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Method for manufacturing immunotherapy vaccine

A technology for immunotherapy and vaccines, applied in biochemical equipment and methods, blood/immune system cells, medical preparations containing active ingredients, etc., can solve the problems of low fusion efficiency of dendritic cells and cancer cells

Pending Publication Date: 2021-03-09
CYTIX INC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] Patent Document 1 is proposed as a prior art for solving existing problems such as low fusion efficiency of dendritic cells and cancer cells

Method used

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  • Method for manufacturing immunotherapy vaccine
  • Method for manufacturing immunotherapy vaccine
  • Method for manufacturing immunotherapy vaccine

Examples

Experimental program
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Embodiment Construction

[0041] Embodiments of the present invention are described below. The scope of rights of the present invention is not limited by this example.

[0042] (A) Extraction of exosomes from cell culture supernatant

[0043] Step 1: After culturing for 2 days in FCS-supplemented medium from which exosomes were removed, extraction was performed from the culture supernatant. For extraction, miRCURY Exosome Cell / Urine / CSF Kit (TAKARA BioCat. 300102) was used as an isolation kit.

[0044] Step 2: Measure the number of exosome particles extracted in step 1.

[0045] Step 3: Exosomes are inactivated by heat treatment (56° C., 30 minutes).

[0046] Exosomes are easily disintegrated by heating the cell membrane. Since the cell membrane disintegration cannot fuse with the dendritic cells, the upper limit temperature and the upper limit time of the heat treatment are determined by changing the heating conditions. The method of determination is based on observation of the membrane structure...

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Abstract

The invention provides a method for manufacturing vaccine used for immunotherapy of cancer. If exosomes are left at room temperature for a certain period of time or more, its membrane structure collapses and cannot fuse with collapsed dendritic cells. Therefore, it is necessary to perform heat treatment under conditions that maintain the exosome membrane structure while promoting peptide formationof proteins, and this condition is 56 DEG C or less and 30 minutes or less. At a temperature condition higher than this, protein is denatured and cannot retain its antigenicity. Therefore, exosomes separated from body fluids such as cancer cells or blood of a cancer patient are subjected to heat treatment under conditions of 56 DEG C or lower and 30 minutes or lower, thereby resulting in promoting inactivation of protein enzymes in exosomes and inactivating protein degrading enzymes and lipolytic enzymes. Then, the exosomes are introduced into or co-cultured with dendritic cells derived fromblood of a cancer patient or a healthy person.

Description

technical field [0001] The present invention relates to a method for preparing a dendritic cell vaccine used in immunotherapy using exosomes of cancer cells and dendritic cells. Background technique [0002] For immunotherapy, there is a therapy (acquired immunity) that uses specific proteins produced by cancer cells as markers to present specific antibodies to generate immune cells (CTL) that attack cancer cells, and recognizes foreign proteins to eliminate foreign substances ( natural immunity) two therapies. [0003] In conventional immunotherapy, dendritic cells (DC: dendritic cells) isolated from blood mononuclear cells of a patient are fused with cancer cells (including precancerous cells) obtained from the patient, and the fused cells are isolated or Administration is by subcutaneous injection or the like together with cytokines that stimulate cytotoxic T cell responses, humoral immune responses. [0004] In the case of this conventional immunotherapy, the fusion ef...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61P35/00
CPCA61K39/4622A61K39/4615A61K39/4644A61P35/00A61K2239/31C12N5/0639C12N5/0693C12N2502/1121C12N2523/00C12N2502/1114C12N13/00
Inventor 佐久间真俊大园三千代冈惠理子
Owner CYTIX INC
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