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Tumour universal type fibroblast vaccine as well as preparation method and applications thereof

A fibroblast and tumor vaccine technology, which is applied in the field of basic fibroblast growth factor-activated fibroblasts and their preparation, can solve the problems of weak immunogenicity and poor specificity of vaccines, and achieves prolonged survival, convenient material collection, Growth and metastatic effects

Active Publication Date: 2017-03-29
WEST VAC BIOPHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the existing anti-tumor vaccines are vaccines targeting tumor cells themselves or tumor-associated antigen-presenting cells (such as DCs (dendritic cells). It is worth noting that the existing vaccines have weak immunogenicity and poor specificity. The disadvantages are still evident

Method used

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  • Tumour universal type fibroblast vaccine as well as preparation method and applications thereof
  • Tumour universal type fibroblast vaccine as well as preparation method and applications thereof
  • Tumour universal type fibroblast vaccine as well as preparation method and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Embodiment 1 Preparation of anti-tumor fibroblast vaccine of the present invention

[0050] All of the following operations were performed under sterile conditions:

[0051] 1) The skin of the back and abdomen of the mouse was surgically excised, placed in PBS containing 100 U / ml ampicillin and 100 mg / ml-streptomycin sulfate, rinsed repeatedly with PBS, and the fat and connective tissue were discarded. Cut the skin to 1mm 3 Add the left and right small pieces into DMEM containing 1 mg / ml type I collagenase, centrifuge at 800g for 3 minutes at 37°C for 2-3 hours, and collect the cells. After 2 washes with DMEM, adjust the cell density to 2-4×10 per ml with DMEM containing 15% fetal bovine serum. 5 Cells were seeded at 75cm 2 Culture flask, 37°C, 5% CO 2 , cultivated under 90% humidity conditions.

[0052] 2) Subculture of skin fibroblasts: After the cultured skin fibroblasts grow into a monolayer, rinse once with PBS without calcium and magnesium ions, add PBS diges...

Embodiment 2

[0055] The characteristics of embodiment 2 fibroblast vaccine of the present invention

[0056] In order to clarify that the present invention has obtained activated fibroblasts, we use the Western Blot method to detect the expression of biological markers of activated fibroblasts. figure 2 It can be seen that the fibroblasts stimulated by human recombinant bFGF highly expressed myofibroblast activation proteins - α-SMA and FAP, while the expression levels of fibroblasts that were not added with human recombinant bFGF during culture were very low. It shows that the fibroblasts obtained through the treatment in Example 1 are activated fibroblasts.

Embodiment 3

[0057] Embodiment 3 Anti-tumor effect of fibroblast vaccine of the present invention

[0058] (1) Experimental grouping

[0059] The present invention uses three kinds of tumor models to verify its anti-tumor effect, including CT26 colon cancer model, LL2Lewis lung cancer model and MethA fibrosarcoma model, each tumor model is divided into two groups: control group (NS group) and fibroblast vaccine therapy group.

[0060] (2) Immunization program

[0061] 6-8 weeks old female Balb / c mice were used for CT26 and MethA tumor models, and 6-8 weeks old female C57BL / 6J mice were used for Lewis lung cancer model, and were randomly divided into groups, 10 mice in each group. The mice in the treatment group were subcutaneously inoculated with fibroblast anti-tumor vaccine 3×10 6 Cells / 200 microliters, the second immunization was carried out 1 to 2 weeks after the first immunization, and the site and dose were the same as the first time. And observe the general state of the mice, wh...

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Abstract

The invention belongs to the field of biological medicine, and relates to basic fibroblast growth factor activated fibroblast as well as a preparation method and an application thereof. The invention aims at solving the technical problem for providing a new selection for tumour treatment. The invention provides a tumour universal type fibroblast vaccine, and a main active component of the vaccine is inactivated basic fibroblast growth factor activated fibroblast. The invention aims at providing the novel cell vaccine for researching and developing tumour fibroblast; and the vaccine can inhibit a plurality of tumours, and is used as a universal antineoplastic vaccine for control of colorectal carcinoma, lung cancer, fibrosarcoma and the like.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals, and relates to a fibroblast activated by basic fibroblast growth factor, a preparation method and application thereof. Background technique [0002] The tumor microenvironment plays a key role in tumor growth, invasion and metastasis, and fibroblasts play a prominent role in tumor formation and development. Existing studies have found that fibroblasts are in an activated state in a variety of malignant tumors, and abnormally express and paracrine a variety of cytokines, which act on tumor cells and tumor stromal cells to promote tumor growth and invasion; activate endothelial cells and Epithelial cells thus promote angiogenesis; promote the malignant transformation of epithelial cells, and promote the occurrence and evolution of tumors. Therefore, fibroblasts can be used as new targets for the treatment of tumors. [0003] Fibroblasts are normally in a quiescent state. During tissue remodelin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61K48/00A61P35/00A61P35/02
CPCY02A50/30
Inventor 石华山王震玲杨莉魏于全
Owner WEST VAC BIOPHARMA CO LTD
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