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Initiative immune cell activation and amplification system and application thereof

An immune cell and expansion system technology, applied in the field of initial immune cell activation and expansion system, can solve the problem that patients cannot prepare and obtain CAR-T cells in vitro, so as to ensure the preparation quantity and quality, avoid functional decline, avoid Insufficient number of effects

Active Publication Date: 2021-07-09
THE SECOND AFFILIATED HOSPITAL OF ANHUI MEDICAL UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Adoptive cell therapy is a brand-new "anti-tumor active drug therapy technology" created on the basis of traditional anti-cancer treatment methods, which has created a new starting point for overcoming cancer. The product has achieved encouraging results and has a good application prospect, but at the same time, some problems have also been found. One of the key problems is that some patients cannot obtain the minimum amount of CAR-T cells required for treatment in vitro, or a small number of patients can obtain enough CAR-T cells. The number of CAR-T cells showed immune resistance after reinfusion, and there was no response to treatment

Method used

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  • Initiative immune cell activation and amplification system and application thereof
  • Initiative immune cell activation and amplification system and application thereof
  • Initiative immune cell activation and amplification system and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] 1. Preparation of activated medium

[0079] Preparation method of activation medium: take 15mL PRIME-XV T Cell CDM medium, add CD3 antibody to 100ng / mL, add CD28 antibody to 100ng / mL, add IL-2 to 200IU / mL, add IL-7 to 10ng / mL , add IL-15 to 10 ng / mL, add TGF-βR antibody to 50 ng / mL.

[0080] 2. Isolation of PBMCs

[0081] 2.1 Clinical patient A, with informed consent, collected peripheral blood samples intravenously. Receive the blood sample, wipe the surface with a dust-free cloth soaked in 75% alcohol for disinfection, and put it into a biological safety cabinet.

[0082] 2.2 Take 15mL blood sample and transfer it to a clean and sterile 50mL centrifuge tube.

[0083] 2.3 Add normal saline (NS) for injection to a volume of 30 mL in a 50 mL centrifuge tube, and mix the blood cells evenly.

[0084] 2.4 Transfer 15mL GE Ficoll Lymphocyte Separation Solution to a clean and sterile 50mL centrifuge tube.

[0085] 2.5 After the Ficoll is equilibrated at room temperature,...

Embodiment 2

[0152] 1. Preparation of activated medium

[0153] Preparation method of activation medium: Take 15mL PRIME-XV T Cell CDM medium, add CD3 antibody to 150ng / mL, add CD28 antibody to 75ng / mL, add IL-2 to 250IU / mL, add IL-7 to 15ng / mL , add IL-15 to 15ng / mL, add TGF-βR antibody to 100ng / mL.

[0154] 2. Separation of PBMCs

[0155] 2.1 Clinical patient B, with informed consent, collected peripheral blood samples intravenously. Receive the blood sample, wipe the surface with a dust-free cloth soaked in 75% alcohol for disinfection, and put it into a biological safety cabinet.

[0156] 2.2 Take 15mL blood sample and transfer it to a clean and sterile 50mL centrifuge tube.

[0157] 2.3 Add normal saline (NS) for injection to a volume of 30 mL in a 50 mL centrifuge tube, and mix the blood cells evenly.

[0158] 2.4 Transfer 15mL GE Ficoll Lymphocyte Separation Solution to a clean and sterile 50mL centrifuge tube.

[0159] 2.5 After the Ficoll is equilibrated at room temperature, ...

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Abstract

The invention relates to the technical field of immune cell therapy, in particular to an initial immune cell activation and amplification system and application thereof. The invention provides an initial immune cell activation and amplification system. A TGF-beta antagonist is added into an activation culture medium, TGF-beta R on the surface of a T cell in a peripheral blood mono-nuclear cell is competitively combined, the combination of TGF-beta and TGF-beta R is blocked, a TGF-beta mediated signal channel is inhibited, the quantity and function of abnormally increased Treg cells are reduced, the functional activity and amplification capacity of T effector cells and antigen presenting cells are up-regulated, the insufficient number and function decline of initial immune cells caused by negative immune regulation mechanisms such as TGF-beta up-regulated Treg cells are avoided, and high-quality and high-quantity excellent seed cells are provided for later successful preparation of targeted immune cells. The invention also provides a method for activating and amplifying initial immune cells and a method for preparing targeted immune cells.

Description

technical field [0001] The invention relates to the technical field of immune cell therapy, in particular to an initial immune cell activation expansion system and its application. Background technique [0002] Adoptive cell therapy (ACT) of cancer relies on the production of anti-tumor immune cells with tumor-recognizing activity. The advantage of this therapy is that it endows immune cells with specific anti-tumor functions through in vitro operations such as antigen loading and gene modification. , and further activate and expand the culture in vitro to obtain a sufficient amount of immune effector cells that kill tumor cells, and the infused immune effector cells can survive as long as possible in vivo and exert a sustained anti-tumor effect. For patients who are willing to accept adoptive cell therapy, after signing the informed consent form, the index screening and immune status evaluation before routine treatment will be performed. According to the T lymphocyte count ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/078A01N1/02
CPCC12N5/0634A01N1/0221C12N2501/515C12N2501/51C12N2501/2302C12N2501/2307C12N2501/2315C12N2501/15
Inventor 翟志敏
Owner THE SECOND AFFILIATED HOSPITAL OF ANHUI MEDICAL UNIV
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