Departamento de Medicina Veterinária Preventiva, Escola de Veterinária, Universidade Federal de Minas Gerais, Belo Horizonte 31270-010, Brazil; Departamento de Tecnologia e Inspeção de Produtos de Origem Animal, Escola de Veterinária, Universidade Federal de Minas Gerais, Belo Horizonte 31270-010, Brazil.
Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte 31270-901, Brazil.
J Dairy Sci. 2020 May;103(5):4588-4605. doi: 10.3168/jds.2019-17084. Epub 2020 Feb 26.
Staphylococcus aureus is one of the pathogens most frequently isolated from cases of mastitis worldwide. To decrease the effect of S. aureus mastitis in dairy farming, alternative strategies for controlling mastitis are needed that depend on a better knowledge of cow-to-cow variations in S. aureus antibody production. The present study sought to explore the diversity of S. aureus antibodies produced by dairy cows with a distinct mastitis history and vaccinated with a polyvalent mastitis vaccine. We obtained protein extracts from S. aureus isolates derived from persistent subclinical mastitis. Proteins were fractionated using 2-dimensional gel electrophoresis and Western blotting. Then, Western blotting membranes were exposed to sera from 24 dairy cows that had been divided into the following groups: vaccinated dairy cows that were infected with S. aureus, further subdivided according to whether they (a) remained infected by S. aureus or (b) recovered from the intramammary infection; unvaccinated dairy cows infected with S. aureus; and vaccinated healthy dairy cows with no history of S. aureus mastitis. Proteins found to be reactive by Western blot were identified by mass spectrometry (MALDI/TOF-TOF). Our most important finding was that FF ATP synthase subunit α, succinyl-diaminopimelate desuccinylase, and cysteinyl-tRNA synthetase were potential candidate proteins for the prevention of S. aureus mastitis. This study strengthens the notion that variations among animals should not be ignored and shows that the heterogeneity of antibody production against anti-staphylococcal antigens in animals may enable the identification of new immunotherapy targets.
金黄色葡萄球菌是世界范围内乳腺炎病例中最常分离到的病原体之一。为了降低奶牛乳腺炎中金黄色葡萄球菌的影响,需要依赖于对奶牛金黄色葡萄球菌抗体产生的个体差异的更好了解的替代控制乳腺炎策略。本研究旨在探索具有不同乳腺炎病史并接种多价乳腺炎疫苗的奶牛产生的金黄色葡萄球菌抗体的多样性。我们从持续亚临床乳腺炎中分离出金黄色葡萄球菌分离株获得蛋白提取物。使用二维凝胶电泳和 Western blotting 对蛋白质进行分级。然后,Western blotting 膜暴露于来自 24 头奶牛的血清中,这些奶牛分为以下组:接种了金黄色葡萄球菌的奶牛,根据它们(a)是否仍然感染金黄色葡萄球菌或(b)从乳房感染中恢复,进一步细分;感染了金黄色葡萄球菌的未接种奶牛;以及接种了金黄色葡萄球菌乳腺炎病史的健康奶牛。通过 Western blot 发现的反应蛋白通过质谱法(MALDI/TOF-TOF)进行鉴定。我们最重要的发现是,FF ATP 合酶亚基α、琥珀酰二氨基庚二酸去琥珀酸酶和半胱氨酰-tRNA 合成酶是预防金黄色葡萄球菌乳腺炎的潜在候选蛋白。这项研究加强了这样一种观念,即动物之间的差异不应被忽视,并表明针对抗葡萄球菌抗原的抗体产生的异质性可能使新的免疫治疗靶标得以识别。