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A kind of immunosuppressive drug and preparation method thereof

An immunosuppressive and drug technology, applied in the field of immunosuppressive drugs and their preparation, can solve problems such as immune hyperactive diseases, reduce the chance of cross-infection or iatrogenic infection, facilitate long-term storage, and reduce the number of blood collections Effect

Active Publication Date: 2022-02-01
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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  • A kind of immunosuppressive drug and preparation method thereof
  • A kind of immunosuppressive drug and preparation method thereof
  • A kind of immunosuppressive drug and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] The present invention is the preparation of a medicament immunosuppressive

[0052] 1. Experimental Materials and Reagents

[0053] Experiments with C57BL / 6 mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., 6-8 weeks of age, weighing approximately 18g.

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

[0055] RPMI-1640 medium: commercially available from the company PAN;

[0056] Mouse pancreatic stromal cell line Y0501 formation object oriented storage system;

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

[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 antibody was purchased from Biolegend Company.

[0059] 2. Experimental Procedure

[0060] (1) mice were isolated with Ficoll solution (density 1.083) peripheral blood mononuclear cells;

[0061] (2) the isolated peripheral blood mononu...

Embodiment 2

[0072] Regulatory dendritic cells adoptive reinfusion treatment of rheumatoid arthritis

[0073] 1. Experimental Materials and Reagents

[0074] Experiments with DBA / 1 mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., 6-8 weeks of age, weighing approximately 18g.

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

[0076] 2. Experimental Procedure

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

[0078] (2) Day 0 type II collagen emulsified injected into mice tail root receptor 100uL, day 42 and day 21, recipient mice were injected at the same site of the same dose;

[0079] (3) Day 22, the recipient mice were divided into two groups, one group was injected with 1 × 10 6 Regulatory dendritic cells (prepared as in Example 1), another group was injected with PBS;

[0080] (4) the degree of swelling was observed on day 50 i...

Embodiment 3

[0084] Regulatory dendritic cells overlay return to the treatment of diabetic models Rip-ova mice

[0085] Experimental materials and reagents

[0086] Experiments were purchased from Beijing Virong Lihua Experimental Animal Technology Co., Ltd., aged 6-8 weeks, with a weight of 18g; Rip-Ova mice and OT-I mouse were purchased from Jackson laboratory, age 6 -8 weeks, weighing around 18g.

[0087] OVA 257-264 Polypeptide Buy Self-China CHINESE PEPTIDE COMPANY (China Hangzhou);

[0088] Anti-mouse CD8A magnetic bead antibody purchases from German Mei Tianzhu Company;

[0089] Blood glucose detectors purchase self Touch branded from Johnson.

[0090] 2. Experimental steps

[0091] (1) Separate OT-I mouse spleen cells, using a Japanese 旎 Anti-Mouse CD8A magnetic bead antibody to isolate OT-I mouse CD8T cells; take 1 × 10 6 OT-I mouse CD8T cells, add 1 × 10 5 Ripe tree prototype (cultured in Example 1), and add 1 ug / ml concentration of OVA in a 10% FCS-containing RPMI-1640 medium 257-...

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Abstract

The invention provides an immunosuppressive drug and a preparation method thereof. The immunosuppressive drug comprises dendritic cells induced by pancreatic stromal cells and having a negative immune regulation effect. The present invention also provides a method for preparing the immunosuppressive drug, comprising 1) obtaining mononuclear cells from mammalian blood and inducing them into mature dendritic cells; 2) co-culture of pancreatic stromal cells and dendritic cells , inducing regulatory dendritic cells; 3) collecting regulatory dendritic cells that play a role in negative immune regulation. The immunosuppressive drug provided by the invention has good immunosuppressive effect, can be used for treating rheumatoid arthritis and type I diabetes, and has good use safety.

Description

Technical field [0001] The present 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 the central link of an immune response. After the foreign antigen enters the body, dendritic cells can be identified, processed, treated, and delivered to CD4T cells and CD8T cells to excite the latter immune response. In 2004, a group of dendritic cell subgroups with immune negative adjustment energy (1) were reported. The subpopulation low expression MHC-II molecules, low expression CD40, CD80, CD86, etc. Therefore, it is named to regulate dendritic cells. Further studies have found that regulatory dendritic cells can exert an immunosuppressive effect by secreting NO. [0003] Immunoharmal disease is due to excessive activation of the immune system, resulting in diseases that cause attack and destruction of self-antigens, including rheumat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0784A61K35/15A61P37/06A61P3/10A61P29/00A61P19/02
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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