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P gene order of diffused large B cell lymphoid tumor related antigen specific TCR V Alpha 23 subfamily

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

Inactive Publication Date: 2008-05-21
THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT) +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • P gene order of diffused large B cell lymphoid tumor related antigen specific TCR V Alpha 23 subfamily

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Experimental program
Comparison scheme
Effect test

Embodiment

[0041] (1) Analysis of antigen-specific TCR gene sequence

[0042] There are many methods for analyzing the length and base sequence of CDR3 in the TCR Vα gene profile, and the common method is to analyze the length of CDR3 of the 94 subfamilies of TCR Vα by using RT-PCR-gene scanning method. 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 TCRα 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. Design corresponding 29 Vα-specific upstream primers based on the cDNA of 29 Vα subfamily genes, and set a Cα downstream primer in the Cα region, and use PCR technology to amplify the V-N-JC region generated by the TCR rearrangement of the 29 Vα subfami...

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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 Alpha 23 subfamily in peripheral blood of DLBL patients has obvious monoclone; the V-N-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.

Description

technical field [0001] The invention relates to a T cell receptor (TCR) nucleotide sequence, in particular to the P gene sequence of the diffuse large B cell lymphoma (DLBL) related antigen-specific TCR Vα23 subfamily. 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 e...

Claims

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Application Information

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IPC IPC(8): C12N15/12C07K14/435
Inventor 谭获李扬秋尹青松陈少华杨力建叶静梅
Owner THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT)
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