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: 6104728
• 负责人: HIROSHI KIYONO
• 金额: $ 7.42万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-12-01 至 2000-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6104728
• 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.

我们过去在这笔赠款上的工作直接解决了免疫方面的问题 牙周病(PD)分离方法的发展 病变部位的牙周单个核细胞(GMC)。我们已经展示了 大量免疫球蛋白(Ig)分泌，斑点形成细胞 (SFC)，主要限于免疫球蛋白亚类 (IgG1和GT；IgG2和GT；IgG3=IgG4)，而对IgA亚类(IgA1和GT；IgA2)反应的影响较小。 这个独特的系统使我们能够确定抗原的程度- 帕金森病的特异性SFC反应，我们发现牙龈卟啉单胞菌 菌毛和脂多糖是这些免疫球蛋白和免疫球蛋白A的主要和次要成分。 分别为子类响应。因此，我们的工作表明， 帕金森病患者主要的免疫球蛋白和免疫球蛋白A浆细胞反应既有 多克隆和抗原特异性成分。这在逻辑上导致了 分析GMC上清液(SUPS)中存在的细胞因子，这将 解释了B细胞反应的这些独特的特征。我们的研究已经 目前研究表明IL-6是GMC的主要终末分化因子 SUPS与IL-5联合应用。对这项提议来说，重要的是 一种潜在的诱导IL-6受体(IL-6R)的新细胞因子 在人淋巴细胞上的表达。我们计划继续这些激动人心的活动 通过处理细胞因子的精确模式进行的研究 GMC中的单个细胞类型，包括B细胞和浆细胞 回应。我们将重点介绍IL-2、IL-4、IL-5、IL-6和干扰素-γ及其用途 这两种细胞因子在免疫(T)中的PCR和原位杂交检测 辅助性(Th)细胞亚群、B细胞、单核/巨噬细胞)和非免疫 (成纤维细胞和上皮细胞) 警察。我们的初步研究表明，牙龈Th细胞可能是 部分是由于牙龈假单胞菌菌毛的存在而引起的 超抗原和热休克蛋白(HSPs)。我们计划评估CD4+ 来自GMC的TH细胞对这些抗原的反应并确定 热休克蛋白可诱导Th1型(干扰素-γ和IL-2)和菌毛(超抗原) 主要诱导Th2(IL-4、IL-5和IL-6)反应。按特定 克隆的CD4+Th细胞的衍生，我们可以确定相对 Th1和Th2细胞在炎症和B细胞反应中的作用 分别进行了分析。转化生长因子-β和白介素4在转归中的潜在作用 将解决免疫球蛋白亚类和免疫球蛋白A亚类。我们会确定IL- 6R诱导因子是一种新的细胞因子，如果是这样的话，我们将纯化和 克隆该因子，以评价其在B细胞反应中的作用。 最后，我们计划确定抗原的相对贡献- B细胞反应的特异性T细胞和衍生细胞因子， 重点是菌毛和热休克蛋白。Th细胞的综合分析 细胞因子将为B细胞提供细胞和分子基础 在人类帕金森病中的反应，并可能在预测重要性 这种疾病的不同形式的分类。