T CELLS/CYTOKINES FOR B CELL RESPONSES IN ORAL DISEASE

T 细胞/细胞因子对口腔疾病中 B 细胞的反应

• 批准号: 6270278
• 负责人: HIROSHI KIYONO
• 金额: $ 22.56万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-12-01 至 2000-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6270278
• 关键词: B lymphocyte; Bacteroides gingivalis; antibody formation; bacterial antigens; bactericidal immunity; cytokine; enzyme linked immunosorbent assay; fibroblasts; flow cytometry; gene rearrangement; gingiva; helper T lymphocyte; human tissue; immunoglobulin genes; immunopathology; in situ hybridization; inflammation; interferon gamma; interleukin 2; interleukin 4; interleukin 5; interleukin 6; laboratory mouse; lymphocyte proliferation; molecular cloning; monocyte; oral bacteria; periodontium disorder; polymerase chain reaction; stress proteins; superantigens; tissue /cell culture

Our past work on this grant directly addressed immune aspects of Periodontal Disease (PD) by development of methods for isolation of gingival mononuclear cells (GMC) from the disease site. We have shown that large numbers of immunoglobulin (Ig) secreting, spot forming cells (SFC) occur, which are mainly restricted to IgG-subclass (IgG1>IgG2>IgG3=IgG4) and less so to IgA-subclass (IgA1>IgA2) responses. This unique system has allowed us to determine the degree of antigen- specific SFC responses in PD, and we showed that Porphyromonas gingivalis fimbriae and LPS are major and minor components of these IgG- and IgA- subclass responses, respectively. Thus, our work suggests that the predominant IgG- and IgA-plasma cell responses seen in PD have both polyclonal and antigen-specific components. This has logically led to an analysis of cytokines present in GMC supernatants (sups), which would account for these unique profiles of B cell responses. Our studies have now shown that IL-6 is a major terminal differentiation factor in GMC sups, together with IL-5. Of importance to this proposal is the presence of a potentially novel cytokine which induces IL-6 receptor (IL-6R) expression on human lymphocytes. We plan to continue these exciting studies by addressing the precise pattern of cytokines present in individual cell types in GMC which account for B cell and plasma cell responses. We will focus on IL-2, IL-4, IL-5, IL-6 and IFN-gamma and use both PCR and in situ hybridization for these cytokines in both immune (T helper (Th) cells subsets, B cells, monocytes/macrophages) and nonimmune (fibroblasts and epithelial cells) cells from tissues of patients with PD. Our preliminary studies suggest that gingival Th cells may be induced, in part, by the presence of P. gingivalis fimbriae acting as a superantigen and by heat shock proteins (hsps). We plan to assess CD4+ Th cells from GMC for responses to these antigens and to determine if hsps induce Th1-type (IFN-gamma and IL-2) and fimbriae (superantigen) elicits mainly Th2 (IL-4, IL-5 and IL-6) responses. By specific derivation of cloned CD4+ Th cells, we can determine the relative contribution of Th1 and Th2 cells for inflammation and B cell responses, respectively. The potential role of TGF-beta and IL-4 in switches to IgG- and IgA-subclasses will be addressed. We will determine if the IL- 6R inducing factor is a novel cytokine, and if so, we will purify and clone this factor in order to evaluate its role in B cell responses. Finally, we plan to determine the relative contribution of antigen- specific T cells and derived cytokines for B cell responses, with emphasis on fimbriae and hsps. this comprehensive analysis of Th cells and cytokines will provide the cellular and molecular basis for B cell responses in human PD and may be of predictive importance in classification of the different forms of this disease.

我们过去在这项资助上的工作直接涉及免疫方面， 牙周病（PD）的发展方法的分离， 牙龈单核细胞（GMC）从疾病部位。 我们已经表明 大量的免疫球蛋白（IG）分泌，斑点形成细胞 (SFC)主要局限于IgG亚类 (IgG1> IgG 2> IgG 3 = IgG 4），而对IgA亚类（IgA 1> IgA 2）应答的影响较小。 这个独特的系统使我们能够确定抗原的程度- 特异性SFC反应，我们发现牙龈卟啉单胞菌 菌毛和LPS是这些IgG和伊加的主要和次要成分， 子类别的响应。 因此，我们的工作表明， 在PD中观察到的主要IgG和IgA浆细胞应答， 多克隆和抗原特异性组分。 这在逻辑上导致了 GMC上清液（sups）中存在的细胞因子分析， 解释了这些独特的B细胞反应。 我们的研究 现在表明IL-6是GMC中主要的终末分化因子 sups，连同IL-5。 对这项提案来说，重要的是存在 诱导IL-6受体（IL-6 R）的潜在新型细胞因子 在人淋巴细胞上的表达。 我们计划继续这些令人兴奋的 研究通过解决细胞因子存在的精确模式， GMC中单个细胞类型，包括B细胞和浆细胞 应答 我们将重点关注IL-2、IL-4、IL-5、IL-6和IFN-γ， 用PCR和原位杂交技术检测了这些细胞因子在T细胞和T细胞中的表达。 辅助（Th）细胞亚群、B细胞、单核细胞/巨噬细胞）和非免疫细胞 （成纤维细胞和上皮细胞）细胞，其来自患有 警局 我们的初步研究表明，牙龈Th细胞可能是 诱导，部分是由于牙龈卟啉单胞菌菌毛的存在， 超抗原和热休克蛋白（HSPs）。 我们计划评估CD 4 + 从GMC的Th细胞对这些抗原的反应，并确定是否 hsps诱导Th 1型（IFN-γ和IL-2）和菌毛（超抗原） 主要激发Th 2（IL-4、IL-5和IL-6）应答。 由特定 克隆的CD 4 + Th细胞的衍生，我们可以确定相对的 Th 1和Th 2细胞对炎症和B细胞应答的贡献， 分别 转化生长因子β和白细胞介素4在转基因中的潜在作用 将讨论IgG和IgA亚类。 我们将决定是否- 6 R诱导因子是一种新的细胞因子，如果是这样，我们将纯化， 克隆该因子以评估其在B细胞应答中的作用。 最后，我们计划确定抗原的相对贡献- 用于B细胞应答的特异性T细胞和衍生的细胞因子， 强调菌毛和热休克蛋白。 Th细胞的综合分析 而细胞因子则为B细胞提供了细胞和分子基础 在人类PD中的反应，可能具有预测的重要性， 这种疾病的不同形式的分类。