T cell receptor and application thereof

A cell receptor and cell technology, applied in the fields of genetic engineering and cellular immunology, can solve the problems of poor treatment of solid tumors, difficult control of cytokine storm, limited selection of CAR-T targets, etc.

Pending Publication Date: 2021-12-10
深圳大学总医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the limitations of current CAR-T therapy still exist obviously. First of all, CAR-T can choose limited targets, and the treatment of solid tumors has not yet had a good effect.
Moreover, adverse reactions brought about by CAR-T therapy, such as cytokine storm, are sometimes difficult to control

Method used

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  • T cell receptor and application thereof
  • T cell receptor and application thereof
  • T cell receptor and application thereof

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment approach

[0153] The present invention will be described in further detail below in conjunction with the accompanying drawings and embodiments.

[0154] The following examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

Embodiment 1

[0155] Example 1 High-throughput sequencing screening of TCR sequences

[0156] 1. RNA extraction and quality control

[0157] Peripheral blood samples were collected from patients with acute myeloid leukemia who were free of hepatitis A, hepatitis B, hepatitis C, hepatitis E, AIDS, syphilis, gonorrhea and tuberculosis infection.

[0158]Slowly add peripheral blood into 5mL Ficoll and centrifuge at 2000rpm for 15min. Aspirate the middle buffy coat, add 0.9% normal saline, count the number of mononuclear cells, centrifuge at 1000 rpm for 5 min, and sort T cells with magnetic beads. RNA was extracted from the treated cell samples by the TRIzol method, and the quality of the extracted RNA was tested using Qubit RNA HS Assay kits, and the integrity of the RNA was detected by a 2100 bioanalyzer (Agilent).

[0159] 2. Reverse transcription and library preparation

[0160] Use Repertoire Analysis Kit (Human TCR alpha, beta) to build the library. Thaw all reagents on ice, mix and ...

Embodiment 2

[0213] Example 2 Construction of TCR-T

[0214] 1. Using the results of high-throughput sequencing and data analysis, fully synthesize the relevant sequences, and connect them to the pCDH vector through the XbaI / SalI restriction endonuclease cut point.

[0215] 2. Preparation of TCR-T cells

[0216]Collect peripheral blood from healthy people, slowly add peripheral blood to Ficoll, and centrifuge at 2000rpm for 15min. Aspirate the middle buffy coat, add 0.9% normal saline, count the number of mononuclear cells, and centrifuge at 1000rpm for 5min. Magnetic bead sorting of T cells. At the same time, CD3 / CD28 antibody-coupled magnetic beads were added for stimulation. After 24 hours and 48 hours, virus infection was added twice. IL-2 was added during virus infection, and TCR-T cells were obtained by culturing for 3-20 days.

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Abstract

The invention discloses a T cell receptor and application thereof, and also discloses related polypeptides and proteins, as well as related nucleic acids, vectors, cells, compositions and chimeric molecules.

Description

technical field [0001] The invention belongs to the fields of cellular immunology and genetic engineering, and relates to T cell receptors and applications thereof. Background technique [0002] As an indispensable large class of immune cells in the human immune system, T lymphocytes play a vital role in protecting the body from foreign pathogenic microorganisms and maintaining the balance of the body's immune system. T lymphocytes are derived from bone marrow, and under the induction of thymus hormones, mature in the thymus microenvironment to form highly differentiated immune cells with immune activity. Mature T lymphocytes enter the peripheral immune organs of the human body along with the blood flow, and can recirculate through lymphatic vessels and peripheral blood to exert cellular immunity and immune regulation functions. [0003] At present, in addition to traditional surgery, chemotherapy, and radiotherapy, immunotherapy has rapidly developed into a recognized and ...

Claims

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Application Information

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IPC IPC(8): C07K14/725C12N15/12C12N15/867C12N5/10A61K39/00A61K35/17A61P37/04A61P35/00A61P35/02
CPCC07K14/7051C12N15/86C12N5/0636A61K39/001102A61K35/17A61P37/04A61P35/00A61P35/02C12N2740/15043C12N2740/10043C12N2510/00
Inventor 王智鼎于力
Owner 深圳大学总医院
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