人BAFF基因轉染細胞及其單克隆抗體的生物學特性
[Abstract]:BAFF (B cell-activating factor belong to the TNF family) is a new molecule found in 1999, mainly expressed in monocytes, macrophages, dendritic cells, activated T lymphocytes and neutrophils. Its receptors include the specific binding receptors BAFF-RnTACI and BCMA.The three receptors are mainly expressed in T and B lymphocytes. BAFF and its receptors are important members of the TNF/TNFR superfamily. It has been found that BAFF signal plays an important role in the differentiation, development and antibody type conversion of B lymphocytes, as well as the maintenance of germinal center, and has co-stimulatory effect on T lymphocytes, and is associated with many autoimmune diseases such as SLERA and SS. Tumors such as chronic B lymphocytic leukemia B cell non Hodgkin's disease and multiple myeloma are associated with allograft rejection of heart and other organs. However, the basic research of BAFF needs to be further studied. In order to study the BAFF signal, we first cloned the human full-length BAFF molecule, established the BAFF gene transfection cells, and used it as the immunogen, successfully developed a monoclonal antibody against human BAFF, and studied its biological characteristics. The expression and significance of BAFF on T lymphocytes from peripheral blood and pleural effusion were analyzed. At the same time, the full-length human TACI gene of the receptor was cloned, and the gene transfected cells expressing TACI-Fc fusion protein were established, and the biological effects of the TACI-Fc fusion protein were discussed. 1. Construction of human BAFF gene transfected cells and effect of BAFF molecule on B cell function. 1. Cloning of Human BAFF Gene and Construction of Retrovirus expression Vector according to the sequence of BAFF Gene (OeneBank:NM 006573), The full-length BAFF gene was cloned from the peripheral blood mononuclear cells of healthy people by RT-PCR method. The gene was inserted into the pMD18-T vector and transformed into the competent E. coli Top10. The recombinant plasmid T-BAFF was sequenced correctly. Using PCR method, the full-length BAFF gene was amplified from the recombinant plasmid T-BAFF, and the restriction sites EcoR I and BamH I were introduced into the retroviral expression vector pSIV1. The recombinant expression vector pSIV1-BAFF was sequenced correctly. 2. BAFF gene transfected pSIV1-BAFF cells were constructed by liposome transfection method. After pHIT456 and pHIT60 were mixed, the packaging cells 293T cells were transfected with liposome transfection method. An infectious recombinant retrovirus was obtained. Will contain full BAFF recombination reversal
【學位授予單位】：蘇州大學
【學位級別】：博士
【學位授予年份】：2005
【分類號】：R392
【共引文獻】
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