VIRAL ETIOLOGY OF SYSTEMIC LUPUS ERYTHEMATOSUS

系统性红斑狼疮的病毒病因

• 批准号: 6080896
• 负责人: LINDA Ann SPATZ
• 金额: $ 14.51万
• 依托单位: CITY COLLEGE OF NEW YORK
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-30 至 2001-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6080896
• 关键词: B lymphocyte; DNA; DNA binding protein; Epstein Barr virus; antiantibody; disease /disorder etiology; gene expression; genetically modified animals; laboratory mouse; mutant; nucleic acid sequence; systemic lupus erythematosus; virus cytopathogenic effect; virus protein

Systemic Lupus Erythematosus (SLE) is a chronic autoimmune disease characterized by the production of anti-dsDNA antibodies. The etiology of SLE is unknown but it has been postulated that viruses may play a role. One virus that has frequently been implicated in SLE is Epstein-Barr virus (EBV), a B-cell trophic virus capable of establishing a latent infection. High titers of antibody to EBV are found in the sera of young SLE patients. DNA by itself is not immunogenic, but studies with the polyoma virus T antigen have shown that the binding of this viral DNA binding protein to host DNA can render it immunogenic, and trigger an anti-dsDNA response. The aim of this project is to determine whether an EBV, DNA binding protein such as EBNA-1 can elicit the production of anti-dsDNA antibodies after binding to host DNA. Since mice are not permissive for EBV infection, one way to express EBNA-1 will be to immunize mice intramuscularly with EBV based vectors that can express EBNA-1 in mouse cells. Sera from these mice will be assayed for the presence of antibodies to EBNA1. Mice with antibodies to EBNA-1 will be subsequently tested for antibodies to dsDNA. It is proposed that the binding of EBNA-1 to dsDNA is critical for anti-dsDNA antibody production. To demonstrate this, mutants will be constructed that lack the ability to bind DNA. Mice will be immunized with these mutants to determine if they loose their capacity to elicit anti-dsDNA antibodies. Mice transgenic for the EBNA-1 sequence will also be tested for the presence of anti-dsDNA antibodies. If transgenic mice are tolerant to EBNA-1 and EBNA-1-DNA complexes, then B-cells expressing the transgene will be transferred into wild type mice and sera from the recipients will be assayed for the production of anti-dsDNA antibodies.

系统性红斑狼疮（SLE）是一种慢性自身免疫性疾病，其特征在于产生抗dsDNA抗体。系统性红斑狼疮的病因尚不清楚，但已假定病毒可能起作用。 一种经常与SLE有关的病毒是EB病毒（EBV），一种能够建立潜伏感染的B细胞营养型病毒。 年轻SLE患者血清中存在高滴度的EBV抗体。 DNA本身不是免疫原性的，但多瘤病毒T抗原的研究表明，这种病毒DNA结合蛋白与宿主DNA的结合可以使其具有免疫原性，并引发抗dsDNA应答。 本项目的目的是确定EB病毒DNA结合蛋白（如EBNA-1）与宿主DNA结合后是否能引发抗dsDNA抗体的产生。由于小鼠不允许EBV感染，表达EBNA-1的一种方法是用可在小鼠细胞中表达EBNA-1的基于EBV的载体肌内免疫小鼠。 将测定来自这些小鼠的血清中是否存在EBNA 1抗体。 随后测试具有EBNA-1抗体的小鼠的dsDNA抗体。 提出EBNA-1与dsDNA的结合对于抗dsDNA抗体的产生是关键的。 为了证明这一点，将构建缺乏结合DNA能力的突变体。 将用这些突变体免疫小鼠以确定它们是否失去引发抗dsDNA抗体的能力。还将测试EBNA-1序列转基因小鼠是否存在抗dsDNA抗体。 如果转基因小鼠耐受EBNA-1和EBNA-1-DNA复合物，则将表达转基因的B细胞转移到野生型小鼠中，并测定来自受体的血清中抗dsDNA抗体的产生。