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Y gene order of diffused large B cell lymphoid tumor related antigen specific TCR V Beta 13 subfamily

A technology of gene sequence and B cells, applied in the field of nucleotide sequence of T cell receptors, can solve problems such as antigen understanding

Inactive Publication Date: 2008-05-21
JINAN UNIVERSITY +1
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

At present, the research on lymphoma at home and abroad mainly focuses on T-cell lymphoma, while there are few research materials on B-lymphocyte lymphoma.

Method used

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  • Y gene order of diffused large B cell lymphoid tumor related antigen specific TCR V Beta 13 subfamily

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Experimental program
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Effect test

Embodiment

[0043] (1) Analysis of antigen-specific TCR gene sequence

[0044] There are many methods for analyzing the length and base sequence of CDR3 of TCR Vβ gene profile, and the common method is to use RT-PCR-gene scanning method to analyze the length of CDR3 of 24 subfamilies of TCR Vβ. First, the peripheral blood of patients with DLBL was collected, and the peripheral blood mononuclear cells were separated by Ficoll stratification method, and 1*10 7 RNA was extracted from PBMCs (peripheral blood mononuclear cells), and cDNA was synthesized by RT-PCR. At the same time, the gene sequence of the TCRβ chain was found in the NCBI gene bank, and the positions of the V and C regions of each subfamily in the genome were marked. According to the cDNA of 24 Vβ subfamily genes, corresponding 24 Vβ specific upstream primers were designed, and a Cβ downstream primer was set in the Cβ region, and the V-NDN- JC District. As shown in Figure 1, fluorescein Fam marks the Cβ region of the PCR pr...

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Abstract

The invention discloses a nucleic acid sequence of specific T cell receptor, which aims at the diffuse large B-cell lymphoma associated antigen. The RT-PCR gene scan analysis shows that the TCR V Beta 13 subfamily in peripheral blood of DLBL patients has obvious monoclone; the V-NDN-J region of specific TCR sequence is discovered by the sequence analysis of the PCR product, which is the complementarity determining region 3; the complementarity determining region 3 is the specific recognition and binding site of the antigen, which shows that the TCR of the clonal T cell is the specific TCR generated by the stimulation of DLBL associated antigen. The invention is the basis and key for the DLBL molecular target adoptive immunotherapy through TCR transgenic technology. The specific TCR nucleic acid sequence provides important discovery for the further development of specific cellular immunotherapy of the lymphoma.

Description

technical field [0001] The invention relates to a T cell receptor (TCR) nucleotide sequence, in particular to a diffuse large B cell lymphoma (DLBL) related antigen-specific TCR Vβ13 subfamily Y gene sequence. Background technique [0002] In recent years, the incidence of non-Hodgkin's lymphoma (NHL) has continued to rise. It is estimated that NHL will account for 4.4% of aggressive malignancies and 4.8% of all cancer deaths in the early 21st century. Among them, diffuse large B-cell lymphoma (DLBL) is the most common NHL, accounting for about 30% to 40%, and elderly patients over 60 years old account for 70%. For more than 20 years, CHOP regimen (cyclophosphamide, doxorubicin, vincristine, prednisone) was mainly used as the main chemotherapy. However, it has toxic effects on some elderly people, and the long-term survival rate is low. Although the recent emergence of targeted immunotherapy such as rituximab (CD20 monoclonal antibody) has achieved new curative effects, d...

Claims

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Application Information

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IPC IPC(8): C12N15/12C07K14/435
Inventor 李扬秋谭获尹青松陈少华杨力建叶静梅李萡
Owner JINAN UNIVERSITY
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