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Application of recombinant trichina thioredoxin peroxidase 2 in preparation of anti-trichina infection vaccine

A technology of thioredoxin and peroxidase, which can be used in anti-infective drugs, oxidoreductase, peptide/protein components, etc., and can solve the problem of no biological products.

Active Publication Date: 2020-03-24
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At this stage, many scholars are conducting research on immune prevention of Trichinella spiralis, but so far there is no applicable biological product

Method used

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  • Application of recombinant trichina thioredoxin peroxidase 2 in preparation of anti-trichina infection vaccine
  • Application of recombinant trichina thioredoxin peroxidase 2 in preparation of anti-trichina infection vaccine
  • Application of recombinant trichina thioredoxin peroxidase 2 in preparation of anti-trichina infection vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] rTsTPX2 induces macrophage differentiation in vitro

[0037] Material:

[0038] Raw 264.7 is a commercially available cell, purchased from the Cell Bank of the Chinese Academy of Sciences, catalog number: TCM13.

[0039] Balb / C mouse peritoneal macrophages: macrophages obtained from the peritoneal cavity of mice. Balb / C mice were purchased from the animal farm of Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences. The method of obtaining peritoneal macrophages: injected with 2 mL of PBS into the abdominal cavity of the mouse, and gently pat the abdominal cavity of the mouse. The PBS containing the cells was withdrawn with a needle, put into a 15 mL centrifuge tube, added 2 mL of erythrocyte lysate (Macgene, product number: CC051), and incubated at 4°C for 2 h. Then the lysate was centrifuged at 4°C and 1000rpm for 7min, the supernatant was discarded, and repeated twice. Resuspend the pellet with culture medium, according to 10 6 / well in...

Embodiment 2

[0047] rTsTPX2-induced activated macrophages activate CD4 + T cell

[0048] Take the spleen of healthy Balb / C mice and use CD4 + T Magnetic Bead Isolation Kit to separate CD4+ T cells and adjust CD4 + T cells to 5 x 10 6 / mL, the mouse peritoneal macrophage concentration after the stimulation of the experimental group in Example 1 was adjusted to 1×10 6 / mL. The two kinds were co-cultured for 72 hours in RPMI-1640 medium containing 5% (volume concentration) newborn bovine serum, 2mM L-glutamine, 100U / mL penicillin and 100μg / mL streptomycin.

[0049] Control group comprises positive control group, negative control group and blank control group; Positive control group is the macrophage stimulated with IL-4 in embodiment 1 and CD4 + T cell co-culture; Negative control is the macrophage stimulated with IFN-γ in Example 1 and CD4 + T cell co-cultivation; Blank control is the macrophage stimulated with PBS in embodiment 1 and CD4 + T cell co-culture.

[0050] MTs method to d...

Embodiment 3

[0052] Immunization of Balb / C mice with rTsTPX2 protein

[0053] Balb / C mice were subcutaneously immunized with 100 μL of rTsTPX2 protein (diluted in PBS) at a concentration of 0.5 mg / mL, and the second immunization was performed one week later (the same method and dose as the first immunization). One week after each immunization, the blood was collected for serum, and the whole blood of the mice was collected one week after the second immunization, and the peripheral blood lymphocytes were separated to detect the content of CD4+T and CD8+T lymphocytes.

[0054] The result is as image 3 As shown, where A-C is the ELISA detection of the change of IL-4 in rTsTPX2-immunized mice; D-F is the ELISA detection of the change of IFN-γ in rTsTPX2-immunized mice; G is the gating strategy to analyze the CD4 in peripheral blood lymphocytes + T cells and CD8 + T cells, where (I) label the appropriate T cells, (II) based on (I) CD3 + T cell gating, (III) based on (II) on CD8 + T cell ga...

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Abstract

The invention provides an application of recombinant trichina thioredoxin peroxidase 2 in preparation of an anti-trichina infection vaccine, which belongs to the technical field of immunoregulation, and the recombinant trichina thioredoxin peroxidase 2 can induce a mouse body to generate Th2 type immune response, induce proliferation of CD4<+> T cells of a mouse and convert macrophages into M2 type cells by the recombinant trichina thioredoxin peroxidase 2. The immune response reactions play a main role in the trichinella spiralis infection resisting process; therefore, the recombinant trichina thioredoxin peroxidase 2 as an antigen molecule can be used for preparing an anti-trichina infection vaccine.

Description

technical field [0001] The invention belongs to the technical field of immune regulation, and in particular relates to the application of recombinant Trichinella spiralis thioredoxin peroxidase 2 in preparing vaccines against Trichinella spiralis infection. Background technique [0002] Trichinella spiralis is a parasitic worm belonging to the Phylum Nemathelminthes class Nematoda, also known as Trichinella spiralis. Trichinosis can cause a serious disease of humans and other mammals (pigs, dogs, cats, bears, foxes, mice) called trichinosis. Trichinella spiralis mates in the small intestine of the host, and then the female burrows into the intestinal wall to produce larvae, which are carried to all parts of the body by lymph / blood. In muscle tissue, Trichinella spiralis larvae induce muscle cells to undergo a series of physiological and biochemical characteristics and structural reconstruction, making them lose the characteristics of muscle cells and form cysts. After the ...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61K39/39A61K38/44A61P37/04A61P33/10
CPCA61K38/443A61K39/0003A61K39/39A61K2039/55566A61P33/10A61P37/04C12Y111/01015
Inventor 张念章李文卉金启旺秦洪涛付宝权
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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