| Summary: | 碩士 === 慈濟大學 === 分子生物及細胞生物研究所 === 93 === Systemic lupus erythematosus (SLE) is a chronic inflammatory autoimmune disease. The exact cause of lupus remains a mystery, but it causes variable inflammatory destruction of skin、brain、heart、lungs、joints、blood-forming organs、kidneys and nervous system. The prevalence of SLE in Taiwan had been estimated as approximately fifty thousand (5/1000) predominantly female (male/female = 1/9) affected. In clinical diagnose, SLE patients after develop different kinds of autoantibodies, such as anti-double strand DNA (anti-dsDNA Ab)、anti-Smith antibody (anti-Sm Ab)、anti-small nuclear ribonucleoprotein antibody (anti-snRNP) and ect. Current investigations agreed that the mechanisms involved in promoting SLE including genetics (such as human leukocyte antigen (HLA) types)、complement deficiency、MHC、sex hormone、environmental factor (such as drug)、virus infection、ultraviolet light and stress.
Previous studies suggested that virus infection including Cytomegalovirus infection could increase the risk in the development of SLE. Cytomegalovirus infection is known to induce autoimmune diseases in mice. Our study focused on pp65 lower matrix phosphoprotein of HCMV (Human cytomegalovirus). The pp65 antigen has been reported to stimulate cell-mediated immunity by human and animal subjects following infection or immunization. However, the B cell epitope for pp65 remain elusive due to normal blood donors seldom develop antibody to it. We have revealed that adult SLE patients have elevated anti-pp65 activity. In order to verify the possible autoimmune associated B cell epitope, the pp65 antigen was divided into 501、510 and 672 nucleotide fragments, which expend the entire length of pp65 gene. Subsequently, these fragments were cloned into pGEX-KG and pET30 forming pp65-1、pp65-2、pp65-3 and pp65-F (full length) respectively. Expressed full-length pp65 and its fragments are examined against sera from SLE, none-SLE connective tissue disease patients (CTD), and normal control via western blotting. According to western blotting analysis the pp65-3 was recognized by SLE sera at a such higher frequency than pp65-1、2. Therefore, this study suggested that pp65-3 fragment contains B cell epitopes that could play a role in promoting SLE development.
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