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Method for preparing antigen composition capable of targeting glioma cells and glioma stem cells, and vaccine containing the antigen composition

A technology for glioma stem cells and glioma cells, which is applied in the application field of medicines, can solve the problems that antigen-presenting cells cannot quickly identify and activate effector cells, cannot effectively simulate the antigen-presenting process, and cannot obtain antigens, etc. To achieve the effect of improving the utilization rate in vivo, inhibiting the growth of glioma, and enhancing the strength of immune response

Inactive Publication Date: 2017-09-19
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The prior art only discloses how to obtain brain glioma stem cells by separation and sorting, but since all glioma stem cells cannot be obtained by sorting glioma stem cell marker proteins (such as CD133, etc.), glioma stem cells can be obtained from postoperative tissues. After the cells are sorted, the number of cells obtained is very small, and a large number of known and unknown glioma stem cell-derived antigens cannot be obtained; the prior art only discloses antigen presentation by contacting glioma stem cell lysates with dendritic cells , this method cannot effectively simulate the antigen presentation process in vivo, and there are still problems that other antigen-presenting cells (such as B cells) do not participate in antigen presentation, and antigen-presenting cells cannot quickly recognize and activate effector cells.

Method used

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  • Method for preparing antigen composition capable of targeting glioma cells and glioma stem cells, and vaccine containing the antigen composition
  • Method for preparing antigen composition capable of targeting glioma cells and glioma stem cells, and vaccine containing the antigen composition
  • Method for preparing antigen composition capable of targeting glioma cells and glioma stem cells, and vaccine containing the antigen composition

Examples

Experimental program
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Effect test

Embodiment l

[0021] Example 1: Preparation of antigen composition containing glioma stem cell lysate and glioma cell lysate

[0022] (1) Obtaining a cell population containing glioma stem cells and glioma cells

[0023]Glioma tissues were obtained from patients with grade IV gliomas. After surgical resection of the tumor tissue, the glioma tissue was quickly placed in Neurobasal (NB) medium containing 5 μg / ml gentamicin, 0.9 μg / ml levomycin, 50 ng / ml EGF and 50 ng / ml FGF, It was then placed in a cryopreservation box and quickly transferred to a GMP laboratory for subsequent processing. In a biosafety cabinet, transfer the tumor tissue to new Neurobasal (NB) medium containing 5 μg / ml gentamicin, 0.9 μg / ml levomycin, 50 ng / ml EGF, and 50 ng / ml FGF, and the tumor Cut the tissue into small pieces, remove the hoof tissue and necrotic tissue, and then further mince the tumor tissue into about 1mm 3 Add sterile saline to the obtained fragmented tissue to form a tissue cell suspension, remove l...

Embodiment 2

[0030] Example 2: Using mouse glioma cell line GL261 to prepare antigen composition containing mouse glioma stem cell lysate and mouse glioma cell lysate

[0031] Resuscitate the mouse glioma cancer cell line GL261 frozen in our laboratory (gifted by Professor Chen Wei, University of Minnesota, USA), culture to the logarithmic growth phase, obtain cell suspension, and wash thoroughly with sterile phosphate buffer saline 2 times, count the cells, resuspend the cells with the complete medium of mouse glioma stem cells, and adjust the cell density to 0.5-1×10 6 / ml, transfer every 10-15ml to a 10cm sterile suspension cell culture dish at 37°C, 5% CO 2 and 5%O 2 Culture in a three-gas incubator and change the medium every 2 days. After 10 days of culture, the medium was changed every 3 days. According to the above culture method, tumor spheres were obtained. When changing the medium, when the diameter of the tumor spheres is greater than 100 μm, the tumor spheres can be blown ...

Embodiment 3

[0039] Embodiment 3: In vivo efficacy detection of the vaccine of the present invention

[0040] In this example, the antigen composition containing glioma stem cell lysates and glioma cell lysates prepared in Example 2, dendritic cells and C-type CpG oligonucleotides (CpG2395, purchased from Invivogen) vaccine was tested for in vivo efficacy; wherein, dendritic cells were isolated from C57BL / 6 mouse bone marrow cells, induced by interleukin 4 (IL-4) and granulocyte-macrophage colony-stimulating factor (GM-CSF), lipopolysaccharide Dendritic cells obtained after (LPS) activation. The vaccine used in this example contains 1000 μg of the antigen composition prepared in Example 2 per milliliter, 4×10 6 dendritic cells and 200 μg C-type CpG oligonucleotides. Each tumor-bearing mouse was injected with 250 μl of vaccine, and 125 μl of vaccine was subcutaneously injected into the shoulders of both sides.

[0041] 50 healthy 6-week-old female SPF-grade C57BL / 6 mice (purchased from B...

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Abstract

The invention provides a method for preparing an antigen composition capable of targeting glioma cells and glioma stem cells. The method comprises the steps of (1) providing isolated cell populations containing the glioma stem cells and the glioma cells; (2) amplifying the glioma stem cells in the cell populations; (3) carrying out freeze-thaw cracking treatment on the amplified cell populations so as to obtain the antigen composition containing glioma stem cell lysates and glioma cell lysates. The invention also provides a vaccine containing the antigen composition, dendritic cells and CpG oligodeoxynucleotide, and application of the vaccine in preparation of medicines for treating glioma. The antigen composition and the vaccine which are provided by the invention can target the glioma cells and the glioma stem cells at the same time, thus being capable of effectively killing the residue glioma stem cells and the glioma cells and further inhibiting the recurrence of the glioma.

Description

technical field [0001] The present invention relates to an antigenic composition containing glioma stem cell lysate and glioma cell lysate, a method for preparing the antigenic composition, a vaccine containing the antigenic composition, and the antigenic composition used for treating glioma Application in tumor medicine. Background technique [0002] At present, the main treatment for glioma is surgical resection, supplemented by radiotherapy and chemotherapy. However, due to the infiltrative growth characteristics of glioma, it is difficult to completely remove the tumor tissue. Due to the special location of the lesion, radiotherapy and chemotherapy are also ineffective. The theory of cancer stem cells and the isolation and identification of glioma stem cells pointed out that the main culprit of glioma recurrence and low survival rate after treatment is that glioma stem cells can self-renew, have high differentiation potential and are insensitive to radiotherapy and chemo...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61K39/395A61P35/00
CPCA61K39/0011A61K39/395A61K2039/5154A61K2039/55561
Inventor 陈京涛朱珊关毅陈卫
Owner JILIN UNIV
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