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Immunosuppressive drug and preparation method thereof

A technology of immunosuppression and drugs, which is applied in the field of immunosuppressive drugs and its preparation, can solve the problems of no immune hypersensitivity diseases, reduce the chance of cross-infection or iatrogenic infection, suppress immune response, and reduce medical operations Effect

Active Publication Date: 2018-12-21
BEIJING ZHONGTAI HENGJI BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most of the treatment methods for immune hypersensitivity diseases are hormone therapy, and the application of immunotherapeutic methods such as IL-6 antibody and mesenchymal stem cells has begun, but there is no patent for the application of regulatory dendritic cells to treat immune hypersensitivity diseases

Method used

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  • Immunosuppressive drug and preparation method thereof
  • Immunosuppressive drug and preparation method thereof
  • Immunosuppressive drug and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Preparation of immunosuppressive drug of the present invention

[0052] 1. Experimental Materials and Reagents

[0053] The C57BL / 6 mice used in the experiment were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., aged 6-8 weeks, weighing about 18g.

[0054] Fetal calf serum (FCS): purchased from PAN company;

[0055] RPMI-1640 medium: purchased from PAN company;

[0056] The mouse pancreatic stromal cell line Y0501 was preserved for the establishment of this subject;

[0057] Mouse GM-CSF and IL-4 were purchased from Peprotech, USA;

[0058] FITC-labeled anti-mouse MHC-II, FITC-labeled anti-mouse CD40, FITC-labeled anti-mouse CD80, FITC-labeled anti-mouse CD86 and FITC-labeled isotype control antibodies were purchased from Biolegend, USA.

[0059] 2. Experimental steps

[0060] (1) Separation of mouse peripheral blood mononuclear cells with Ficoll separation medium (density 1.083);

[0061] (2) The isolated mouse peripheral blood mon...

Embodiment 2

[0072] Adoptive reinfusion of regulatory dendritic cells in rheumatoid arthritis

[0073] 1. Experimental Materials and Reagents

[0074] Experimental DBA / 1 mice were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., aged 6-8 weeks, weighing about 18 g.

[0075] Chicken type II collagen and complete Freund's adjuvant were purchased from Sigma-Aldrich, USA;

[0076] 2. Experimental steps

[0077] (1) Chicken type II collagen was emulsified in complete Freund's adjuvant to a final concentration of 1 mg / mL;

[0078] (2) On the 0th day, inject 100uL of emulsified type II collagen into the base of the tail of the recipient mice, and inject the recipient mice at the same site at the same dose on the 21st and 42nd days;

[0079] (3) On the 22nd day, the recipient mice were divided into two groups, and one group was injected with 1×10 6 Regulatory dendritic cells (preparation method is the same as in Example 1), and another group is injected with PBS; ...

Embodiment 3

[0084] Adoptive reinfusion of regulatory dendritic cells to treat diabetic model RIP-OVA mice

[0085] 1. Experimental Materials and Reagents

[0086] Experimental C57BL / 6 mice were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., aged 6-8 weeks, weighing about 18g; RIP-OVA mice and OT-I mice were purchased from Jackson Laboratories in the United States, aged 6 -8 weeks, weight about 18g.

[0087] OVA 257-264 Peptides were purchased from Chinese Peptide Company (Hangzhou, China);

[0088] Anti-mouse CD8a magnetic bead antibody was purchased from Miltenyi, Germany;

[0089] Glucose monitors were purchased from Johnson & Johnson under the One Touch brand.

[0090] 2. Experimental steps

[0091] (1) Isolate spleen cells from OT-I mice, and use anti-mouse CD8a magnetic bead antibody from Miltenyi Company to isolate CD8 T cells from OT-I mice; take 1×10 6 OT-I mouse CD8 T cells, add 1×10 5 Mature dendritic cells (cultivated according to the meth...

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Abstract

The invention provides an immunosuppressive drug and a preparation method thereof. The immunosuppressive drug comprises dendritic cells with an immunity negative regulating effect induced by pancreasstroma cells. The invention further provides a method for preparing the immunosuppressive drug. The method comprises the following steps: 1) to obtain a mononuclear cell from blood of a mammal, and inducing the mononuclear cell as a mature dendritic cell; 2) co-culturing the pancreas stroma cells and the dendritic cells, and inducing regulatory dendritic cells; and 3) collecting the regulatory dendritic cells for playing the immunity negative regulating effect. The immunosuppressive drug provided by the invention has the good immunosuppressive action, can be used for treating rheumatoid arthritis and diabetes type I, and has good using safety.

Description

technical field [0001] The invention relates to an immunomodulatory drug and a preparation method thereof, in particular to an immunosuppressive drug and a preparation method thereof. Background technique [0002] Dendritic cells are central to the immune response. After foreign antigens enter the body, dendritic cells can recognize, process, and process them, and present them to CD4 T cells and CD8 T cells, stimulating the latter's immune response. In 2004, a subpopulation of dendritic cells with the ability to negatively regulate immunity was reported (1). This subgroup has low expression of MHC-II molecules and low expression of co-stimulatory molecules such as CD40, CD80, and CD86; co-culture with CD4 T cells and CD8 T cells can significantly inhibit the ability of mature DC to expand T cells and exert a negative immune regulatory effect , hence the name regulatory dendritic cells. Further studies have found that regulatory dendritic cells can exert an immunosuppressi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/15A61P37/06A61P3/10A61P29/00A61P19/02C12N5/0784
CPCA61P3/10A61P19/02A61P29/00A61P37/06A61K35/15C12N5/0639C12N2502/22
Inventor 李政媛王超
Owner BEIJING ZHONGTAI HENGJI BIOLOGICAL TECH CO LTD
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