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Composition for relieving immunosuppression of immune cells, antigen-presenting cells and preparation method thereof

A technology of antigens and cells, applied in the fields of immunology and medicine

Active Publication Date: 2021-02-23
启辰生生物科技(珠海)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are currently no drugs related to Gal9-binding peptide targets that have been approved for marketing at home and abroad.

Method used

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  • Composition for relieving immunosuppression of immune cells, antigen-presenting cells and preparation method thereof
  • Composition for relieving immunosuppression of immune cells, antigen-presenting cells and preparation method thereof
  • Composition for relieving immunosuppression of immune cells, antigen-presenting cells and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0052] [Preparation method of engineered antigen-presenting cells]

[0053] The third aspect of the present invention provides a method for preparing engineered antigen-presenting cells, the engineered antigen-presenting cells are used to relieve immune cell immunosuppression, which includes introducing the composition described in the first aspect into the antigen-presenting cells , so as to engineer antigen-presenting cells.

[0054] In certain embodiments, the methods of the invention comprise the steps of:

[0055] (1) constructing nucleic acids capable of producing components (a) and (b);

[0056] (2) isolate peripheral blood mononuclear cells from venous blood, and induce differentiation to obtain antigen-presenting cells; and

[0057] (3) introducing the nucleic acid of step (1) into the antigen-presenting cell of step (2), and culturing the antigen-presenting cell under conditions suitable for expression of the nucleic acid.

[0058] In a preferred embodiment, the m...

preparation example

[0062] This preparation example is to prepare DNA and mRNA encoding antigens and composition components

[0063] 1. Preparation of DNA and mRNA Constructs

[0064] The DNA sequences used to produce the mRNA encoding the TGF-β regulatory peptide and the PD1-Gal9 binding peptide of the present invention were respectively constructed, and used for subsequent in vitro transcription reactions. Following the coding sequence is a polyadenosine segment. The DNA sequence information is shown in Table 1 below.

[0065]In addition, the coding sequence of human tumor antigen GPC3 for in vitro sensitization was constructed. The sequence of GPC3 is available through the Genebank database. In this example, the antigen disclosed in CN107583042A was used.

[0066] Table-1 DNA sequence list

[0067]

[0068] 2. In vitro transcription

[0069] Firstly, the prepared corresponding DNA plasmid was linearized using restriction endonuclease, and the linearized plasmid was used as a template,...

Embodiment 1

[0071] This example was used to study the effect of exemplary compositions of the invention on T cell responses.

[0072] 1. Induction culture of DC cells in vitro

[0073] Aseptically extract 50ml of venous blood from a healthy person, separate peripheral blood mononuclear cells with lymphocyte separation medium in an ultra-clean workbench, add the mononuclear cells to the AIM-V medium, and place them in 37°C, 5% CO 2 Incubate in an incubator to allow monocytes to adhere to the wall. After 2h, the non-adherent cells were removed, and the adherent cells were added to iDC medium (GM-CSF with a final concentration of 800U / mL and IL-4 at 500U / mL were added to the AIM-V medium), and placed at 37°C for 5 %CO 2 Cultured in the incubator for 6 days. Transfer half of the cell culture medium to a centrifuge tube, collect the cells by centrifugation at 500g, remove the supernatant, and add an equal volume of fresh mDC medium (configuration of fresh medium for mDC: 1600U / mL GM-CSF and...

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Abstract

The invention discloses a composition for releasing immunosuppression of immune cells, an antigen-presenting cell and a preparation method thereof. The composition of the present invention comprises the following (a) and (b) components or nucleic acids encoding them: (a) soluble TGF-β regulatory peptides, mutants or functional fragments thereof and (b) soluble PD1 and soluble Gal9 binding peptides Fusion protein, mutants or functional fragments thereof. The composition of the present invention can synergistically improve the immune response of T lymphocytes.

Description

technical field [0001] The invention relates to the fields of immunology and medicine, in particular to a composition for releasing immune cell immunosuppression, an antigen-presenting cell and a preparation method thereof. Background technique [0002] In recent years, there have been numerous reports of the use of antigen-loaded DC vaccines in the treatment of tumors, and from the currently reported data, DC vaccines appear to represent a new and very promising approach for improved tumor immunotherapy. However, the use of DC vaccine alone usually does not lead to the expected improvement of the immunotherapy effect, and a satisfactory clinical effect cannot be obtained. [0003] PD-1 is expressed on activated T cells, B cells, and on monocytes involved in mediating the balance between immune activation and tolerance. Its main role is to limit T cell activity in the inflammatory environment and to limit autoimmunity in the inflammatory response to infection. The basis of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/18A61K39/395A61P35/00C12N5/10C12N15/87
CPCA61K38/1841A61K39/39566A61P35/00C12N5/0639C12N15/87C12N2510/00A61K2300/00
Inventor 蒋俊林鑫
Owner 启辰生生物科技(珠海)有限公司
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