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Application method of tumor cell-derived exosome antigen in DC vaccine

A tumor cell and tumor vaccine technology, which is applied in the application field of tumor cell-derived exosome antigens in DC vaccines, can solve the problems of low direct uptake efficiency of antigens, etc., to enhance the body's anti-tumor immunity, enhance cytotoxic effect, good stability

Pending Publication Date: 2019-12-24
SHENZHEN PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the low efficiency of direct uptake of free antigen by DC, the construction of an efficient and safe DC vaccine antigen presentation carrier is a key problem to be solved urgently, and it is also an important task facing the current DC vaccine industry

Method used

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  • Application method of tumor cell-derived exosome antigen in DC vaccine
  • Application method of tumor cell-derived exosome antigen in DC vaccine
  • Application method of tumor cell-derived exosome antigen in DC vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1, Preparation of Exosomes in the Culture Supernatant of Lung Tumor A549

[0058] 1. Preparation of exosome-free FBS

[0059] Dissolved and inactivated FBS (Gibco, USA) was centrifuged at 12 000 g at 4°C for 30 min to remove impurities with larger molecular weights. Use a syringe to slowly inject the serum supernatant into the ultracentrifuge tube, and remember not to generate air bubbles. After the supernatant is added to the bottleneck of the centrifuge tube, use an electric fuser to melt the centrifuge tube mouth. The centrifuge tubes that have been melted are placed diagonally and symmetrically on an ultracentrifuge, and centrifuged at 150,000 g for 12 hours at 4°C. After centrifugation, cut off the seal, aspirate the supernatant, and filter the supernatant with a 0.22 μm syringe filter.

[0060] The filtered supernatant (FBS with exosomes removed) was subpackaged and stored at -20°C for future use.

[0061] 2. Obtain tumor cell culture supernatant in ser...

Embodiment 2

[0080] Example 2. Application of exosomes in tumor cell culture supernatant in DC vaccine

[0081] 1. A549 cell culture supernatant-derived exosomes activate DC

[0082] 1. Induction and culture of human peripheral blood-derived dendritic cells (DC)

[0083] (1) Collect 25ml of peripheral blood from healthy volunteers in vitro, pipette the peripheral blood up and down to mix, 800g, centrifuge for 5min, the upper layer is light yellow plasma layer, and the remaining liquid is concentrated blood cells.

[0084] (2) Put the upper layer of plasma obtained in step (1) into a centrifuge tube and seal it with a parafilm, inactivate it in a water bath at 56°C for 30 minutes, and take the supernatant after centrifugation for later use.

[0085] (3) Aspirate the blood cells obtained in step (1) and slowly transfer the diluted blood cells into a centrifuge tube filled with lymphocyte separation medium (GE, USA) at 1 cm from the slope of the separation medium. The transferred blood cell...

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Abstract

The invention discloses an application method of a tumor cell-derived exosome antigen in a DC vaccine. The invention discloses a preparation method of a tumor vaccine, which comprises the following step: sensitizing DC cells by using a tumor cell exosome to obtain the tumor vaccine. The tumor cell supernatant exosome provided by the invention has good stability, low toxicity and good cell compatibility, and is convenient to operate and apply and popularize in clinical tumor immunotherapy, and the method is a simple, feasible and practical application technology and method. The problems that inexisting DC vaccine construction, antigen sources are lacked, compatibility is poor, instability is caused, and an external DC antigen carrier has biotoxicity and is difficult to operate are solved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for applying tumor cell-derived exosome antigens in DC vaccines. Background technique [0002] As we all know, malignant tumors are the primary cause of serious threats to human health, and the development of high-efficiency and low-side-effect treatment models is crucial to improving the survival rate and quality of life of patients. In the past ten years, although the specificity and safety of anti-tumor drug therapy have been greatly improved, tumor invasion, metastasis, recurrence, and patient resistance to conventional drug therapy are still the root causes of tumors that cannot be effectively controlled. Therefore, how to treat cancer in the body Inducing a durable anti-tumor effect and avoiding recurrence after treatment are still bottlenecks that need to be solved urgently. [0003] As a new tumor treatment mode, tumor immunotherapy mainly generates anti-tumor immun...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61P35/00
CPCA61K39/0011A61P35/00
Inventor 王策李富荣黄雪陈尚
Owner SHENZHEN PEOPLES HOSPITAL
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