Bisinotto R S, Filho J C Oliveira, Narbus C, Machado V S, Murray E, Bicalho R C
Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853.
Department of Entomology, College of Agriculture and Life Sciences, Cornell University, Ithaca, NY 14853.
J Dairy Sci. 2016 May;99(5):3765-3776. doi: 10.3168/jds.2015-10401.
Understanding the role of fimbrial subunits during bacterial adherence and the host's immunological response against anchorage proteins is critical for the development of strategies to prevent pathogens from thriving. The objectives of the present study were to locate fimbria-related proteins in the genome of Trueperella pyogenes (CP007519), define their importance for bacterial adherence, and evaluate the association between serum antibodies against fimbrial subunits and uterine health in dairy cows. Using a BLASTp search through the GenBank database, 4 putative clusters for fimbrial assembly were identified in the genome of T. pyogenes, namely FimA, FimC, FimE, and the novel major fimbriae FimJ. The fimbrial proteins FimA, FimC, FimE, and surface-anchored protein (SAP) were cloned into the pET 26b (+) vector, expressed in Escherichia coli BL21, and purified using affinity chromatography. Serum antibodies against FimA, FimC, FimE, and SAP were determined by ELISA on d 260±3 of gestation and at 2±1 and 35±3 d in milk (DIM) to assess the relationship between antigenicity against fimbrial proteins and parameters of uterine health. Antibodies against FimC and FimE were greater both pre- and postpartum in cows from which T. pyogenes was recovered by uterine flushing at 35±3 DIM, whereas T. pyogenes infection was not associated with differences in serum concentrations of FimA and SAP antibodies. Likewise, concentrations of FimC antibodies were consistently greater in cows diagnosed with clinical endometritis at 35±3 DIM compared with healthy counterparts. These results suggest that fimbrial proteins evaluated in the present study, particularly FimC and FimE, are important for maintenance of T. pyogenes in the uterus postpartum and development of uterine diseases in dairy cattle. Additional research is warranted to elucidate the mechanisms by which each fimbrial subunit contributes to the establishment of uterine diseases, evaluate its effect on fertility responses, and assess its relevance as a target for vaccine development.
了解菌毛亚基在细菌黏附过程中的作用以及宿主针对锚定蛋白的免疫反应,对于制定预防病原体滋生的策略至关重要。本研究的目的是在化脓隐秘杆菌(CP007519)基因组中定位菌毛相关蛋白,确定它们对细菌黏附的重要性,并评估奶牛血清中针对菌毛亚基的抗体与子宫健康之间的关联。通过在GenBank数据库中进行BLASTp搜索,在化脓隐秘杆菌基因组中鉴定出4个假定的菌毛组装簇,即FimA、FimC、FimE和新型主要菌毛FimJ。将菌毛蛋白FimA、FimC、FimE和表面锚定蛋白(SAP)克隆到pET 26b(+)载体中,在大肠杆菌BL21中表达,并使用亲和层析进行纯化。在妊娠260±3天以及产后2±1天和35±3天的乳中(DIM),通过ELISA测定针对FimA、FimC、FimE和SAP的血清抗体,以评估针对菌毛蛋白的抗原性与子宫健康参数之间的关系。在35±3 DIM通过子宫冲洗回收化脓隐秘杆菌的奶牛中,产前和产后针对FimC和FimE的抗体均较高,而化脓隐秘杆菌感染与FimA和SAP抗体的血清浓度差异无关。同样,在35±3 DIM被诊断为临床子宫内膜炎的奶牛中,FimC抗体浓度始终高于健康奶牛。这些结果表明,本研究中评估的菌毛蛋白,特别是FimC和FimE,对于产后化脓隐秘杆菌在子宫内的维持以及奶牛子宫疾病的发展很重要。有必要进行进一步的研究,以阐明每个菌毛亚基促成子宫疾病发生的机制,评估其对繁殖反应的影响,并评估其作为疫苗开发靶点的相关性。