Analysis Of The TCR CDR3 Spectratyping And Monitor Of The TCR Gene Rearrangement In The Process Of TCRαβT Cells Response Or Tolerance

Objective(1) Establishing the immunoscope spectratyping technique for monitoring CDR3 polymorphism and length distribution of human TCR αβ T cells. Analysis of the CDR3 size distribution of TCR up T cells in the Peripheral Blood of healthy controls, patients with Chronic Hepatitis B (CHB), patients with T-lineage acute lymphoblastic leukemia (T-ALL), and patients with β-Mediterranean anaemia (the PBMC samples of pre-transplanted and post-transplanted with allogeneic hematopoietic stem cell). For the purpose to study of the mechanisms of T cells immune response or tolerance, and individualized immunotherapy of the infections disease, tumor, auto-immune disease. At the same time for the purpose to monitor the shift of human TCR repertoires in the research of TCR second rearrangement.(2) Establishing a special ligation-mediated PCR for analyzing the recombination signal sequence(RSS) broken ends of TCR BD gene. And identified mRNA expression of recombination activating gene (RAG) , terminal deoxynucleotidyl transferase (TdT), Ku70 & Ku80 by reverse transcriptase polymerase chain reaction; Primary study of the TCR second rearrangement in the PBMC of patients with CHB, patients with T-ALL, samples of Molt-4/LoVo/U937 , It would be applied to TCR second rearrangement and T cells response and tolerance in the corresponding diseases of T cells response.(3) Identifying the Anti-human CD3×Anti-carcinoma embryo antigen (CEA) bi-specific antibody (BsAb) by western blotting ; Study of the Anti-CD3×Anti-CEA BsAb mediated the conjugation between T cells (CD3 positive) and target cell (CEA positive); Analysis of the mechanisms for peripheral blood lymphocyte (PBL) killing the tumor cells mediated by Anti-CD3...