Mechanisms of Immune Modulation and Periodontal Disease

免疫调节与牙周病的机制

• 批准号: 6606156
• 负责人: Jannet Katz
• 金额: $ 29.06万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6606156
• 关键词: B cell lymphoma; Bacteroides gingivalis; CD40 molecule; SDS polyacrylamide gel electrophoresis; bacteria infection mechanism; bacterial antigens; bacterial genetics; cellular immunity; cholera toxin; cytokine; enzyme linked immunosorbent assay; flow cytometry; gene expression; immunization; immunogenetics; immunoglobulin G; immunomodulators; immunoregulation; laboratory mouse; monoclonal antibody; mucosal immunity; pathologic process; periodontium disorder; phospholipids; surface antigens; western blottings

Periodontal disease is the result of the host response to periodontal pathogens such as Porphyromonas gingivalis. P. gingivalis is a black-pigmented, Gram-negative pathogen, which expresses various virulence factors implicated in disease. The overall goal of this project is to define the cellular mechanism(s) involved in the induction of host responses protective against P. gingivalis infection. The studies proposed will concentrate on delineating the mechanisms by which adjuvants modulate host responses to P. gingivalis antigens. Emphasis will be placed on the role of B7 co-stimulatory molecules in host responses and in infection by P. gingivalis, and on the involvement of cytokines, specific target cells, and other cell surface receptors. Specifically, there are plans to: 1) Determine the immunogenicity of the recombinant catalytic domain of the lysine-specific protease Kgp (rKgp-CAT) and the HagA repeat domain (rHArep) of P. gingivalis, and the effect of the B subunit of cholera toxin (CTB) and monophosphoryl lipid A (MPL) in modulating responses to these antigens following intranasal immunization, The level, isotype and IgG subclass of antibodies induced to HArep or Kgp-CAT in serum and external secretion$ will be measured by ELISA. The effects of CTB and MPL in enhancing/shifting responses will be further characterized by measuring antigen-specific proliferation and cytokine production. The effect of the responses, especially the nature of the antibodies, on protection will also be determined. 2) Determine the mechanisms by which the adjuvants CTB and MPL modulate the induction of the immune response. These studies will determine the involvement of the co-stimulatory B7 molecules in the immunoenhancing ability of the adjuvants and in P. gingivalis infection, the target cells affected by the adjuvants and the association between MPL, the Toll-like receptors and B7 co-stimulatory molecules. Groups of mice deficient in B7-l (B7-1-/-), B7-2 (B7-2j or both (B7-1/B7-2-/-) molecules and normal controls will be immunized with HArep or Kgp-CAT with and without CTB or MIPL. The levels of antibody and antibody-secreting cells will be assessed by ELISA and ELISPOT method. The effect of adjuvants on the expression of B7 molecules and of CD4OL will be analyzed by FACS. Differential regulation of cytokines induced and proliferative responses will also be assessed. The involvement of B7 co-stimulation in P. gingivalis infection will be assessed in B7 deficient mice. These studies will provide information on the mechanisms by which mucosal adjuvants modulate host immune responses, the target cells through which their adjuvanticity is exerted and the involvement of co-stimulatory signals provided by B7 molecules and of Toll-like receptors. An understanding of these processes is crucial for the development of means to interrupt/prevent periodontal disease pathogenesis by Porphyromonas gingivalis. Knowledge on the host effects induced by a potential periodontal vaccine will lead to the development of the therapeutic manipulation of effector function leading to the amelioration or prevention of periodontal disease and associated systemic diseases.

牙周病是宿主反应的结果 牙周病原体，例如牙龈卟啉单胞菌。牙龈卟啉单胞菌是 黑色素革兰氏阴性病原体，表达多种毒力 与疾病有关的因素。该项目的总体目标是定义 诱导宿主反应的细胞机制 预防牙龈卟啉单胞菌感染。拟议的研究将 专注于描绘佐剂调节宿主的机制 对牙龈卟啉单胞菌抗原的反应。重点将放在B7的作用上 宿主反应和牙龈卟啉单胞菌感染中的共刺激分子， 以及细胞因子、特定靶细胞和其他细胞的参与 表面受体。具体来说，计划： 1) 确定 赖氨酸特异性重组催化结构域的免疫原性 牙龈卟啉单胞菌的蛋白酶 Kgp (rKgp-CAT) 和 HagA 重复结构域 (rHArep)， 以及霍乱毒素 (CTB) B 亚基和单磷酰脂质的作用 A (MPL) 在鼻内调节对这些抗原的反应 免疫接种，诱导产生的抗体的水平、同种型和 IgG 亚类 血清和外部分泌物中的HArep或Kgp-CAT将通过ELISA测量。 CTB 和 MPL 在增强/改变反应方面的作用将进一步增强 通过测量抗原特异性增殖和细胞因子来表征 生产。反应的效果，尤其是反应的性质 抗体，保护作用也将被确定。 2）确定机制 佐剂CTB和MPL通过其调节免疫的诱导 回复。这些研究将确定共刺激 B7 的参与 佐剂和牙龈卟啉单胞菌免疫增强能力中的分子 感染、受佐剂影响的靶细胞及其关联 MPL、Toll 样受体和 B7 共刺激分子之间。团体 缺乏 B7-1 (B7-1-/-)、B7-2 (B7-2j 或两者 (B7-1/B7-2-/-) 分子的小鼠 正常对照将用 HArep 或 Kgp-CAT 进行免疫接种，有或无 CTB 或 MIPL。抗体和抗体分泌细胞的水平将是 通过ELISA和ELISPOT方法进行评估。佐剂对表达的影响 B7分子和CD4OL的分析将通过FACS进行分析。差别化监管 还将评估细胞因子诱导的和增殖反应。这 B7 共刺激参与牙龈卟啉单胞菌感染的评估将在 B7 缺陷小鼠。这些研究将提供有关机制的信息 哪些粘膜佐剂调节宿主免疫反应，靶细胞 通过它们发挥辅助作用并参与 B7 分子和 Toll 样受体提供的共刺激信号。一个 了解这些过程对于开发方法至关重要 中断/预防牙龈卟啉单胞菌引起的牙周病发病机制。 关于潜在牙周疫苗引起的宿主效应的知识将 导致效应器功能治疗操作的发展 导致牙周病和相关疾病的改善或预防 全身性疾病。