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A kind of initial immune cell activation expansion system and its application

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 obtain CAR-T cells in vitro, so as to ensure the quantity and quality of preparation, avoid insufficient quantity, and avoid The effect of functional decline

Active Publication Date: 2022-01-07
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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  • A kind of initial immune cell activation expansion system and its application
  • A kind of initial immune cell activation expansion system and its application
  • A kind of initial immune cell activation expansion system and its application

Examples

Experimental program
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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. Separation 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 expansion system and its application. The invention provides an initial immune cell activation and expansion system. TGF-β antagonists are added to the activation medium to competitively bind to TGF-βR on the surface of T cells in peripheral blood mononuclear cells to block TGF-β and TGF The combination of ‑βR inhibits the signaling pathway mediated by TGF‑β, down-regulates the number and function of abnormally increased Treg cells, up-regulates the functional activity and expansion ability of T effector cells and antigen-presenting cells, and avoids the up-regulation of Treg cells due to TGF-β Insufficient number and decreased function of "initial immune cells" caused by such negative immune regulation mechanism provide high-quality and high-quantity excellent seed cells for the successful preparation of targeted immune cells in the later stage. The invention also provides a method for activating and expanding 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 Patents(China)
IPC IPC(8): C12N5/078A01N1/02
CPCC12N5/0634A01N1/0221C12N2501/515C12N2501/51C12N2501/2302C12N2501/2307C12N2501/2315C12N2501/15
Inventor 翟志敏高志买李丛王会平
Owner THE SECOND AFFILIATED HOSPITAL OF ANHUI MEDICAL UNIV
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