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Transmembrane heparan sulfate proteoglycan 1 immune agonist polypeptide and application thereof

A technology of acetylheparin sulfate and agonist, which is applied in the direction of animal/human protein, specific peptide, vertebrate antigen components, etc., can solve the problems of difficult Syndecan-1, specific T cell immunity, large molecular weight, etc., and achieve Improve the survival rate, promote the proliferation of T cells, and promote the effect of binding

Inactive Publication Date: 2017-02-01
SUZHOU PULUODA BIOLOGICAL SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, syndecan-1 molecules are divided into extracellular segment, transmembrane segment and cytoplasmic segment, and the molecular weight is relatively large, so it is difficult to obtain high purity Syndecan-1
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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  • Transmembrane heparan sulfate proteoglycan 1 immune agonist polypeptide and application thereof
  • Transmembrane heparan sulfate proteoglycan 1 immune agonist polypeptide and application thereof
  • Transmembrane heparan sulfate proteoglycan 1 immune agonist polypeptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] The in vivo activity of Syndecan-1 immune agonist polypeptide was detected by tumor model.

[0016] C57BL / 6 mice aged 6-8 weeks 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 non-small cell lung cancer tumor model was established, and 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, and set three doses of polypeptides in the experimental group: 5, 10, and 20 mg / Kg, and inject them at multiple points around the tumor. After 21 days, the number of surviving mice was observed, and the survival rate was calculated. The results show that the polypeptide can effectively protect mice and improve the survival rate of tumor-bearing mice, and the survival rate of the dose 20mg / Kg group reaches 53.3%.

Embodiment 2

[0018] 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.

[0019] The experiment set up a blank control group, a model group (concanavalin A, purchased from sigma company), and groups of different doses of polypeptides (5, 10, 20 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 ...

Embodiment 3

[0025] The effect of Syndecan-1 immune agonist polypeptide on CD8+ T cells was determined by flow cytometry. C57BL / 6 mice aged 6-8 weeks were randomly divided into 4 groups, half male and half male, 10 mice in each group. (1) blank group; (2) low-dose immune agonist polypeptide group; (3) medium-dose immune agonist polypeptide group; (4) high-dose polypeptide group. On the 0th, 7th, and 14th day of the experiment, immunization was carried out respectively. The scheme is: add the same volume of solvent to the blank group, and set three doses of polypeptides in the experimental group: 5, 10, and 20 mg / Kg, and inject them at multiple points near the lymph nodes on the back of the mice. After 30 days, 0.8 ml of blood was taken from the eyeball, anticoagulated with heparin, and the blood cell layer was collected after centrifugation, and the red blood cells were cracked with red blood cell lysate, and the cracked red blood cells were washed away, and then fluorescently labeled mon...

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Abstract

The invention discloses a transmembrane heparan sulfate proteoglycan 1 immune agonist polypeptide, belongs to the field of anti-cancer vaccines, and particularly relates to an optimized transmembrane heparan sulfate proteoglycan 1 immune agonist polypeptide used as an anti-cancer vaccine to enhance T cell immune responses. The amino acid sequence is a brand new sequence. The polypeptide is applied to preparing medicine for treating tumors, particularly to preparing medicine for tumor immune cells and T cell immune response enhancing and preparing medicine for CAR-T cell immune treatment. The polypeptide has the advantages that the sequence is brand new, the polypeptide can be used in an immune agonist in the tumor immune cell technology, promote T cell proliferation, promote CD8+ immune response, promote combination of T cells and tumor cells and increase the survival rate in mice bearing neoplasm, and serves as a target molecule of cell therapies to be applied to CAR-T and other cell therapies.

Description

technical field [0001] The present invention relates to the field of anticancer vaccines. Specifically, the present invention relates to an optimized polypeptide used as a transmembrane heparin sulfate proteoglycan 1 immune stimulant for an anticancer vaccine to enhance T cell immune response. Background technique [0002] Transmembrane heparan sulfate proteoglycan 1 (Syndecan-1), a member of the adhesion molecule integrin transmembrane adhesion protein (heparan sulfate proteoglycan, HSPG) family, can bind to a variety of factors, participate in the differentiation and development of tissues and organs, blood vessels It regulates a series of physiological processes such as formation and tissue regeneration, and is related to processes such as tumor cell homing and metastasis, and is also an indicator for judging the prognosis of certain tumors. [0003] The study found that, except for the most superficial terminally differentiated cells in a normal human body, almost all h...

Claims

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Application Information

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IPC IPC(8): C07K14/705A61K39/00A61K35/17A61P35/00
CPCA61K35/17A61K39/0011C07K14/705A61K2300/00
Inventor 罗瑞雪
Owner SUZHOU PULUODA BIOLOGICAL SCI & TECH
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