A kind of radioresistance lung cancer whole cell vaccine and its preparation method and application

A whole cell vaccine and lung cancer cell technology, applied in the field of radioresistant lung cancer whole cell vaccine and its preparation, can solve the problem of inability to improve the immune function of T lymphocytes in C57BL/6 mice, and achieve long-lasting specific anti-tumor effect, inhibiting Significant effect on tumor and inhibiting the growth of lung cancer tumor in mice

Active Publication Date: 2018-05-25
JINSHAN HOSPITAL FUDAN UNIV
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  • Summary
  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Conclusion: The use of Lewis tumor whole cell vaccine can not improve the immune function of T lymphocytes in C57BL / 6 mice, but can improve the immune function of B lymphocytes, but can not make mice obtain effective immune protection
[0004] So far, there is no whole-cell vaccine against lung cancer with strong immunogenicity, significant anti-tumor effect and long-lasting effect

Method used

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  • A kind of radioresistance lung cancer whole cell vaccine and its preparation method and application
  • A kind of radioresistance lung cancer whole cell vaccine and its preparation method and application
  • A kind of radioresistance lung cancer whole cell vaccine and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Establishment of the radioresistant lung cancer cell line of the present invention

[0044] 1 Cell culture and irradiation method

[0045] LLC Lewis lung cancer cell line (abbreviated as LLC cells, purchased from ATCC) was placed in DMEM medium containing 10% fetal bovine serum at 37°C and 5% CO 2 The cells were cultured in an incubator with saturated humidity, and the cells were subcultured to the exponential growth phase. 6MV X-rays were irradiated with a linear accelerator, the source-to-skin distance was 100cm, and the surface was covered with 1cm-thick tissue compensation glue.

[0046] 2 Establishment of radioresistant cell line R-LLC

[0047] Take LLC cells in the exponential growth phase, digest them into a single cell suspension and inoculate them on 25cm 2 In the culture flask, after the cells grow to the exponential growth phase, the pre-irradiation field is 7cm×7cm, 6MV X-rays, the absorbed dose rate is 200cGy / min, 5Gy is irradiated, and then th...

Embodiment 2

[0068] Example 2 Preparation of the radioresistant lung cancer whole cell vaccine of the present invention

[0069] For the R-LLC cells obtained by repeated radiation irradiation for routine passage>10 passages, the R-LLC cells in the exponential growth phase were collected, the culture supernatant was collected, counted on the counting plate, and the cell concentration was adjusted to 5.0×10 with the culture supernatant. 6 cells / ml, first inactivate the cells by repeated freezing and thawing (freeze in liquid nitrogen for 10 min, then quickly transfer to a 37°C water bath for 10 min, repeat four times), and crush them with an ultrasonic cell pulverizer on ice (power 130W / 20KHz, time 3s , 10-15 times) cells, the vortex shaker fully oscillated to prepare tumor seedlings, and the final concentration of tumor seedlings was 1.0×10 6 Individual tumor cells / 0.2ml.

Embodiment 3

[0070] Example 3 Anti-tumor effect of CpG ODN1826 combined with radioresistant lung cancer whole cell vaccine of the present invention

[0071] 1 method

[0072] 1.1 Immunization method and experimental grouping

[0073] There were 20 mice in each group. The mice in each group were first immunized with CpG ODN1826 (purchased from Shanghai Sangon Bioengineering Co., Ltd., product number: 9103288756) three times, and then immunized with tumor vaccine three times. Repeated immunization every day, divided into the following six groups according to different immunization methods:

[0074] (a) Negative control group (Control): Each mouse was subcutaneously injected with 0.15ml of PBS as the CpGODN1826 drug control, and then subcutaneously injected with 0.2ml of serum-free culture medium at multiple points as the tumor vaccine control;

[0075] (b) CpG ODN group (CpG): Each mouse was subcutaneously injected with CpG ODN1826 0.15 mg each time, and then subcutaneously injected with s...

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Abstract

The present invention relates to a radiation-resistant lung cancer whole-cell vaccine and its preparation method and application. Specifically, the present invention establishes a radiation-resistant lung cancer cell line with significantly improved radiation resistance. The preservation number is CCTCC C201537. The cell line is used to A lung cancer whole-cell vaccine was prepared, and animal experiments confirmed that the lung cancer whole-cell vaccine had the effects of significantly inhibiting the growth of mouse lung cancer tumors, promoting apoptosis of transplanted tumor cells, and reducing the expression of PDL-1 protein in immune escape of transplanted tumors, and combined with CpG ODN The anti-tumor effect is more significant, and the tumors of some mice completely disappear, and a durable and specific anti-tumor effect is obtained. The invention provides new ideas and methods for the treatment of lung cancer and the prevention of postoperative metastasis and recurrence.

Description

technical field [0001] The invention relates to a tumor cell vaccine, in particular to a radiation-resistant lung cancer whole cell vaccine and its preparation method and application. Background technique [0002] Lung cancer is one of the common malignant tumors in many countries, and has become the first cause of human death due to tumors. The latest survey shows that it has occupied the first place in the incidence of malignant tumors in men and the mortality rate of malignant tumors in men and women in China. Conventional treatment methods include surgery, radiotherapy and chemotherapy, but the curative effect is not satisfactory at present. These treatments can easily damage normal tissues while killing or eliminating tumor cells, especially the immune system, which plays an important role in the body's anti-tumor defense. The immune escape of tumor cells and the dysfunction of the immune system in patients promote the occurrence and development of tumors. Therefore, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09A61K39/00A61P35/00A61P37/04A01K67/027C12Q1/02
Inventor 乔田奎庄喜兵燕丽邢娜张琪
Owner JINSHAN HOSPITAL FUDAN UNIV
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