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MR gene order of diffuse large B cell lymphoma related antigen specific TCR V alpha 6 subfamily and uses thereof

A gene sequence, lymphoma technology, applied in the direction of receptor/cell surface antigen/cell surface determinant, application, gene therapy, etc., can solve the problem of understanding antigen

Inactive Publication Date: 2008-07-23
JINAN UNIVERSITY +1
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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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  • MR gene order of diffuse large B cell lymphoma related antigen specific TCR V alpha 6 subfamily and uses thereof

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Embodiment

[0034] (1) Analysis of antigen-specific TCR gene sequence

[0035]The method for analyzing the length and base sequence of CDR3 of TCR Vα gene profile is to analyze the length of CDR3 of 29 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 subfamily genes in patients . As shown in Figure ...

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Abstract

The invention discloses a nucleotide sequence and the application of an antigen-specific T cell receptor of the diffusive large B cell lymphoma. According to the RT-PCR genescan analysis, the TCR V Alpha 6 subfamilies in the peripheral blood of patients shows an obviously monoclonal state, and the V-N-J region, namely, the complementary determining region 3, where the PCR products conduct the sequence analysis of the specific TCR sequence, is a part recognizably bound to the antigenic specificity; the genescan analysis also shows that the TCR of the clonal T cell is a specific TCR produced due to the stimulus of the DLBL associated antigens, and is the basis and key for performing DLBL molecular targeted adoptive immunotherapy by means of the TCR transgenic technique. The nucleotide sequence of the specific TCR provides a significant basis for further carrying out lymphoma-specific cellular immunotherapy.

Description

technical field [0001] The invention relates to a T cell receptor (TCR) nucleotide sequence, in particular to the MR gene sequence and application of diffuse large B cell lymphoma (DLBL) related antigen-specific TCR Vα6 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 ne...

Claims

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Application Information

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IPC IPC(8): C12N15/12C12N15/10C07K14/725A61K31/7088A61K48/00A61P37/02
Inventor 李扬秋谭获尹青松杨力建陈少华叶静梅吴秀丽
Owner JINAN UNIVERSITY
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