T CELL RESPONSE TO AA IN MICE

T细胞伴放线放线杆菌的新用途 伴放线放线杆菌（Aa）a革兰氏阴性 细菌，与局部青少年密切相关 牙周炎（LJP），其影响约0.5%的美国人。 孩子 LJP可通过抗生素治疗成功控制， 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， 治疗可能提供控制LJP的更好方法。 在 在动物研究中，T细胞已被证明在 保护宿主免受Aa的侵害，但目前知之甚少 引起T细胞反应的特异性Aa蛋白。 到 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的小鼠的淋巴结。到 确定Aa蛋白的大小， 杂交瘤，使用“T细胞蛋白质印迹”。 有趣的是，6 迄今为止测试的11种T细胞杂交瘤显示出对 相对高分子量的蛋白质。 以来 白细胞毒素，由其产生的主要病毒因子， 细菌，是在这个大小范围内的少数蛋白质之一，我们 怀疑它可能是相关抗原。 因此这些 测试杂交瘤对蛋白质提取物的反应性 由IP-2和AAM 68（一种同基因白细胞毒素）制备 敲除菌株）; 5/6个T细胞杂交瘤被 对白细胞毒素有特异性 我们首先定义了 一种抗原，激发一个强大的，潜在的保护性T细胞 反应 此外，为了消除姐妹克隆， T细胞受体B链连接区的序列是 测序 对Aa的反应似乎相当异质， 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活性。 T细胞 杂交瘤以前从未用于Aa的研究，它们 提供了一个重要的新工具， 反应和疾病的病因，这一重要的 牙周病原体 这项工作得到了国家 卫生研究院，赠款DE 00152和DE 10731。

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.