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CD8+T cell dominant epitopes based on toxoplasmagondii bradyzoite antigens

A dominant epitope and Toxoplasma gondii technology, applied in the direction of anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, protozoan antigen components, anti-animal/human immunoglobulin, etc., can solve the undiscovered prevention and control Drugs and other issues

Active Publication Date: 2013-09-04
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For the treatment of toxoplasmosis, no ideal control drug has been found so far

Method used

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  • CD8+T cell dominant epitopes based on toxoplasmagondii bradyzoite antigens
  • CD8+T cell dominant epitopes based on toxoplasmagondii bradyzoite antigens
  • CD8+T cell dominant epitopes based on toxoplasmagondii bradyzoite antigens

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Example 1 Screening of dominant epitopes, immunization of BALB / c mice

[0013] The SAG2C, SAG2D and SAG2X protein sequences were retrieved from the Toxoplasma gondii database (ToxoDB: http: / / toxodb.org / ). Antigenic epitopes were screened using the (ANN, SMM) algorithm in the Immunological Epitope Database (IEDB) (http: / / www.iedb.org / ). Screened out 6 H-2 (Ld, Kd, ​​Dd) high-affinity CD8 + The specific sequence of the T cell epitope peptide (Table 1) is shown in 1-6 in the sequence listing, and it is artificially synthesized. Dissolve the peptides in PBS and mix them to immunize BALB / c mice. Inject 100ul of the peptide mixture at the base of the tail of the mice, in which the content of each peptide is 100ug. Immunization 2 times, 3 weeks apart.

Embodiment 2

[0014] Example 2 Cell Proliferation Experiment of Immunized Mice

[0015] Four weeks after immunization, the spleen of the immunized mice was taken under aseptic conditions, the spleen cell suspension was prepared, and the epitope peptide was added to stimulate, at 37°C, 5% CO 2 After culturing under the condition for 48 hours, 10 μl of WST-8 dye was added to every 100 μl of cell culture medium, and the lymphocyte proliferation experiment was carried out by the CCK-8 method, and the absorbance value of each well was measured at 450 nm. (results see figure 1 ).

[0016] The results showed that the two epitope peptides: VPNSSLVEN and SQFLSLSLL showed high absorbance values, which proved that they had a strong ability to stimulate the proliferation of splenocytes, and could be used as superior candidate epitopes for the design of epitope vaccines.

Embodiment 3

[0017] Example 3 Determination of the production of cytokines induced by the dominant epitope vaccine in mice

[0018] After dissolving the polypeptide VPNSSLVEN and SQFLSLSLL in PBS to 10ug / ul, inject the immunized BALB / c mice into the tail of the mice, divide them into control group, single peptide immunization group and mixed immunization group, and the single peptide immunization group was injected with single peptide VPNSSLVEN100ug respectively Or SQFLSLSLL100ug, the mixed immunization group was injected with two kinds of single peptides at the same time, and the content of each peptide was 100ug. Immunization 2 times, 3 weeks apart. The preparation of the splenocyte suspension of immunized mice was as described in Example 2. After the splenocytes from mice were stimulated, the culture supernatants were collected at different times to measure the contents of IL-2 and IL-10 in cells. The results are shown in Table 2. A large amount of IL-2 was detected in the splenocyte ...

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Abstract

The invention discloses two CD8+T cell dominant epitopes of VPNSSLVEN and SQFLSLSLL based on toxoplasmagondii bradyzoite specific surface antigens, wherein six CD8+T cell epitopes with H-2 restriction and high affinity are screened according to the protein sequences of the bradyzoite specific surface antigens of SAG2C, -2C and -2X; BALB / c mice are actively immunized by the screened epitope polypeptides; determinations for a lymphopoiesis level and a cell factor content, and an immune protection evaluation for epitope vaccines are performed after the immunization. The result indicates that powerful cell immunization can be generated by inducing the mice via the epitope vaccinesm, wherein the two antigen peptides of VPNSSLVEN and SQFLSLSLL are provided to be great in the capacity of stimulating T cell proliferation, as well as capable of inducing the mice to secrete protective cell factors and immune protection, being used as the epitope vaccines effectively resisting toxoplasmagondii infection, and reducing an encystations rate in the brain tissue of a host.

Description

technical field [0001] The present invention relates to CD8 based on toxoplasma gondii antigen + T cell dominant epitope. Background technique [0002] Toxoplasmagondii is a worldwide distributed parasitic protozoan, which widely parasitizes in the nucleated cells of human and animals, and can cause great clinical damage to human health and huge economic losses in animal husbandry. For the treatment of toxoplasmosis, no ideal control drug has been found so far. The epitope vaccine has become a research hotspot of Toxoplasma gondii vaccine in recent years because of its easy synthesis, simple development and safety. Antigenic epitopes are antigenic determinants on the surface of protein antigens, which are special chemical groups that determine antigen specificity. Antigens bind to corresponding antigen receptors on the surface of lymphocytes through antigenic epitopes, thereby activating lymphocytes and causing immune responses; antigens also use epitopes to specifically ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C07K16/28A61K39/002A61P33/02
Inventor 丛华袁泉张敏丛海滋赵玲潇
Owner SHANDONG UNIV
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