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B and T lymphocyte attenuator immunogen polypeptide and application thereof

A technology of weakening factor and lymphocytes, which is applied in the field of anti-cancer vaccines, can solve the problems of large molecular weight, difficult B/T lymphocyte weakening factor, and specific T cell immunity difficulties, so as to promote specific combination and T cell proliferation , the effect of inhibiting tumor growth

Inactive Publication Date: 2016-09-28
SUZHOU PULUODA BIOLOGICAL SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the B / T lymphocyte weakening factor is composed of a protein with 414 amino acids, and the molecular weight is relatively large, so it is difficult to obtain a high-purity B / T lymphocyte weakening factor
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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  • B and T lymphocyte attenuator immunogen polypeptide and application thereof
  • B and T lymphocyte attenuator immunogen polypeptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Human diffuse large B-cell lymphoma xenograft model was used to detect the activity of B / T lymphocyte weakening factor immunogen polypeptide in vivo.

[0016] 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. Human diffuse large B-cell lymphoma (DLBCL) xenograft model was established, and immunization was carried out on the 3rd, 5th, and 7th day after inoculation of DLBCL cells. The scheme is: add the same volume of solvent to the blank group, set three doses of polypeptides in the experimental group: 0.5, 1.0, and 2.0 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 can improve the survival rate of tumor-bearing mice at do...

Embodiment 2

[0018] The in vivo activity of B / T lymphocyte weakening factor immunogen polypeptide was detected by human small cell lung cancer tumor model.

[0019] 6-8w old C57BL / 6 nude mice were randomly divided into 4 groups, 50 males and 50 males, 10 mice in each group. (1) blank group; (2) low-dose polypeptide group; (3) medium-dose polypeptide group; (4) high-dose polypeptide group. The human 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, set three doses of polypeptide in the experimental group: 1.0, 2.0, and 4.0 mg / Kg, and inject 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 can improve the survival rate of tumor-bearing mice at doses of 1.0, 2.0, and 4.0 mg / Kg, a...

Embodiment 3

[0021] 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.

[0022] The experiment set up a blank control group, a model group (concanavalin A, purchased from sigma company), and a group of polypeptide dosages (5.0, 10.0, 20.0 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 5 μg / ...

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Abstract

The invention discloses a B and T lymphocyte attenuator immunogen polypeptide, belongs to the field of anti-cancer vaccines, and particularly relates to optimized polypeptide of B and T lymphocyte attenuator immunogen serving as an anti-cancer vaccine, and T cell immune responses are enhanced. The amino acid sequence is a brand-new sequence. The invention relates to application of the polypeptide in preparation of drugs for treating tumors, in particular to application in preparation of drugs which are used for tumor immune cells and used for enhancing the T cell immune responses and application in preparation of drugs for CAR-T cell immunotherapy. The B and T lymphocyte attenuator immunogen polypeptide has the advantages that the immunogen polypeptide has the brand-new amino acid sequence, can be used in tumor immune cell technologies, can promote T cell proliferation, can promote specific binding of T cells and tumor surface antigens, can inhibit tumor growth in tumor-burdened mouse bodies and is applied to cell therapies of CAR-T and the like by serving as target molecules of the 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 an immunogen of B / T lymphocyte weakening factor in an anticancer vaccine to enhance T cell immune response. 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] B / T cell weakening molecule (BTLA) is a recently discovered co-inhibitory molecule of the B / T lymphocyte w...

Claims

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Application Information

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IPC IPC(8): C07K14/705A61K39/00A61P35/00
CPCA61K39/0005C07K14/70521
Inventor 罗瑞雪
Owner SUZHOU PULUODA BIOLOGICAL SCI & TECH
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