Application of B cell vaccine in preparation of drugs for treating rheumatoid arthritis

A B-cell and arthritis technology, applied in the field of B-cell vaccines, can solve problems such as limited selection, unsatisfactory vaccine treatment effect, and inability to cure, and achieve low frequency of administration, reduce bone erosion, and have no immune tolerance Effect

Active Publication Date: 2019-12-20
PEOPLES HOSPITAL PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current treatment plan still has certain limitations: lifelong medication is required, and most patients can only maintain remission but cannot be cured. Therefore, more specific, effective and simple treatment plans are urgently needed
[0008] Although there are therapeutic vaccines against RA, the clinical options are still limited, and most of the vaccines are not satisfactory in terms of therapeutic effect

Method used

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  • Application of B cell vaccine in preparation of drugs for treating rheumatoid arthritis
  • Application of B cell vaccine in preparation of drugs for treating rheumatoid arthritis
  • Application of B cell vaccine in preparation of drugs for treating rheumatoid arthritis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1CD19

[0034] Example 1CD19 + Preparation of total B cell vaccine

[0035] Experimental animals: Specific pathogen free (SPF) DBA / 1 male mice of 6-8 weeks purchased from Beijing Huafukang Biotechnology Co., Ltd., all of which were fostered in the SPF-level environment of the Animal Laboratory of Peking University People's Hospital. Modeling experiment was carried out after 1 week of adaptive feeding.

[0036] Modeling method: Bovine Type II Collagen (C II) was dissolved in 100 mmol / L glacial acetic acid overnight at 4°C, and mixed with complete Freund's adjuvant (CFA) at a volume ratio of 1:1. Fully emulsified. Inject subcutaneously at two points at the base of the tail of the mouse, each injecting an emulsion containing 100 μg of C II; on day 21 after the initial immunization, C II and incomplete Freund's adjuvant (IFA) were injected at a volume ratio of 1:1 Fully emulsify, and subcutaneously inject the emulsion containing 100 μg CII at the base of the tail of the mouse to comple...

Embodiment 2

[0052] The preparation of embodiment 2Non-B10 B cell vaccine

[0053] (1) According to the methods (1) to (6) in Example 1, spleen cell pellets were obtained.

[0054] (2) Add 300 μL of normal saline to resuspend the spleen cell pellet, add 5 μL of surface antibodies CD19-FITC, CD5-APC, and CD1d-PE in the dark, vortex and shake at room temperature for 30 minutes in the dark (vortex once every 15 minutes).

[0055] (3) Add 2 mL of normal saline solution, pipette and mix well, then centrifuge at 1800 rpm for 5 min and discard the supernatant.

[0056] (4) Add 1mL of sorting solution (PBS containing 2% FBS) to resuspend the cell pellet, and then run it on a flow cytometer (FACS AriaII sorter from BD Company), and sort out B10 cells (CD19 + CD5 + CD1d hi ) except all B cells (CD19 + CD5 - / + CD1d low ), that is, to obtain Non-B10 B cells. Each CIA mouse can be sorted 1.5-2.0*10 7 A Non-B10 B cell (reference: Aregulatory B cell subset with a unique CD1d hi CD5 + phenotype ...

Embodiment 3

[0059] The preparation of embodiment 3Plasma cell vaccine

[0060] (1) According to the methods (1) to (6) in Example 1, spleen cell pellets were obtained.

[0061] (2) Add 300 μL of normal saline to resuspend the spleen cell pellet, then add 5 μL of surface antibodies CD19-FITC and CD138-APC in the dark. Just add enough antibody to fully bind the cells to the antibody, and add 5-10 μL according to the number of cells in each tube.

[0062] (3) Using flow cytometry, the phenotype of the sorted cells is CD19 - / + CD138 + Plasma cells.

[0063] (4) Then adopt the same treatment mode as steps (9) to (14) in Example 1 to prepare Plasma cell vaccine. The concentration is 1×10 5 cell / 300μL.

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Abstract

The present invention discloses an application of a B cell vaccine in preparation of drugs for treating rheumatoid arthritis and the B cell vaccine having the application. Compared with drug treatment, the B cell vaccine has the following advantages of being smaller in dosage, lower in administration frequency, longer in maintenance period, and simple and convenient in subcutaneous injection, cantreat diseases through autoimmune regulation functions, can effectively control the diseases, reduces bone erosion and joint destruction, and is free of immune tolerance problems in autotransfusion and small in side effects.

Description

technical field [0001] The invention relates to the technical field of drugs for treating autoimmune diseases, in particular to the application of B cell vaccine in treating rheumatoid arthritis and the B cell vaccine with the application. Background technique [0002] Rheumatoid arthritis (RA) is a disease of irreversible recovery of bone erosion with a high rate of disability. However, the current treatment plan still has certain limitations: lifelong medication is required, and most patients can only maintain remission but cannot be cured. Therefore, a more specific, effective and simple treatment plan is urgently needed. [0003] At present, the most commonly used drugs for RA treatment are mainly divided into c-DMARDs and b-DMARDs; in addition, they also include newly developed small molecule drugs, such as JAK inhibitors and BTK inhibitors. [0004] The pathogenesis of RA is closely related to the excessive activation of B cells to produce a large number of autoantibo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61P29/00A61P19/02
CPCA61K39/0008A61P29/00A61P19/02C12N5/0635C12N2509/00A61K2039/515
Inventor 栗占国胡凡磊宋婧
Owner PEOPLES HOSPITAL PEKING UNIV
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