T CELL RESPONSE TO AA IN MICE

伴放线T细胞放线杆菌的新用途 伴生放线杆菌(Aa)革兰氏阴性杆菌 细菌，与局部性青少年密切相关 牙周炎(LJP)影响着美国约0.5%的人口。 孩子们。通过抗生素治疗成功地治疗了LJP 手术，但对牙周组织的广泛损害往往 发生在检测或治疗之前。因此，基于免疫的 治疗可能提供一种更好的控制LJP的方法。在……里面 动物研究表明，T细胞在 保护主机免受AA攻击，但目前知之甚少 关于能引起T细胞反应的特定AA蛋白。至 为了解决这个问题，我们已经产生了24个T细胞杂交瘤 使用口服接种AA的小鼠的淋巴。至 确定它们所识别的AA蛋白的大小 杂交瘤，“T细胞西部片”被使用。有趣的是，其中6个 到目前为止测试的11个T细胞杂交瘤显示对An的反应性 相对分子质量较高的AA蛋白。自.以来 白毒素，由它产生的主要致病因子 细菌是这个大小范围内为数不多的蛋白质之一，我们 怀疑它可能是相关的抗原。因此，这些 杂交瘤细胞对蛋白质提取物的反应性测试 从IP-2和AAM68(一种同源白毒素)制备 我们产生的敲除菌株)；5/6 T细胞杂交瘤 确实是白毒素的特异体。因此，我们定义了第一个 AA抗原，可诱导强大的、具有潜在保护性的T细胞 回应。此外，为了消除姐妹克隆， T细胞受体B链连接区的序列如下 已排序。对AA的反应看起来相当不同， 代表了许多不同的VB基因片段；这表明 AA在小鼠体内没有超抗原性。T细胞 杂交瘤以前从未被用于研究再生障碍性贫血，他们 提供了一个重要的新工具，有助于解剖免疫 对这一重要事件的反应和疾病病因 牙周病原体。这项工作得到了国家 卫生研究所，赠款DE00152和DE10731。

Novel use of T cell Actinobacillus actinomycetemcomitans Actinobacillus actinomycetemcomitans (Aa) a gram-negative bacterium, is strongly associated with Localized Juvenile periodontitis (LJP) which affects approximately 0.5% of the U.S. children. LJP is successfully managed by antibiotic therapy and surgery, but extensive damage to the periodontium has often occurred before detection or treatment. Thus an immune based therapy may provide a better method of controlling LJP. In animal studies, T cells have been shown to be important in protecting the host against Aa, but little is currently known about the specific Aa proteins that elicit a T cell response. To address this question, we have generated 24 T cell hybridomas using lymph nodes from mice orally inoculated with Aa. To determine the sizes of the Aa proteins recognized by these hybridomas, "T cell Westerns" were employed. Interestingly, 6 of the 11 T cell hybridomas tested to date showed reactivity to an Aa protein of relatively high molecular weight. Since leukotoxin, the primary vi rulence factor produced by this bacterium, is one of only a few proteins in that size range, we suspected that it might be the relevant antigen. Thus, these hybridomas were tested for reactivity to protein extracts prepared from IP-2 and from AAM68 (an isogenic leukotoxin knockout strain that we generated); 5/6 T cell hybridomas were indeed specific for leukotoxin. Thus, we have defined the first Aa antigen that elicits a strong, potentially protective, T cell response. In addition, in order to eliminate sister clones, the sequences of the T cell receptor B chain junctional regions were sequenced. The response to Aa appears quite heterogeneous and many difference VB gene segments are represented; this suggests that Aa has no super-antigenic activity in mice. T cell hybridomas have never before been used in the study of Aa, they provide an important new tool useful in dissecting the immune response and disease etiology seen with this important periodontal pathogen. This work was supported by the National Institutes of Health, grants DE00152 and DE10731.