Trichinella spiralis paramyosin h-2d restricted TH epitope p2, composition and application thereof

A composition and technology of Trichinella, applied in the field of immunobiology, can solve problems such as difficulty in correct diagnosis, difficulty in drug treatment, and inability to solve repeated infection of the worm

Active Publication Date: 2019-08-16
CAPITAL UNIVERSITY OF MEDICAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the complex clinical symptoms of trichinellosis, including diarrhea, fever, myalgia, edema, etc., it is difficult to diagnose correctly, which makes timely drug treatment difficult and drug treatment cannot solve the problem of repeated infection of the worm (Gottstein B, Pozio E, Nockler, K. Epidemiology, diagnosis, treatment, and control of trichinellosis. [J]. Clin Microbiol Rev, 2009, 22(1): 127-145.)

Method used

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  • Trichinella spiralis paramyosin h-2d restricted TH epitope p2, composition and application thereof
  • Trichinella spiralis paramyosin h-2d restricted TH epitope p2, composition and application thereof
  • Trichinella spiralis paramyosin h-2d restricted TH epitope p2, composition and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1: Preparation of rTs-Pmy protein

[0054] The Ts-Pmy plasmid bacterial liquid is transformed into Escherichia coli with a full-length plasmid containing the paramyosin gene (Ts-Pmy GenBank accession No.: EF429310), which is equivalent to the patent application with publication number CN100999737A (application number 200710000018.1) The Escherichia coli BL21 transformed with the paramyosin gene (Ts86cDNA) prepared in Example 2 can also refer to this patent application for its preparation method. Refer to this patent application to prepare recombinant Ts-Pmy protein (rTs-Pmy protein).

Embodiment 2

[0055] Embodiment 2: the synthesis of polypeptide sequence

[0056] 12 polypeptide sequences were synthesized according to the amino acid sequence of the recombinant Ts-Pmy protein (rTs-Pmy protein), as shown in Table 1 below. Wherein the position refers to the amino acid position on the rTs-Pmy protein.

[0057] Entrust Beijing Ovia Biotechnology Co., Ltd. to synthesize.

[0058] Table 1: Peptide sequences

[0059]

[0060]

Embodiment 3

[0061] Embodiment 3: In vitro test identification of polypeptide

[0062] 1. Experimental samples:

[0063] The recombinant Ts-Pmy protein (rTs-Pmy) prepared in Example 1.

[0064] Polypeptides P1-P12 synthesized in Example 2.

[0065] 2. Experimental animals and groups:

[0066] BALB / c female mice aged 6-8 weeks were randomly divided into two groups, the recombinant protein group and the PBS control group, with 6 mice in each group.

[0067] 3. Experimental method

[0068] (1) Immunization of animals: 50 μg of rTs-Pmy (dissolved in 50 μl PBS) or 50 μl of PBS plus an equal volume of adjuvant MONTANIDE per mouse TM ISA 50 V2 (Seppic Company), subcutaneously multi-point immunized mice, and boosted immunization with the same dose twice after 2 weeks and 4 weeks.

[0069] (2) Proliferation and isolation of spleen lymphocytes:

[0070] One week after the last immunization, the mice were killed and splenic lymphocytes were isolated (operated according to the instructions of...

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Abstract

The invention belongs to the field of immunobiology, and relates to H-2d restricted Th epitope P2 of trichina paramyosin, its composition and an application thereof. The invention concretely relates to Th2 epitope of trichina paramyosin, which contains an amino acid sequence shown in any one of SEQ ID NO: 2-5. The epitope of the present invention can stimulate / promote significant proliferation of splenic lymphocyte, can stimulate / promote cells increase of mice spleen cell CD4<+>T, can stimulate / promote differentiation of Th0 cell to Th2, can stimulate / promote increase of Th1-type cytokine IFN-gamma, IL-2 and Th2-type cytokine IL-4, IL-5, and can stimulate / promote differentiation of Th0 cell to Th2, after mice is immunized, a Th2 immunization reaction is generated through induction, and the epitope has potential for preparing vaccines such as multi-epitope vaccine.

Description

technical field [0001] The invention belongs to the field of immunobiology, and relates to a trichinella paramyosin Th2 epitope, such as trichinella spiralis paramyosin H-2d restricted Th epitope P2. The invention also relates to its composition and application. Background technique [0002] Trichinellosis is a zoonotic parasitic disease with global distribution. Human infection mainly comes from the highly pathogenic Trichinella spp. (Pozio E. World distribution of Trichinella spp. infections inanimals and humans. [J]. Vet Parasitol, 2007, 149(1-2): 3-21.) . Due to the complex clinical symptoms of trichinellosis, including diarrhea, fever, myalgia, edema, etc., it is difficult to diagnose correctly, which makes timely drug treatment difficult and drug treatment cannot solve the problem of repeated infection of the worm (Gottstein B, Pozio E, Nockler, K. Epidemiology, diagnosis, treatment, and control of trichinellosis. [J]. Clin Microbiol Rev, 2009, 22(1): 127-145.). Th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/435C12N15/12C07K19/00A61K39/00A61P33/10A01N47/44A01P5/00C07K16/18A61K39/395
Inventor 诸欣平顾园杨静黄京京
Owner CAPITAL UNIVERSITY OF MEDICAL SCIENCES
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