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Method for enhancing anti-tumor effect of immune cells

A technology of immune cells and cells, which is applied in the field of cell culture technology and immunotherapy, and can solve the problems that the therapeutic effect needs to be further improved

Pending Publication Date: 2022-03-29
BEIJING CANCER HOSPITAL PEKING UNIV CANCER HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] T cell immunotherapy mainly includes chimeric antigen receptor gene-modified T cells (CAR-T), T cell receptor gene-modified T cells (TCR-T) and tumor infiltrating T cells (TILs). The therapeutic area has shown certain clinical therapeutic effect, but in most solid tumors, its therapeutic effect needs to be further improved

Method used

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  • Method for enhancing anti-tumor effect of immune cells
  • Method for enhancing anti-tumor effect of immune cells
  • Method for enhancing anti-tumor effect of immune cells

Examples

Experimental program
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Effect test

Embodiment 1

[0054] The acquisition of embodiment 1TILs

[0055] After the tumor tissue of the tumor patient is surgically removed, the tumor tissue is chopped into small pieces of 1-2 mm, and each small piece of tumor tissue is placed in a well of a 24-well cell culture plate, and then T cell culture medium is added.

[0056] The T cell culture medium was X-VIVO serum-free medium (Lonza, USA) supplemented with IL2 (final concentration 50U / ml; Peprotech, USA), OKT3 antibody (final concentration 50ng / ml; ACRO, USA) and anti- CD28 antibody (final concentration 1ug / ml; T&L Biotechnology, China). The small pieces of tumor tissue were cultured in a cell culture incubator until final TILs were obtained.

Embodiment 2

[0057] Example 2 The expression of TILs depletion indicators treated with spermidine decreased significantly, and the proliferation ability could be enhanced

[0058] The TILs obtained in Example 1 were divided into two groups, the control group (Control) was cultured for 7 days through the T cell culture medium mentioned in Example 1, and the spermidine group (spermidine) was treated with spermidine (10uM) (Cayman Chemical, The T cell culture medium mentioned in Example 1 of USA) was cultivated for 7 days.

[0059] The TILs of the two groups were stained by flow cytometry antibody and the data were analyzed. The results of the study found that the ratios of PD1, TIM3 and LAG3 expressed by the TILs of the spermidine group alone were about 20%, 60%, and 20%, respectively, while the TILs of the control group alone expressed The ratios of expressing PD1, TIM3 and LAG3 were about 30%, 70%, and 30%, respectively. It can be seen that the ratio of TILs expressing immunosuppressive re...

Embodiment 3

[0060] Example 3 The anti-tumor ability of TILs treated with spermidine in vivo and in vitro is significantly enhanced

[0061] The TILs obtained in Example 1 were divided into two groups, the control group (Control) was cultured for 7 days through the T cell culture medium mentioned in Example 1, and the spermidine group (spermidine) was added with spermidine (10uM) (Cayman Chemical , USA) T cell culture medium described in Example 1 for 7 days.

[0062] 1. TILs treated with or without spermidine were compared with 10 5Autologous lung cancer tumor cells were co-incubated with an effector-target ratio of 1:1 for 24 hours and then the supernatant IFN-γ concentration was detected (for the detection method, please refer to the literature: J Immunother Cancer. 2019 Aug 28; 7(1):232.). The results showed that the IFN-γ secreted by spermidine-treated TILs was significantly higher than that of untreated TILs ( figure 2 A and B).

[0063] 2. TILs treated with spermidine or not wer...

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Abstract

The invention relates to a method for enhancing the anti-tumor effect of immune cells, and belongs to the technical field of cell culture. In vitro culture amplification of 7-10 days is generally needed before CAR-T, TCR-T and TILs of tumor patients are used for treatment, however, it is found that by adding spermidine into a culture medium for in vitro culture, the functional effects of the CAR-T, the TCR-T and the TILs can be recovered, the multiplication capacity is improved, the IFN-gamma secretion level is improved, and the in-vivo and in-vitro autologous tumor cell killing capacity is remarkably enhanced; therefore, the CAR-T, TCR-T and TILs treated in vitro by adopting spermidine have a better cancer treatment effect. A new method is provided for tumor treatment, and the clinical application prospect is good.

Description

[0001] This application is a divisional application of a patent application with an application date of August 13, 2021, an application number of 2021109274647, and an invention titled "A Method for Enhancing the Anti-tumor Effect of Immune Cells". technical field [0002] The invention relates to the field of cell culture technology and immunotherapy technology, in particular to a method for enhancing the anti-tumor effect of immune cells. Background technique [0003] Spermidine, also known as spermidine trihydrochloride, is a kind of polyamines. Polyamines are small, fatty, polycationic (-NH 3+ ) of biomolecules. There are mainly four polyamines in mammals: spermine, spermidine, putrescine and cadaverine. Spermine belongs to tetramines, spermidine belongs to triamines, putrescine and cadaverine are diamines, and the different numbers of amino groups make them have different physiological characteristics. Spermidine is a low molecular weight aliphatic carbide containing...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783
CPCC12N5/0636C12N5/0638C12N2501/999C12N2501/2302C12N2501/515C12N2501/51C12N2500/90
Inventor 陆哲明张超亭
Owner BEIJING CANCER HOSPITAL PEKING UNIV CANCER HOSPITAL
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