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首页> 外文期刊>Infection and immunity >Antigens of Brucella abortus S19 immunodominant for bovine lymphocytes as identified by one- and two-dimensional cellular immunoblotting.
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Antigens of Brucella abortus S19 immunodominant for bovine lymphocytes as identified by one- and two-dimensional cellular immunoblotting.

机译：一维和二维细胞免疫印迹法鉴定了对牛淋巴细胞具有免疫力的流产布鲁氏菌S19抗原。

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Cellular immune responses are influential for protection against intracellular bacteria such as brucellae. Therefore, identification of Brucella abortus antigens that activate primed bovine lymphocytes is fundamental for discerning the breadth of cellular response in bovine brucellosis. Potentially antigenic components of B. abortus S19 were isolated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by nitrocellulose blotting. Specific one-dimensional blot segments induced proliferation of peripheral blood lymphocytes from all 25 of the vaccinated cattle tested and were defined as immunodominant. Individual proteins that stimulated lymphocyte proliferation were further characterized by two-dimensional cellular immunoblotting by two different approaches. Individual one-dimensional stimulatory blot segments were eluted, concentrated, and then subjected to two-dimensional cellular immunoblotting. Alternatively, entire two-dimensional gels containing all of the B. abortus components were blotted and nitrocellulose sections containing individual proteins were assayed for lymphocyte activation. Thirty-eight Brucella proteins that induced lymphocyte proliferation were resolved by both procedures. Phenotypic analysis of the proliferating cell population demonstrated the presence of CD4+, CD8+, and immunoglobulin M+ lymphocytes. Two immunogenic proteins, 12 and 31 kDa, identified by two-dimensional cellular immunoblotting, were subjected to partial N-terminal amino acid analysis. The 12-kDa protein was within the area of greatest lymphocyte proliferation, while the 31-kDa protein was chosen for comparison with a 31-kDa protein previously reported by others. A search of the National Biomedical Research Foundation protein data bank showed that the sequences were not homologous with other known proteins. Identification of Brucella proteins immunogenic for bovine lymphocytes provides an important step in distinguishing the various proteins involved in pathogenicity and/or disease resistance.
机译：细胞免疫应答对于保护细胞内细菌如布鲁氏菌有影响。因此，鉴定能激活牛淋巴细胞的流产布鲁氏菌抗原对于辨别牛布鲁氏菌病中细胞反应的广度至关重要。流产双歧杆菌S19的潜在抗原成分通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳，然后进行硝酸纤维素印迹进行分离。特定的一维印迹片段诱导了来自所有25只经免疫接种的牛的外周血淋巴细胞的增殖，并被定义为免疫显性的。通过两种不同方法通过二维细胞免疫印迹进一步表征了刺激淋巴细胞增殖的单个蛋白质。洗脱单个一维刺激印迹片段，浓缩，然后进行二维细胞免疫印迹。或者，将包含所有流产双歧杆菌成分的整个二维凝胶印迹，并分析包含单个蛋白质的硝酸纤维素切片的淋巴细胞活化作用。两种方法均能消除诱导淋巴细胞增殖的38种布鲁氏菌蛋白。对增殖细胞群的表型分析表明存在CD4 +，CD8 +和免疫球蛋白M +淋巴细胞。通过二维细胞免疫印迹法鉴定的两种免疫原性蛋白（12和31 kDa）经过部分N末端氨基酸分析。 12 kDa蛋白位于淋巴细胞增殖最大的区域内，而选择31 kDa蛋白与其他人先前报道的31 kDa蛋白进行比较。对美国国家生物医学研究基金会蛋白质数据库的搜索显示，该序列与其他已知蛋白质不同源。对牛淋巴细胞具有免疫原性的布鲁氏菌蛋白质的鉴定为区分涉及致病性和/或疾病抗性的各种蛋白质提供了重要的步骤。

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