MYCOPLASMAS AND CHLAMYDIAE AND RHEUMATOID ARTHRITIS

支原体、衣原体和类风湿性关节炎

• 批准号: 2006343
• 负责人: GAIL H. CASSELL
• 金额: $ 27.36万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-01-15 至 1998-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2006343
• 关键词: Chlamydiaceae; Mycoplasma; bacterial cytopathogenic effect; enzyme linked immunosorbent assay; genotype; histocompatibility typing; human subject; immunoelectron microscopy; in situ hybridization; microorganism classification; microorganism culture; monoclonal antibody; polymerase chain reaction; rheumatoid arthritis; synovial fluid; tissue /cell preparation; western blottings

It has been increasingly suggested that some cases of rheumatoid arthritis (RA) and allied diseases may arise from the development of immunopathological responses following established or latent infection with a possible wide range of microorganisms. An extension of this view is that the different clinical syndromes within this group of disorders arise from immunoregulatory defects, the expression of which could be influenced by both genetic and environmental factors. Recent work in experimental animals regarding susceptibility to infectious agents strongly suggest that this is entirely possible. Furthermore, there are known rheumatic diseases of man for which there is now cogent evidence that infection and genetic influence play a role. Despite many attempts to identify the inciting infectious agent(s)in RA, results to date seem to be a negative catalogue of technical artifact and interpretative error. Careful examination of experimental protocols used in these studies coupled with recent conceptual and technical advances in microbiology, immunology and molecular biology provide plausible explanations of past failure. All indications are that the time has come to initiate another search. In general, previous investigations of RA have suffered from ill-defined parameters for disease classification and lack of interdisciplinary cooperation. The early stages of synovitis, which may provide valuable clues to pathogenesis, have received little study. The purpose of the present proposal is to define those infectious agents associated with rheumatoid arthritis of recent onset (2 to 12 mos.) in well characterized patient populations. The ability to detect organisms within affected joints will be enhanced by evaluation of synovial fluid and by evaluation of synovial tissue collected by arthroscopy. Current evidence suggests that two of the most likely agents to be involved in chronic arthritis in humans are mycoplasmas and chlamydiae. Thus, the present study will focus on these two organisms. Improved methods will be used for isolation of mycoplasmas and chlamydiae from synovial fluid and tissue, peripheral blood monocytes, and when indicated mucosal surfaces. Isolates will be identified using polyclonal and monoclonal antibodies. In addition, presence of microorganisms will be detected by amplication of nucleic acid by the highly sensitive and specific polymerase chain reaction (PCR). PCR will also be used to confirm species identity of mycoplasmal and chlamydial isolates. Detection of microorganisms directly in affected joints will be verified using immunoelectron microscopy and in situ hybridization. Detection of specific antibody to any organism detected in affected joints will be determined using enzyme-linked immunosorbent assays in conjunction with immunoblotting. Patient MHC genotypes will also be determined. Demographic and experimental data will be collected in such a way that all parameters may be correlated one with the other. All data obtained in RA will be compared to matched controls with osteoarthritis. We propose to accomplish these objectives by combining the investigative talents of individuals with backgrounds in clinical rheumatology, microbiology, molecular biology, immunology, genetics, epidemiology, and computer technology.

越来越多的人认为某些类风湿性关节炎病例 （RA）和相关疾病可能是由于以下疾病的发展而引起的： 已确诊或潜伏感染后的免疫病理反应 可能存在多种微生物。这种观点的延伸是 这组疾病中会出现不同的临床综合征 免疫调节缺陷，其表达可能受到影响 由遗传和环境因素共同影响。最近的实验工作 动物对传染性病原体的易感性强烈建议 这完全有可能。此外，已知还有风湿病 现在有确凿的证据表明感染和 遗传影响发挥作用。尽管进行了多次尝试来识别 在 RA 中煽动传染源，迄今为止的结果似乎呈阴性 技术工件和解释错误的目录。小心 检查这些研究中使用的实验方案以及 微生物学、免疫学和微生物学的最新概念和技术进展 分子生物学为过去的失败提供了合理的解释。全部 有迹象表明，现在是启动另一次搜索的时候了。在 一般而言，以前对 RA 的研究都存在定义不明确的问题 疾病分类参数和缺乏跨学科 合作。滑膜炎的早期阶段，这可能提供有价值的信息 发病机制的线索，很少有研究。目的 目前的提案是定义那些与 近期发病（2 至 12 个月）的类风湿性关节炎特征明确 患者人群。检测受影响的生物体的能力 通过评估滑液和评估将增强关节 通过关节镜检查收集的滑膜组织。目前的证据表明 两种最有可能与慢性关节炎有关的药物 人类有支原体和衣原体。因此，本研究将重点 在这两种生物上。将使用改进的方法来隔离 来自外周滑液和组织的支原体和衣原体 血液单核细胞，以及粘膜表面（当有指示时）。分离株将是 使用多克隆和单克隆抗体进行鉴定。此外， 通过核酸扩增来检测微生物的存在 通过高度灵敏和特异性的聚合酶链式反应 (PCR)。聚合酶链式反应 也将用于确认支原体的物种身份 衣原体分离株。直接检测受影响的微生物 将使用免疫电子显微镜和原位验证接头 杂交。检测任何生物体的特异性抗体 将使用酶联免疫吸附剂确定受影响的关节 与免疫印迹结合进行测定。患者 MHC 基因型也会 被确定。人口统计和实验数据将在此类中收集 所有参数都可以相互关联的一种方式。所有数据 在 RA 中获得的结果将与患有骨关节炎的匹配对照进行比较。 我们建议通过结合调查来实现这些目标 具有临床风湿病学背景的人才， 微生物学、分子生物学、免疫学、遗传学、流行病学和 计算机技术。