping District, 102206, Beiing, People's Republic of China.
Trop Dis Travel Med Vaccines. 2024 Oct 15;10(1):21. doi: 10.1186/s40794-024-00229-w.
Nocardia is emerging as a common and easily neglected cause of both healthcare- and occupation-associated infections worldwide, however, human vaccines for Nocardia prevention are not yet available. In this study, we aimed to evaluate the immunoprotective effect of the NFA47630 protein, a secreted protein abundant in the N. farcinica IFM10152 supernatant.
Conservation and characteristics of nfa47630 were analyzed by PCR and bioinformatics. Then recombinant NFA47630 protein was cloned, expressed and purified for further antigenicity analysis. Subsequently, the ability to activate innate immunity was evaluated by examining the phosphorylation status of the MAPK signaling pathway and cytokine levels. Finally, the protective effect was evaluated on rNFA47630-immunized mice.
nfa47630 was conserved in N. farcinica strains with good antigenicity. The rNFA47630 protein was expressed under the optimal conditions of 0.2 mM IPTG, 28 °C, and it can be recognized by anti-N. farcinica and anti-N. cyriacigeorgica sera, but not anti-N. asteroids, anti-N. brasiliensis, anti-N. nova and anti-Mycobacterium bovis sera. It can upregulate the phosphorylation status of ERK, JNK, P38 and the cytokine levels of TNF-α, IL-10, IL-12, and IFN-γ. In addition, mice immunized with rNFA47630 protein exhibited higher antibody titers, greater bacterial clearance ability, milder organ infection, and higher survival rates than PBS-immunized mice.
Our data demonstrate that NFA47630 is a potential vaccine candidate for defending against N. farcinica infection.
诺卡菌正成为全球医疗保健相关和职业相关感染中一种常见且易被忽视的病因，然而，用于预防诺卡菌的人类疫苗尚未问世。在本研究中，我们旨在评估NFA47630蛋白的免疫保护作用，该蛋白是嗜皮诺卡菌IFM10152上清液中丰富的一种分泌蛋白。
通过PCR和生物信息学分析nfa47630的保守性和特征。然后克隆、表达和纯化重组NFA47630蛋白，用于进一步的抗原性分析。随后，通过检测MAPK信号通路的磷酸化状态和细胞因子水平来评估激活先天免疫的能力。最后，对用rNFA47630免疫的小鼠评估其保护作用。
nfa47630在嗜皮诺卡菌菌株中保守，具有良好的抗原性。rNFA47630蛋白在0.2 mM IPTG、28°C的最佳条件下表达，可被抗嗜皮诺卡菌和抗乔治西里亚克诺卡菌血清识别，但不能被抗星状诺卡菌、抗巴西诺卡菌、抗新星诺卡菌和抗牛分枝杆菌血清识别。它可以上调ERK、JNK、P38的磷酸化状态以及TNF-α、IL-10、IL-12和IFN-γ的细胞因子水平。此外，用rNFA47630蛋白免疫的小鼠比用PBS免疫的小鼠表现出更高的抗体滴度、更强的细菌清除能力、更轻的器官感染和更高的存活率。
我们的数据表明，NFA47630是一种潜在的抗嗜皮诺卡菌感染疫苗候选物。
