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Hepatoma AFP (alpha-fetoprotein) specific artificial antigen-presenting cell inducing reagent kit

An artificial antigen and cell induction technology, applied in the field of cell biology, can solve the problems of expensive, difficult to expand DC cells in vitro, and low CTL killing activity

Inactive Publication Date: 2019-06-21
YINFENG JILIN BIOLOGICAL ENG TECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The most commonly used method is to use autologous dendritic cells as antigen-presenting cells to obtain specific CTL cells. This method requires patients to provide a large number of white blood cells, and it is very expensive to culture induced DC cells in the laboratory. Expanding DC cells in vitro Currently a technical problem
More critically, the specificity of CTL cells induced by DCs in vitro is low, and the specific killing activity of CTLs is low

Method used

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  • Hepatoma AFP (alpha-fetoprotein) specific artificial antigen-presenting cell inducing reagent kit
  • Hepatoma AFP (alpha-fetoprotein) specific artificial antigen-presenting cell inducing reagent kit
  • Hepatoma AFP (alpha-fetoprotein) specific artificial antigen-presenting cell inducing reagent kit

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Embodiment Construction

[0024] The liver cancer AFP-specific artificial antigen-presenting cell induction kit provided by the invention comprises the following components:

[0025] HLA-A2-Ig dimer, purchased from BD Company, stored at 4°C;

[0026] CD28mAb antibody, purchased from BD Company, stored at 4°C;

[0027] Magnetic beads, preferably BeaverBeads Mag MHS magnetic beads, stored at 4°C;

[0028] AFP polypeptide, the amino acid sequence of which is shown in SEQ ID NO: 1, stored at -80°C;

[0029] Magnetic bead resuspension buffer, preferably borate buffer, its preparation method is: dissolve sodium borate in ultrapure water, configure it into a 0.1M solution, adjust the pH to 7.0, and store at 4°C;

[0030] Magnetic bead washing buffer, its preparation method is: PBS 956ml without calcium and magnesium ions, add 30ml of human AB serum, add EDTA to a final concentration of 2mM, add sodium azide 0.1g / L, sterilize with a 0.22um filter, Store at 4°C.

[0031] SEQ ID NO: 1

[0032] MKWVESIFLIFLL...

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Abstract

The invention discloses a hepatoma AFP (alpha-fetoprotein) specific artificial antigen-presenting cell inducing reagent kit, belongs to the technical field of cytobiology and solves the problems of the prior art that in-vitro amplification of DC cells is difficult and acquisition of specific CTLs (cytotoxic T Lymphocytes) by in-vitro stimulation of CD8+T cells is low in efficiency. The kit hereincomprises an HLA-A2-Ig dimer, an anti-CD28mAb antibody, magnetic beads, and AFP polypeptides. The kit herein is effective in amplifying antigen-specific CTLs in vitro, with specificity maintained formonths; CD*+T cells are stimulated for hepatoma-specific aAPC, the CTLs account for 95% and above the total cell quantity after one week of cultivation, and the amplification times is 1000 and above.The cell quantity can be further increased by repeatedly stimulating the AFP specific CTLs via aAPC. The attained CTLs present effect memory cell features in appearance, and can secrete TNF alpha andIFN gamma.

Description

technical field [0001] The invention relates to the technical field of cell biology, in particular to a liver cancer AFP-specific artificial antigen-presenting cell induction kit. Background technique [0002] Antigen-specific T cells (AST) are the core cells that mediate the adaptive immune response, and their quantity and function directly reflect the functional status of heterogeneous cellular immunity in the human body. Detecting the number and reactivity of antigen-specific T cells is of great significance in the study of pathogenesis and individualized clinical treatment of diseases such as infection, tumor, autoimmune disease, and transplant rejection. [0003] The activation of the body's antigen-specific immune function requires antigen-presenting cells (antigen presenting cells, APCs) to transmit specific pathogenic signals to T cells to generate antigen-specific T lymphocytes. There are three types of antigen-presenting cells in the human body, including dendriti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783
Inventor 魏本杰李博徐峰波高佳左红斌
Owner YINFENG JILIN BIOLOGICAL ENG TECH CO LTD
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