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Immunocyte medicine containing B cell vaccine loaded with new antigen

A technology of immune cells and immune cell therapy, applied in the field of immune cell therapy drugs, can solve the problems that it is difficult to become an effective means to amplify neoT or ENT, DC vaccine mechanism defects, etc., and achieve good cancer prevention and treatment effects

Pending Publication Date: 2019-09-24
CHINEO MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Question 1: Is it to construct a fixed-target vaccine, or an individualized multi-target neoantigen cancer vaccine?
[0005] Question 2: To construct a polypeptide or RNA vaccine, or to use antigen-presenting cells as the carrier of the vaccine to construct a cellular vaccine?
[0007] Question 3: If it is possible to break through the limitation of the number of antigen-presenting cells and continuously and effectively present new antigens, thereby continuously and effectively activating and expanding new antigen-reactive T cells?
However, DC vaccines have quantitative bottlenecks and mechanism defects, and it is difficult to become an effective means to amplify neoT or ENT

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1: Preparation of B cell vaccine (neoB) and DC cell vaccine (neoDC) loaded with neoantigen

[0019] Isolation of B cells and DC cells from peripheral blood mononuclear cells of tumor patients.

[0020] Isolation and expansion of B cells: Peripheral blood was collected intravenously from patients, PBMCs were separated using Ficoll lymphocyte separation medium, and CD19-positive B cells were sorted and counted using CD19 antibody magnetic beads. Add relevant medium and cytokines to culture and expand B cells, and collect B cells on the 12th day to achieve 10-30 times expansion.

[0021] Isolation, differentiation and maturation of DC cells: Collect peripheral blood from patients intravenously or apheresis, separate PBMCs with Ficoll lymphocyte separation medium and adhere to the wall for 2 hours, take adherent monocytes, add relevant cytokines to promote DC cell differentiation and maturation, After 7-8 days of culture, mature DC cells were collected.

[0022] ...

Embodiment 2

[0023] Example 2: In vitro experiments, the comparison of neoB vaccine and neoDC vaccine activation and enrichment neoT effect

[0024] Isolate tumor infiltrating T lymphocytes (TIL) from autologous tissues of cancer patients, or isolate T cells from peripheral blood mononuclear cells of cancer patients. The mixture of neoB and neoDC prepared from the patient's own cells was co-cultured with T cells for 1 day, and the proportion of CD137+ T cells in all T cells was detected by flow cytometry, that is, neoantigen-reactive T cells (neoT). The results Find:

[0025] The proportion of CD137+ T cells in patient 1: 21.3% for neoB stimulation and 22.1% for neoDC stimulation;

[0026] The proportion of CD137+ T cells in patient 2: neoB-stimulated 6.3%, neoDC-stimulated 6.5%,

[0027] The proportion of CD137+ T cells in patient 3: 3.6% for neoB stimulation, 3.8% for neoDC stimulation,

[0028] The proportion of CD137+ T cells in patient 4: neoB-stimulated 33.7%, neoDC-stimulated 3...

Embodiment 3

[0031] Example 3: Preparation of normal and enhanced neoantigen-reactive T cells

[0032] Isolate tumor infiltrating T lymphocytes (TIL) from autologous tissues of cancer patients, or isolate T cells from peripheral blood mononuclear cells of cancer patients. The mixture of neoB and neoDC prepared from the patient's own cells was used to co-culture with T cells for 1 day, and the activated T cells were screened with CD137 antibody magnetic beads to obtain neoantigen-reactive T cells (neoT).

[0033] The lentiviral vector of the reverse conversion molecule of PD1 / 4-1BB was transferred into neoT, the transfection efficiency was about 60%, and PD1 was sorted out by flow cytometry or magnetic beads + T cells, construct enhanced neoT (ENT), remaining PD1 - T cells are common neoT.

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PUM

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Abstract

The invention provides an immunocyte medicine containing a B cell vaccine (neoB) loaded with a tumor new antigen, and a medicine combination for united use of neoB and tumor specificity T cells. The neoB vaccine can have higher ex vivo expansion properties than a DC vaccine, has higher effect of continuous activating of in vivo T cells under the condition of repeated infusion, and can become a preventive or therapeutic vaccine or an immunocyte medicine to be applied to tumor resisting treatment suitable for any cancer kind. According to the medicine combination for united use of neoB and tumor specificity T cells, disclosed by the invention, the neoB can be used for further stimulating the tumor specificity T cells in bodies, so that the number of the tumor specificity T cells in bodies can be amplified, and the treatment effect is increased.

Description

technical field [0001] The invention belongs to the technical field of biomedicine. Specifically, the invention provides a B cell vaccine loaded with neoantigen and an immune cell therapy drug containing the vaccine. The vaccine or drug can be used for the prevention and treatment of tumors. Background technique [0002] The use of cancer vaccines as an auxiliary or main means of cancer prevention and treatment is a promising direction for cancer prevention and treatment, but there are three problems to be solved for cancer vaccines: [0003] Question 1: Is it to construct a fixed-target vaccine, or an individualized multi-target neoantigen cancer vaccine? [0004] Compared with fixed-target cancer vaccines, individualized neoantigen cancer vaccines are the general trend. [0005] Question 2: To construct a polypeptide or RNA vaccine, or to use antigen-presenting cells as the carrier of the vaccine to construct a cellular vaccine? [0006] Most neoantigen vaccines use pept...

Claims

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Application Information

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IPC IPC(8): C12N5/10A61K39/00A61P35/00
CPCC07K14/4748A61K39/0011A61K35/17A61P35/00
Inventor 谷为岳
Owner CHINEO MEDICAL TECH CO LTD
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