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解决了免疫方面的问题 通过开发分离牙周病（PD）的方法 来自疾病部位的牙龈单核细胞（GMC）。 我们已经展示了 大量分泌免疫球蛋白 (Ig) 的斑点形成细胞 (SFC) 发生，主要限于 IgG 亚类 (IgG1>IgG2>IgG3=IgG4)，而 IgA 亚类 (IgA1>IgA2) 反应则较少。 这种独特的系统使我们能够确定抗原的程度 PD 中特定的 SFC 反应，我们发现牙龈卟啉单胞菌 菌毛和 LPS 是这些 IgG 和 IgA 的主要和次要成分 分别是子类响应。 因此，我们的工作表明 PD 中主要的 IgG 和 IgA 浆细胞反应同时具有 多克隆和抗原特异性成分。 这从逻辑上导致了 对 GMC 上清液 (sups) 中存在的细胞因子进行分析，这将 解释了 B 细胞反应的这些独特特征。 我们的研究有 现在表明 IL-6 是 GMC 中主要的终末分化因子 与 IL-5 一起。 该提案的重要意义在于存在 诱导 IL-6 受体 (IL-6R) 的潜在新型细胞因子 人淋巴细胞上的表达。 我们计划继续这些令人兴奋的活动 通过解决存在于细胞因子的精确模式的研究 GMC 中的单个细胞类型，包括 B 细胞和浆细胞 回应。 我们将重点关注 IL-2、IL-4、IL-5、IL-6 和 IFN-gamma，并使用 PCR 和原位杂交对这些细胞因子在免疫（T 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）来自患者组织的细胞 PD。 我们的初步研究表明牙龈 Th 细胞可能 部分是由牙龈卟啉单胞菌菌毛的存在引起的 超抗原和热休克蛋白（hsps）。 我们计划评估 CD4+ 来自 GMC 的 Th 细胞对这些抗原作出反应并确定是否 hsps 诱导 Th1 型（IFN-γ 和 IL-2）和菌毛（超抗原） 主要引起 Th2（IL-4、IL-5 和 IL-6）反应。 按具体 克隆 CD4+ Th 细胞的衍生，我们可以确定相对 Th1 和 Th2 细胞对炎症和 B 细胞反应的贡献， 分别。 TGF-β 和 IL-4 在转换中的潜在作用 将讨论 IgG 和 IgA 亚类。 我们将确定 IL- 6R诱导因子是一种新型细胞因子，如果是的话，我们将纯化并纯化 克隆该因子以评估其在 B 细胞反应中的作用。 最后，我们计划确定抗原的相对贡献 用于 B 细胞反应的特定 T 细胞和衍生细胞因子， 强调菌毛和热休克蛋白。 对 Th 细胞的全面分析 细胞因子将为B细胞提供细胞和分子基础 人类帕金森病的反应，可能具有预测重要性 该疾病的不同形式的分类。