Radioresistant lung cancer whole-cell vaccine, and preparation method and application thereof

A whole-cell vaccine and lung cancer cell technology, which is applied in the field of radiation-resistant lung cancer whole-cell vaccine and its preparation, can solve the problem of not being able to improve the immune function of T lymphocytes in C57BL/6 mice, achieve a durable and specific anti-tumor effect, reduce Protein expression, the effect of promoting cell apoptosis

Active Publication Date: 2015-09-30
JINSHAN HOSPITAL FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • 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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  • Radioresistant lung cancer whole-cell vaccine, and preparation method and application thereof
  • Radioresistant lung cancer whole-cell vaccine, and preparation method and application thereof
  • Radioresistant lung cancer whole-cell vaccine, and preparation method and application thereof

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 invention relates to a radioresistant lung cancer whole-cell vaccine, and a preparation method and application thereof, and particularly establishes a radioresistant lung cancer cell strain with obviously higher radioresistance, of which the collection number is CCTCC No.C201537. The animal experiment proves that the lung cancer whole-cell vaccine prepared by using the cell strain has the obvious actions on inhibiting mouse lung cancer tumors from growth, promoting transplantation tumor apoptosis and lowering the expression of the transplantation tumor immunologic escape PDL-1 protein; and when being combined with CpG ODN, the lung cancer whole-cell vaccine has more obvious actions on tumor inhibition, and part of mouse tumors completely disappear, thereby obtaining permanent specific antitumor effects. The invention provides a new idea and method for treating lung cancers and preventing postoperative metastasis and relapse.

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 Applications(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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