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Tumour vaccine and preparation method thereof

A technology for tumor vaccines and tumor cells, applied in the fields of tumor vaccines and their preparation, exosomes tumor vaccines and their preparations, capable of solving rare, destructive, and negative problems, achieving effective killing, less toxic and side effects, and safe use

Inactive Publication Date: 2014-05-28
FIRST HOSPITAL AFFILIATED TO GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although there are many related studies on tumor vaccines at home and abroad, so far few breakthroughs have been made. The main problems are: the technology of introducing DNA into specific cells for expression is not yet mature, and the use of exogenous The safety of DNA has not yet been resolved; due to the high heterogeneity of tumor cells, multiple tumor cells belonging to the same type of tumor can express different antigens, and T cells activated by a tumor antigen can only kill a part of the tumor cells , tumor cells that do not express this antigen cannot be killed; although tumor cell vaccines can contain almost all tumor antigens, current studies have shown that their role in activating T cells is limited, and the effect of using them as a vaccine is not ideal
[0008] However, in recent years, some related experiments have shown that some tumor-derived exosomes can inhibit or even destroy immune cells that play a role in tumors, such as down-regulating the expression of some NK receptors, affecting the activation of some innate immune cells in tumor immunity , and some can significantly inhibit IL-2 to inhibit the proliferation of human lymphocytes, thus playing some negative roles in tumor immunotherapy
These tumor-derived exosomes may be the key factor for tumor tissue to escape the body's immune system clearance, which brings many difficulties and challenges to tumor immunotherapy

Method used

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  • Tumour vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Cell Culture

[0045] Experimental materials: H22 cell line, 10% fetal bovine serum, penicillin and streptomycin mixture (Beijing Tigemei Company).

[0046] Experimental method: the H22 cell line was cultured in a culture medium containing 10% fetal bovine serum, penicillin 100IU / mL, and streptomycin 100×μg / mL, at 37°C and 5% CO 2 Culture in the incubator, when the cells grow to the logarithmic phase, press 3×10 6 / 100ml inoculation.

Embodiment 2

[0047] Embodiment 2 drug treatment

[0048] Experimental materials: hydralazine, SAHA (both purchased from sigma company).

[0049] Experimental grouping: the volume of cell culture medium is strictly consistent in each group, blank control group, exosomes control group (no drug addition group), exosomes experimental group 1 (adding hydralazine), exosomes experimental group 2 (adding drug SAHA), exosomes Experimental group 3 (combined administration of hydralazine and SAHA).

[0050] Experimental method: in the control group, no drug was added after inoculation, and 150ml of the supernatant was collected as a control after 24 hours; -6 mol of hydralazine treatment, respectively collected 150ml of culture supernatant 24h after dosing; dosing group 2, 24h after inoculation with a concentration of 1 × 10 -6 mol / L SAHA treatment, respectively collected 150ml of culture supernatant 24h after dosing; dosing group 3, 24h after inoculation with 1 × 10 -6 mol / L of hydralazine and 1×...

Embodiment 3

[0051] The separation and purification of embodiment 3 exosomes

[0052] Experimental equipment: HMAC-CP7OG low-temperature ultra-high-speed centrifuge, 100KU MW C0Millipore Amicon high recovery rate high flow rate tangential flow ultrafiltration centrifuge tube (Millipore Company).

[0053] Experimental method: Centrifuge the collected cell culture supernatants of the experimental group and the control group at 300g for 10min to remove the cells, and take the supernatant; centrifuge at 1500g for 30min to remove the cell debris, collect the supernatant, and concentrate it through a 100kUMWCO Centriplus centrifugal ultrafiltration tube Ultrafiltration, centrifugation at 1500g for 30min to obtain 6ml of concentrated solution, transfer the separated and purified concentrated solution to a 1.5ml centrifuge tube, and ultracentrifuge at 100kg for 60min at 4°C with a horizontal rotation angle to obtain exosomes.

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Abstract

The invention discloses a tumour vaccine and a preparation method thereof. According to the invention, hydralazine and SAHA are adopted to jointly treat tumor cells so as to obtain an exosmoes tumour vaccine. The invention further discloses a preparation method of the tumour vaccine, which comprises the steps that the hydralazine cooperates with the SAHA to treat the tomour cells, and separate and purify exosomes excreted by the tomour cells. The tumour vaccine disclosed by the invention improves the curative effect of the exosomes tumour vaccine, and has important clinic application value.

Description

technical field [0001] The invention relates to a vaccine, in particular to a tumor vaccine and a preparation method thereof, more specifically to an exosomes tumor vaccine and a preparation method thereof. Background technique [0002] In recent years, as a disease that seriously threatens people's lives, tumor treatment has become the subject of research by many researchers. With the rapid development of immunology, cell biology and molecular biology, tumor surgery, radiotherapy After the three major therapies of chemotherapy and chemotherapy, biological therapy of tumor has become the fourth major therapy. Tumor biotherapy includes immunotherapy, gene therapy, stem cell therapy, induction of tumor cell differentiation and apoptosis, and inhibition of tumor angiogenesis. Immunization methods to treat tumors have attracted much attention because of their low toxicity, among which the research and development of tumor vaccines is the focus of attention. [0003] Tumor vacc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61P35/00C12N5/09
Inventor 肖文华董伟伟
Owner FIRST HOSPITAL AFFILIATED TO GENERAL HOSPITAL OF PLA
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