A tumor immune T cell detection kit and method

A detection kit and cell technology, applied in the field of precise biomedical detection, can solve the problems of imperfect detection targets, adverse cytokines, T cell inactivation, etc., and achieve the effects of low cost, high sensitivity, and accurate quantification.

Inactive Publication Date: 2017-08-29
成都克里斯博生物科技有限公司
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Problems solved by technology

However, the current immune cell quantitative PCR detection targets for tumor immunity are not perfect
In view of the lack of effective detection of immune cell receptors, ligands, cytokines and other gene expressions in the current tumor immunity: (1) Among various immune cells, peripheral blood T cells have strong cytotoxicity to tumor cells; (2) There are a variety of activating receptors involved in the killing effect of T cells on tumor cells, and different types of tumors respond to d...

Method used

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  • A tumor immune T cell detection kit and method
  • A tumor immune T cell detection kit and method
  • A tumor immune T cell detection kit and method

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Experimental program
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Effect test

Embodiment Construction

[0035] 1. Purification and culture of PBMC cells

[0036] Reagent:

[0037]

[0038] operate:

[0039] 1. Take 15ml of human peripheral blood, anticoagulate (EATA or heparin) treatment, room temperature for 15 minutes.

[0040] 2. Centrifuge at 350g for 5 minutes at room temperature, and draw out the plasma for later use. Add 1 volume of PBS buffer to the blood cells to dilute and mix. (If using blood components, add 1 volume of PBS buffer, dilute and mix.)

[0041] 3. Take a 15ml centrifuge tube and add 5ml of lymphocyte separation medium. Slowly add 5ml of diluted blood cells to the upper layer.

[0042] 4. Centrifuge at 450g for 20 minutes at room temperature. From top to bottom, blood cells are divided into platelet layer, white blood cell layer (buffy coat) and red blood cell layer.

[0043] 5. Aspirate the platelet layer (approximately 2ml) with a pipette. Transfer the buffy coat (buffy coat, about 3ml) to a new 15ml centrifuge tube.

[0044] 6. Add 10ml PBS ...

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Abstract

A tumor immune T cell detection kit and method are provided. The kit includes specific primers for detecting specific quantitative expression of immune-related target genes in human peripheral blood T cells through subjecting the target genes to PCR amplification based on SYBR-Green quantitative PCR amplification. The immune-related target genes include receptor and ligand related target genes, NK cell related target genes and key cytokine genes. Expression profiles of the immune system related target genes in the human peripheral blood T cells can be accurately and quantitatively detected by the kit and the method successfully so that targets can be provided for accurate immune treatment of tumor patients. In a treatment process, cytokine storm response to immune treatment of a patient can be detected timely, thus providing guidance for avoiding adverse reactions. States of the immune system in the treatment process and after treatment are assessed, thus providing supports for adjusting the amount of an antibody medicine used for treatment or the feedback number of modified T cells and even target adjustments. The kit and the method are comprehensive in detection, rapid, quantitative, accurate, high in sensitivity and low in cost.

Description

technical field [0001] The invention belongs to the field of precision biomedical detection, and more specifically relates to the detection of quantitative expression profiles and specificity of immune system specific genes in human peripheral blood T cells related to preclinical and clinical research of tumor immune precision medicine by quantitative PCR amplification. PCR primers. Background technique [0002] Tumor immunotherapy is the application of immunological principles and methods to improve the immunogenicity of tumor cells, enhance the body's immune response, and use in vitro expansion of immune cells or infusion of effector drugs into patients to cooperate with the body's immune system to inhibit tumor growth (Kershaw et al ., 2013; Barrett et al., 2014; Melero et al., 2015). In the 1950s, Burnet and Thomas put forward the theory of "immune surveillance", which believed that the mutated tumor cells that often appear in the body can be recognized and eliminated b...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6851C12Q1/6886C12Q2600/158C12Q2600/166C12Q2563/107C12Q2545/114
Inventor 唐珂胡边
Owner 成都克里斯博生物科技有限公司
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