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CD33 immunogenic polypeptide and application thereof

An immunogen and cellular immunity technology, applied in the direction of specific peptides, cancer antigen components, vertebrate antigen components, etc., can solve the problems of specific T cell immunity, large molecular weight of CD33, difficult CD33, etc., to promote T cell proliferation, Effects of inhibiting growth and inhibiting CD33 expression

Inactive Publication Date: 2017-01-18
SUZHOU PULUODA BIOLOGICAL SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, CD33 has a large molecular weight, and it is difficult to obtain high-purity CD33
In this way, it will not only bring difficulties to the specific T cell immunity in the later stage, but also lead to the patient's autoimmunity

Method used

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  • CD33 immunogenic polypeptide and application thereof
  • CD33 immunogenic polypeptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] The in vivo activity of CD33 immunogen polypeptide was detected by acute myeloid leukemia mouse model.

[0015] SCID mice aged 6-8w were randomly divided into 4 groups, half male and half male, 10 mice in each group. (1) blank group; (2) low-dose polypeptide group; (3) medium-dose polypeptide group; (4) high-dose polypeptide group. The leukemia animal model was established by intraperitoneal injection of HL-60 cells, and the acute myeloid leukemia mouse model was established. Immunization was carried out on the 3rd, 5th, and 7th day after inoculation of tumor cells. The scheme is: add the same volume of solvent to the blank group, set three doses of polypeptide in the experimental group: 2, 5, and 10 mg / Kg, and inject at multiple points around the tumor. After 21 days, the number of surviving mice was observed, and the survival rate was calculated. The results showed that the peptides at doses of 2, 5, and 10 mg / Kg could effectively protect tumor-bearing mice and impr...

Embodiment 2

[0017] Use the competitive receptor-ligand affinity method to test the binding ability of peptides to MHC:

[0018] The ability of the polypeptide to bind to each type of HLA was judged by the competitive receptor-ligand affinity method. Add 125I-labeled polypeptides with a fixed concentration of 50 μmol / L and polypeptides with different concentrations (1-50 μmol / L) to the reaction system (phosphate buffer and MHC, the MHC kit was purchased from Shanghai Fanke Biotechnology Co., Ltd. , there are HLA-A1, A2, A3, A11 and A24 in the kit), and two complexes are formed in the reaction system, that is, the polypeptide MHC complex to be tested and the 125I-labeled polypeptide complex. The polypeptide to be tested competes with the 125I-labeled polypeptide for binding to MHC. Use ultrafiltration (product model Microcon 30, Amicon company) to separate free polypeptide and polypeptide MHC complex, then measure the 125I radioactivity of the polypeptide MHC complex, and compare this radi...

Embodiment 3

[0020] Proliferation of T lymphocytes: Spleen of mice was aseptically taken, washed 3 times with 1640 medium, ground with a 5ml syringe core, filtered with a 200-mesh screen to make a single-cell suspension, centrifuged (1000r / min, 5min), discarded Qing, Tris-NH 4 CL cracked red blood cells, put them in an ice-water bath for 3-5min, centrifuged (1000r / min, 5min), discarded the supernatant, and washed the cells twice with sterile cold PBS. Add RPMI 1640 culture fluid (5ml) of 10% calf serum to suspend the cells at last, count the cells, and adjust the cell concentration to be 5×10 6 cells / ml and cultured in 96-well culture plates.

[0021] The experiment set up a blank control group, a model group (concanavalin A, purchased from sigma company), and groups of different doses of polypeptides (10, 20, 40 mg / ml). After adding 100 μl / well of spleen lymphocyte suspension to each group, 100 μl of 1640 culture medium was added to the blank control group, ConA (final concentration was...

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Abstract

The invention discloses a CD33 immunogenic polypeptide, belongs to the field of anti-cancer vaccines, and particularly relates to a CD33 immunogenic optimized polypeptide which serves as an anti-cancer vaccine to enhance T cell immune response. The amino acid sequence of the CD33 immunogenic optimized polypeptide is a brand new sequence, and the CD33 immunogenic optimized polypeptide is applied to preparation of drugs used for treating tumors, especially applied to preparation of drugs used for being applied to tumor immune cells and enhancing the T cell immune response and applied to preparation of drugs used for CAR-T cellular immunotherapy. The CD33 immunogenic polypeptide has the advantages that the amino acid sequence is the brand new sequence, and the CD33 immunogenic polypeptide can serve as an immunogen in a tumor immune cell technology, promote T cell proliferation, inhibit expression of CD33 on the surfaces of peripheral blood leukocytes of mice, be effectively and specifically combined with MHC on the surfaces of T cells, inhibit tumor growth in a body of an acute myelogenous leukemia mouse model and increase the survival rate. The CD33 immunogenic polypeptide is applied to cell therapy of CAR-T and the like by serving as a target molecule of the cell therapy.

Description

technical field [0001] The present invention relates to the field of anticancer vaccines. In particular, the present invention relates to optimized polypeptides for use as CD33 immunogens in anticancer vaccines to enhance T cell immune responses. Background technique [0002] Tumor cell immunotherapy is an emerging tumor treatment model with significant curative effect, and it is a new treatment method for autoimmune anti-cancer. It is a method of using biotechnology and biological agents to culture and expand immune cells collected from patients in vitro, and then reinfuse them back into the patient's body to stimulate and enhance the body's autoimmune function, so as to achieve the purpose of treating tumors. Tumor cell immunotherapy is the fourth largest tumor treatment technology after surgery, radiotherapy and chemotherapy. [0003] CD33 is a myeloid cell differentiation antigen with a molecular weight of 67KD, which is mainly distributed in myeloid blood cells, espec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/705A61K39/00A61K35/17A61P35/00
CPCC07K14/705A61K35/17A61K39/0011
Inventor 罗瑞雪
Owner SUZHOU PULUODA BIOLOGICAL SCI & TECH
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