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Application of histone deacetylase and inhibitor thereof to preparation of tumor whole cell antigen loaded DC (dendritic cell) tumor vaccine

A deacetylase, whole cell antigen technology, applied in the field of tumor vaccines, can solve the problems of reducing the ability of migration, interfering with the antigen presentation process, affecting the differentiation and maturation and migration of DCs, and achieving improved prevention and treatment effects, high prevention and Therapeutic value, important research significance and effect of industrial development prospects

Inactive Publication Date: 2017-06-16
南京佰泰克生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When HDACI directly acts on DCs, it reduces the ability of DCs to migrate to the chemokine CCL19 during the maturation process, thereby limiting the ability of DCs to enter lymphoid organs to perform antigen presentation functions; The activation of lymphocytes is crucial, and HDACI affects DC differentiation, maturation and migration, and interferes with its antigen presentation process

Method used

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  • Application of histone deacetylase and inhibitor thereof to preparation of tumor whole cell antigen loaded DC (dendritic cell) tumor vaccine
  • Application of histone deacetylase and inhibitor thereof to preparation of tumor whole cell antigen loaded DC (dendritic cell) tumor vaccine
  • Application of histone deacetylase and inhibitor thereof to preparation of tumor whole cell antigen loaded DC (dendritic cell) tumor vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: Effect of SAHA on Histone Deacetylase Activity and DC Vaccine Efficacy

[0025] 1. Experimental materials

[0026] C57BL / 6 mice were 8-week-old female SPF grade, purchased from the Animal Center of Nanjing University; the lung cancer cell line 3LL derived from C57BL / 6 mice was preserved by our company; DMEM medium and fetal bovine serum were purchased from Gibco.

[0027] All other reagents used were of domestic or imported analytical grade.

[0028] 2. Experimental method

[0029] 1. Isolation, preparation and identification of mouse bone marrow DC

[0030] The C57BL / 6 mice were sacrificed by neck dislocation, the femurs and tibias of both hind legs were removed, the attached muscle tissues were removed, both ends of the bones were removed, the bone marrow cavity was exposed, the bone marrow cavity was washed several times with PBS, and a single cell suspension was made by blowing thoroughly. After filtering with a 200-mesh copper mesh, wash twice with PB...

Embodiment 2

[0055] Example 2: Screening of histone deacetylase inhibitors

[0056] Screening model: C57BL / 6 mouse lung cancer cell line 3LL was cultured in DMEM medium containing 10% FBS, cells in the logarithmic growth phase were selected and adjusted to 1×10 7 / mL, grouped, respectively added the benzothiazole derivatives in the following table to the culture medium, after 6 hours of cultivation, replaced with the medium without benzothiazole derivatives and continued to cultivate for 6 hours, adjusted to 5×10 6 / mL, add 200μl of lysate (50mM pH 7.4 Tris-HCl, 150mM NaCl, 1% Triton X-100, 0.5mM EDTA, add PMSF just before use to make the final concentration 1mM), blow slowly for about 5min, collect the lysate in In an EP tube, shake on a decolorizing shaker at 4°C for 20 minutes, centrifuge at 12,000×g for 15 minutes at 4°C, collect the supernatant, filter through a 0.2 μm microporous membrane, and store at -80°C. The control group did not add benzothiazole derivatives, and other operati...

Embodiment 3

[0061] Example 3: Effects of Histone Deacetylase Inhibitors on the Efficacy of DC Vaccines

[0062] 1. Experimental materials

[0063] With embodiment 1.

[0064] 2. Experimental method

[0065] 1. Isolation, preparation and identification of mouse bone marrow DC

[0066] With embodiment 1.

[0067] 2. Lung cancer cell line 3LL was incubated with HDACs inhibitors and lysed

[0068] Experimental group: C57BL / 6 mouse lung cancer cell line 3LL was cultured in DMEM medium containing 10% FBS, cells in the logarithmic growth phase were selected and adjusted to 1×10 7 / mL, add the benzothiazole derivatives 1-3 (respectively experimental group 1-3) of structure and concentration in the embodiment 2 in culture fluid, after cultivating 6 hours, change to the culture medium that does not contain HDACs inhibitor Continue culturing for 6 hours and adjust to 5×10 6 / mL, add 200μl of lysate (50mM pH 7.4 Tris-HCl, 150mM NaCl, 1% Triton X-100, 0.5mM EDTA, add PMSF just before use to make...

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Abstract

The invention discloses the application of histone deacetylase and its inhibitor in the preparation of DC tumor vaccine loaded with tumor whole cell antigen. The applicant's previous research found that the tumor whole cell antigen treated with SAHA can significantly improve the preventive and therapeutic effects of DC tumor vaccines on tumors, and further studied the effect of SAHA on the target histone deacetylase and tumor whole cell antigen loaded DC tumor vaccine therapeutic relationship. The results prove that histone deacetylase is related to the curative effect of DC tumor vaccine loaded with tumor whole cell antigen, and the histone deacetylase inhibitor provided by the present invention can enhance the effect of tumor whole cell antigen on DC through its target histone deacetylase. The sensitization effect of DC can significantly improve the preventive and therapeutic effects of DC tumor vaccines on tumors. The target and inhibitor can be used to prepare DC tumor vaccines with higher preventive and therapeutic value.

Description

technical field [0001] The invention belongs to the field of tumor immunity and relates to tumor vaccines, in particular to the application of histone deacetylase and its inhibitors in the preparation of DC tumor vaccines loaded with tumor whole cell antigens. Background technique [0002] Immunotherapy is the fourth tumor treatment method after surgery, radiotherapy, and chemotherapy. In the comprehensive treatment mode of tumors, immunotherapy plays an important role in improving the cure rate of tumors, improving the quality of life of patients, and prolonging survival. In tumor immunity, tumor antigens recognized by T lymphocytes are not presented by tumor cells, but by antigen-presenting cells. Dendritic cells (DC) are the most powerful antigen-presenting cells known in the human body and the only antigen-presenting cells that can activate resting T cells, and have a strong ability to activate helper and cytotoxic T cells. Dendritic cell tumor vaccine is one of the fou...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61K31/428A61P35/00C12Q1/34
CPCA61K39/0011A61K31/428A61K2039/5154C12Q1/34
Inventor 胡攀勇张钟祥
Owner 南京佰泰克生物技术有限公司
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