Laboratory of Molecular Biology of Microorganisms, Federal University of Alfenas, Alfenas, Brazil.
Department of Biology, Federal University of Lavras, Lavras, Brazil.
Front Immunol. 2021 Dec 21;12:778475. doi: 10.3389/fimmu.2021.778475. eCollection 2021.
spp. are Gram-negative, facultative intracellular bacteria that cause brucellosis in humans and animals. Currently available live attenuated vaccines against brucellosis still have drawbacks. Therefore, subunit vaccines, produced using epitope-based antigens, have the advantage of being safe, cost-effective and efficacious. Here, we identified small RNAs expressed during early infection with bone marrow-derived macrophages (BMDMs) and an apolipoprotein N-acyltransferase (Int) was identified as the putative target of the greatest expressed small RNA. Decreased expression of Int was observed during BMDM infection and the protein sequence was evaluated to rationally select a putative immunogenic epitope by immunoinformatic, which was explored as a vaccinal candidate. C57BL/6 mice were immunized and challenged with , showing lower recovery in the number of viable bacteria in the liver, spleen, and axillary lymph node and greater production of IgG and fractions when compared to non-vaccinated mice. The vaccinated and infected mice showed the increased expression of , , and IL-6 following expression of the anti-inflammatory genes and in the liver, justifying the reduction in the number and size of the observed granulomas. BMDMs stimulated with splenocyte supernatants from vaccinated and infected mice increase the CD86+ marker, as well as expressing greater amounts of iNOS and the consequent increase in NO production, suggesting an increase in the phagocytic and microbicidal capacity of these cells to eliminate the bacteria.
spp. 是革兰氏阴性、兼性细胞内细菌,可引起人类和动物的布鲁氏菌病。目前用于预防布鲁氏菌病的活减毒疫苗仍存在缺陷。因此,基于表位抗原的亚单位疫苗具有安全、经济有效和有效的优势。在这里,我们鉴定了在骨髓来源的巨噬细胞(BMDM)早期感染期间表达的小 RNA,并且鉴定了载脂蛋白 N-酰基转移酶(Int)作为表达最大的小 RNA 的假定靶标。在 BMDM 感染期间观察到 Int 的表达减少,并通过免疫信息学评估蛋白质序列以合理选择假定的免疫原性表位,将其作为候选疫苗进行探索。用 免疫 C57BL/6 小鼠并进行攻毒,与未接种疫苗的小鼠相比,在肝脏、脾脏和腋窝淋巴结中活细菌数量的恢复较低,并且 IgG 和分数的产生更高。与未接种疫苗的感染小鼠相比,接种疫苗和感染的小鼠在肝脏中抗炎基因 和 的表达后,显示出抗炎基因 和 的表达增加,IL-6 和 ,这证明了观察到的肉芽肿的数量和大小减少。用接种疫苗和感染小鼠的脾细胞上清液刺激 BMDM 增加 CD86+标志物的表达,以及表达更高水平的 iNOS 和随后增加的 NO 产生,表明这些细胞的吞噬和杀菌能力增强,以消除细菌。