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Use of recombinant mesenchymal stem cell in preparation of immunosuppressant

An immunosuppressant and mesenchymal stem cell technology, which is applied in the field of stem cell therapy, can solve the problems of increased tumor recurrence, weak immune regulation ability, and large amount of cells, and achieve the effects of suppressing immune response, reducing the risk of tumor recurrence, and reducing embolism

Active Publication Date: 2015-07-29
INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Based on the disadvantages of ordinary mesenchymal stem cells in clinical treatment, such as weak immunoregulatory ability in vivo, large amount of cells used, and the potential to increase the risk of tumor recurrence / occurrence, more economical and efficient cell therapy strategies that can overcome the above disadvantages are currently available. particularly urgent

Method used

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  • Use of recombinant mesenchymal stem cell in preparation of immunosuppressant
  • Use of recombinant mesenchymal stem cell in preparation of immunosuppressant
  • Use of recombinant mesenchymal stem cell in preparation of immunosuppressant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Preparation of recombinant mesenchymal stem cells expressing chemokine receptor CCR7

[0043] 1. Isolation and culture of mouse bone marrow MSCs

[0044] The sample is 2-3 weeks old C57BL / 6 mice (Experimental Animal Center, Academy of Military Medical Sciences), the mice were killed by neck dislocation, the mice were soaked in 75% alcohol to sterilize the mice → the mouse tibia and femur were separated under aseptic conditions and placed in a plate →Use sterile gauze to rub the attached muscle and remove the metaphysis →Rinse the bone marrow cavity repeatedly with PBS+10% FCS (Hyclone) to flush out the bone marrow cells →Inoculate in a culture bottle, so that the bone fragments are evenly distributed on the bottom of the bottle, and the culture system Containing αt-MEM (Gibco) + 10% FCS (Hyclone) + 100U / ml penicillin + 100U / ml streptomycin, 37°C, 5% CO 2 Cultivate under saturated humidity conditions → change the medium once every 3 days → when the cells are c...

Embodiment 2

[0059] Example 2 Detection of migration ability and enrichment area of ​​MSCs to secondary lymphoid organs in vivo

[0060] 1. Detection of migration ability of MSCs to secondary lymphoid organs in vivo

[0061] MSCs, MSCs / eGFP or MSCs / CCR7-eGFP (1×10 6 ) three groups of mice (4 mice in each group); 5 days after the infusion, the spleen (SP), lymph node (LN), mesenteric lymph node (MLN) and PP node were taken for frozen sections, and DAPI was used for nuclear labeling. Green fluorescent cells were observed under a fluorescence microscope. figure 2 A The results showed that MSCs / eGFP did not have the ability to migrate to secondary lymphoid organs, while MSCs / eGFP-CCR7 had a large distribution in SP, LN, MLN, and PP. figure 2 Calculate the eGFP of each organ as shown in B +The frequency of distribution of MSCs in various tissues and organs in vivo (the number of green fluorescence / high power field) quantitatively shows that the infused MSCs / eGFP does not have the ability t...

Embodiment 3

[0064] Example 3 Detection of MSCs / CCR7-eGFP immune regulation effect in vivo and its impact on tumor growth

[0065] 1. Inhibition of GvHD by MSCs / CCR7-eGFP

[0066] GvHD is a typical T cell-mediated immune disease. Establishment of mouse GvHD model: Take 20-24g BALB / c male mice, collect splenocytes and bone marrow cells, and count; 2×10 7 Splenocytes+1×10 7 Bone marrow cells / 0.25mlPBS were injected into the tail vein of 22-24g C57BL / 6 male mice irradiated with 9Gy to establish aGvHD model (GvHD group). On this basis, a total of 1×10 5 or 1×10 6 MSCs / eGFP, 1 x 10 5 or 1×10 6 MSCs / CCR7-eGFP. 1×10 6 Mice co-infused with MSCs / eGFP tail vein are called GvHD+MSCs / eGFP mice; 1×10 6 Mice with tail vein co-infusion of MSCs / CCR7-eGFP were called GvHD+MSCs / CCR7-eGFP mice.

[0067] After infusion, the survival and status of the mice in the three groups were observed daily, and the survival curves were recorded to observe the immune regulation effect of MSCs in vivo, and the le...

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Abstract

The invention discloses a use of a recombinant mesenchymal stem cell in preparation of an immunosuppressant. The invention relates to the recombinant mesenchymal stem cell capable of expressing a chemokine receptor CCR7 on a cytomembrane. After infusion, the recombinant mesenchymal stem cells can be specifically transferred to a secondary lymphatic organ, and a large amount of the recombinant mesenchymal stem cells are gathered in a T cell enrichment region in the secondary lymphatic organ and can efficiently inhibit T cell immunoreaction. Through use of the recombinant mesenchymal stem cell in treatment on graft-versus-host disease after bone marrow transplantation, immunological rejection after organ transplantation and autoimmune diseases, a cost is reduced, a cell use amount is reduced and side-effects are relieved. In treatment, the recombinant mesenchymal stem cell retains tumor cell killing effects and provides a high efficiency and beneficial novel approach for clinical treatment on graft-versus-host disease after bone marrow transplantation, immunological rejection after organ transplantation and autoimmune diseases of tumor high-risk groups.

Description

technical field [0001] The invention relates to the field of stem cell therapy, in particular to the application of a mesenchymal stem cell whose cell membrane expresses chemokine receptor CCR7 in the preparation of an immunosuppressant. Background technique [0002] Mesenchymal stem cells are the first group of adult stem cells isolated from bone marrow with self-renewal and multilineage differentiation potential. Recent studies have shown that mesenchymal stem cells have a wide range of immune regulation functions. Mesenchymal stem cells have immunoregulatory effects on a variety of immune cells, including T cells, B cells, NK cells, and DCs. In effective animal experiments and treatment of clinical diseases, it has been found that mesenchymal stem cells play an immunoregulatory role in vivo mainly through secreted cytokines. [0003] At present, the immunotherapy of mesenchymal stem cells on clinical diseases such as hematopoietic stem cell transplantation or graft-vers...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61P37/06C12N5/10C12N15/87C12N15/63
Inventor 李红郭希民毛宁江小霞
Owner INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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