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Composition, immune cell and application for enhancing T-lymphocyte immunity

A technology of immune cells and compositions, applied in the field of biomedicine, can solve the problems of low response rate, inability to achieve ideal effects, and improvement of immunotherapy effects.

Active Publication Date: 2020-09-11
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

It shows that the use of IL12 alone cannot achieve the desired effect[Mazzolini G D, Alfaro C, Sangro B, et al.1134. Official Journal of the American Society of Clinical Oncology,2005,23(5):999-1010.]
[0009] Current clinical trials have shown that the response rate of DC therapeutic vaccines seldom exceeds 15%, and the overall response rate is low. The use of DC vaccine alone usually cannot lead to the expected improvement in the effect of immunotherapy, and cannot obtain satisfactory results. clinical effect

Method used

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  • Composition, immune cell and application for enhancing T-lymphocyte immunity
  • Composition, immune cell and application for enhancing T-lymphocyte immunity

Examples

Experimental program
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Embodiment 1

[0067] This example is for the preparation of DNA and mRNA encoding antigens and immune checkpoint inhibitors

[0068] 1. Preparation of DNA and mRNA Constructs

[0069] DNA sequences encoding IL12, IL15, IL15Rα, and PD1-Fc-CD80 mRNA were respectively constructed for this example, and used for subsequent in vitro transcription reactions. After the coding sequence, there is a polyadenosine fragment to prepare the construct, and the specific sequence is shown in Table 1.

[0070] In addition, the coding sequence of human tumor antigen GPC3 for in vitro sensitization is constructed. The coding sequence of GPC3 in the present invention consists of the sequence shown in SEQ ID No.8, and the amino acid sequence consists of the sequence shown in SEQ ID No.9. The sequence of GPC3 is available through the Genebank database. In this example, the antigen disclosed in CN107583042A was used.

[0071] Table-1 Nucleic acid sequence list

[0072] name nucleic acid sequence num...

Embodiment 2

[0076] This example was used to study the effect of immunomodulator compositions on T cell responses.

[0077] 1. Induction culture of DC cells in vitro

[0078] Aseptically extract 50ml of venous blood from patients with hepatocellular carcinoma, separate peripheral blood mononuclear cells with lymphocyte separation medium in an ultra-clean workbench, add mononuclear cells to 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: add AIM-V medium...

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Abstract

The invention discloses a composition and immune cells for enhancing T lymphocyte immunity and application. The composition of the present invention comprises the following components (a) to (d): (a)interleukin-15, a mutant thereof or a functional fragment thereof, or an encoding nucleic acid thereof; (b) interleukin-15 receptor alpha, a mutant or a functional fragment thereof, or an encoding nucleic acid thereof; (c) interleukin-12, a mutant or a functional fragment thereof, or an encoding nucleic acid thereof; and (d) a PD1-CD80 fusion protein, a mutant or a functional fragment thereof, oran encoding nucleic acid thereof. The composition, the immune cells and the method for improving the properties of the T lymphocytes can all excite stronger immune response, and induce tumor specificCD4 and CD8T cells in a higher proportion, so that the defects of tumor immunosuppression and non-ideal clinical effect which are difficult to solve by a single tumor treatment means are expected to be overcome.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a composition for enhancing the immunity of T lymphocytes, immune cells and applications. Background technique [0002] The body's immune response is first captured by antigen-presenting cells (APCs), and after processing and processing, the antigen information is presented to lymphocytes, and then a series of specific immune responses are triggered. Dendritic cells (DC) are considered to be the most functional APCs at present, and their biggest feature is that they can stimulate the proliferation and activation of Naive T cells, and are the center of initiating, regulating and maintaining specific immune responses links. In the body's anti-tumor immunity, cellular immunity mediated by T cells plays an important role. [0003] Studies have found that tumor patients have the characteristics of reduced number of DCs and functional defects. The number and function of infiltrating DCs in...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/39A61K39/00A61P35/00
CPCA61K39/0011A61K39/39A61K2039/5154A61K2039/55516A61K2039/55527A61K2039/55538A61K2039/585A61P35/00
Inventor 蒋俊张欣月文高柳
Owner 启辰生生物科技(珠海)有限公司
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