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Cell membrane tumor vaccine and preparation method and application thereof

A technology of tumor vaccines and cell membranes, applied in the field of biological tumor vaccines, can solve the problems of difficult preservation of immune cells, risk of carcinogenesis, and long time consumption

Inactive Publication Date: 2019-08-06
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The main disadvantages of the above methods: (1) The screening and identification of tumor antigens is complicated, costly, and time-consuming, which cannot meet the current urgent need for tumor vaccines; (2) Tumor lysates contain tumor genetic materials, which have potential Carcinogenic risk; (3) It is difficult to preserve the reinfused immune cells

Method used

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  • Cell membrane tumor vaccine and preparation method and application thereof
  • Cell membrane tumor vaccine and preparation method and application thereof
  • Cell membrane tumor vaccine and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] [Example 1] Preparation of Nano Cell Membrane Tumor Vaccine (NP@FMs)

[0054] 1. Inactivation of tumor cells

[0055] The tumor cells were inoculated on a cell culture dish, and 10 ml of 1640 medium containing 10% fetal bovine serum and 1% double antibody (penicillin-streptomycin, 10000 units / ml) was added. The dishes were incubated in a humidified incubator at 37°C containing 5% carbon dioxide. After a period of time, observe under the microscope, and when the cells are full, digest the cells from the culture dish with trypsin, collect them by centrifuging at a speed of 500g for 5 minutes, and wash them three times with PBS. Record the number of cells with a cell counter. The collected cells were treated with 20% ethanol solution at 4° C. for 15 minutes to inactivate tumor cells, and were centrifuged at 500 g for 10 minutes to collect the cells. Redisperse the cells with PBS, centrifuge at 500g for 10 minutes, repeat the above PBS dispersion and centrifugation three...

Embodiment 2

[0064] [Example 2] Preparation of Nano Cell Membrane Tumor Vaccines (NP@FMs)

[0065] The preparation of inactivated tumor cells and BMDCs in this example is derived from Example 1.

[0066] 1. Fusion inactivated tumor cells and dendritic cells to obtain fusion cells;

[0067] The dendritic cells dispersed in PBS and the inactivated tumor cells dispersed in PBS were evenly mixed at a ratio of 10:1, centrifuged at 500 g for 10 minutes, and the supernatant was discarded as much as possible. Within 60 seconds, 2 ml of serum-free 1640 medium containing 50% polyethylene glycol (molecular weight: 2000) and 10% dimethyl sulfoxide that had been warmed to 40° C. was added dropwise. Incubate at 40°C for 1 minute, slowly add serum-free 1640 medium to 50 ml, and terminate the fusion process. Centrifuge at a speed of 500g for 10 minutes to separate the cells, and then disperse the cells in a medium containing 10% fetal bovine serum, 1% double antibody (penicillin-streptomycin, 10000 units / ...

Embodiment 3

[0071] 120 mg of tetrakis (4-carboxyphenyl) porphin, 360 mg of ZrOCl 2 , and 3.36 g of benzoic acid were dissolved in 60 ml of N,N-dimethylformamide. Stirring at 90° C. for 10 hours, the mixed solution was centrifuged at 12000 rpm for 30 minutes, and washed three times with N,N-dimethylformamide to remove unreacted raw materials. The obtained NPs were stored in N,N-dimethylformamide solution. Before coating, replace N,N-dimethylformamide with ultrapure water by centrifugation (12000rpm for 30 minutes). Mix 10 mg / mL of confluent cell membranes and 10 mg / mL of nanoparticles, and sonicate the mixed solution for 1 min in an ice-water bath. The mixed solution is centrifuged at a speed of 10,000 g for 30 minutes to obtain the nano-cell membrane tumor vaccine. [Example 3] Preparation of Nano Cell Membrane Tumor Vaccines (NP@FMs)

[0072] The preparation of inactivated tumor cells and BMDCs in this example is derived from Example 1.

[0073] 1. Fusion inactivated tumor cells and ...

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Abstract

The invention discloses a cell membrane tumor vaccine and a preparation method and an application thereof. For the first time, exogenous tumor antigens and adoptive cells are not used, a cell fusion technology is used to fuse dendritic cells and tumor cells, and after culture, the whole tumor antigen and costimulatory molecules are expressed on the cell membrane of the fused cells. A nano cell membrane vaccine prepared by coating the nanoparticle with the cell membrane subjected to peeling of the fused cell can directly activate the T cell like the antigen presenting cell, can also be recognized by the dendritic cell, processed and presented to the T cell, and indirectly activates the T cell. Two T cells are combined for activation of pathways, which can cause the strong anti-tumor immuneresponse. A cell membrane tumor vaccine strategy that mimics the tumor cells and the antigen presenting cells enables the development of the vaccines against a variety of tumor types, and has good biocompatibility and system safety. Therefore, the cell membrane vaccine is very suitable for the field of tumor vaccines.

Description

technical field [0001] The invention belongs to the field of biological tumor vaccines, and relates to a cell membrane tumor vaccine, a preparation method and application thereof. Background technique [0002] In recent years, many scholars have carried out research on tumor vaccines. By injecting vaccines, training the autoimmune system to monitor and eliminate systemic tumors, and achieving a memory effect, it is possible to monitor and prevent the occurrence of tumors for a long time. However, tumor cells undergo antigenic or other changes that allow them to escape elimination by the immune system, such as tumor cells activating downregulation of certain types of tumor antigens to evade an immune response. Therefore, tumor vaccines containing as many tumor antigens as possible can avoid immune escape caused by tumor cells down-regulating the expression of certain tumor antigens. However, the number of tumor antigens known so far is very limited. In addition, combining ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61P35/00C12N5/0784C12N5/09C12N5/16C12N13/00
CPCA61K39/0011A61K2039/585A61P35/00C12N5/0639C12N5/0693C12N5/16C12N13/00
Inventor 冯俊张先正刘文龙邹梅珍
Owner WUHAN UNIV
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