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Helicobacter pylori antigen HLA restrictive immunodominance epitope peptide and preparation method and application thereof

A Helicobacter pylori and dominant epitope technology, applied in the field of medical biology, can solve the problems of unavoidable epitopes, missed screening, and low accuracy

Active Publication Date: 2012-10-24
ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Currently, the reported Helicobacter pylori antigen CD4 + T cell epitopes are all mouse H-2 restricted Th epitopes. Due to the obvious differences between mouse and human MHC molecules, H-2 restricted epitopes may not be suitable for human vaccine design, so they need to be screened Identification of HLA-restricted epitopes
In addition, most of the currently reported epitopes of Helicobacter pylori are predicted by bioinformatics software, and the accuracy rate is not high. Although it can be verified by experiments, the phenomenon of epitope leakage cannot be avoided.

Method used

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  • Helicobacter pylori antigen HLA restrictive immunodominance epitope peptide and preparation method and application thereof
  • Helicobacter pylori antigen HLA restrictive immunodominance epitope peptide and preparation method and application thereof
  • Helicobacter pylori antigen HLA restrictive immunodominance epitope peptide and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 : Synthesis of 18 amino acid short peptides with overlapping steps and preparation of mixed peptide library

[0044] Helicobacter pylori flagellar urease protein B subunit (UreB), the sequence is conserved among various strains, with a full length of 569 amino acids, and the recombinant Helicobacter pylori urease B subunit is a 1-569 polypeptide derived from 11637 International Standard Strains. Therefore, the protein sequence of Helicobacter pylori flagella urease protein B subunit protein sequence (number P69996) derived from Helicobacter pylori 11637 was searched in the UniProt protein database, and a short peptide with 18 amino acid steps overlapping was synthesized from the first amino acid (provided by Shanghai Jier Biochemical Co., Ltd. The company assisted in the synthesis), a total of 93 (the last one is a 17 amino acid short peptide). The purity is greater than 70%. The synthetic peptide information is shown in Table 1. The synthesized peptides ...

Embodiment 2

[0054] Example 2 : Collection and Preservation of Peripheral Blood Mononuclear Cells from Patients Positive for Helicobacter pylori Infection

[0055] Collection and Preservation of Peripheral Blood Mononuclear Cells from Helicobacter Pylori Infection Positive Patients

[0056] The peripheral blood of patients with positive Helicobacter pylori infection was obtained from the Chongqing Blood Station of the Chinese People's Liberation Army, and was treated with C before blood collection. 13 For those who were positive for Helicobacter pylori infection in the urease breath test, the buffy coat cells were collected after blood collection, and then the peripheral blood mononuclear cells were separated with Ficoll-diatrizoate lymphocyte separation medium (Tianjin TBD Company), according to the actual instructions conduct. The isolated peripheral blood mononuclear cells were resuspended to a cell density of 1×10 7 / mL, add 1mL / tube into a cryopreservation tube, put it in a freeze...

Embodiment 3

[0057] Example 3 : Antigen-specific CD4 in peripheral blood of patients with Helicobacter pylori infection + Efficient expansion of T cells in vitro

[0058] 1. Method for expanding antigen-specific T cells in vitro

[0059] Resuscitate a tube of peripheral blood mononuclear cells from patients with positive Helicobacter pylori infection, and adjust the concentration of peripheral blood mononuclear cells in patients with positive Helicobacter pylori infection to 2.5×10 6 / ml, inoculate in 48-well cell culture plate (1ml / well), add appropriate amount of UreB antigen, mix well and incubate at 37°C, 5% CO 2 cultivated under conditions. On the fifth day, a low dose of recombinant human IL-2 (rhIL-2) (final concentration 25 U / ml) was added. On the 8th day, the medium began to turn yellow, and half of the medium was changed (the culture medium contained 25U / ml rhIL-2, and the cells were subcultured in time.

[0060] 2. ICS method to detect UreB-specific CD4 in peripheral blood...

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Abstract

The invention relates to Helicobacter pylori antigen HLA restrictive immunodominance epitope peptide as well as a preparation method and application thereof. The dominance epitope peptide has the amino acid sequences shown in SEQ ID NO:63, 74, 95 and 105. The invention also provides a preparation method of the epitope peptide, and further provides application of the epitope peptide to preparation for preventing or treating Helicobacter pylori infection.

Description

technical field [0001] The invention belongs to the technical field of medicine and biology, and relates to a Helicobacter pylori antigen HLA-restricted immunodominant epitope peptide and a preparation method and application thereof. Background technique [0002] In 1982, Australian scholars Warren and Marshall discovered a microaerophilic, Gram-negative, Helicobacter pylori (Helicobacter pylori, Hp) colonized in the human stomach and duodenum. Helicobacter pylori is the main pathogenic bacteria of the upper gastrointestinal tract, and is closely related to the occurrence of diseases such as gastritis, peptic ulcer, gastric mucosa-associated lymphoid tissue lymphoma (MALT) and gastric cancer. Studies in recent years have shown that Helicobacter pylori infection is also associated with extragastric diseases such as ischemic heart disease, atherosclerosis, and iron deficiency anemia. In 1994, the World Health Organization listed it as the first carcinogen in the population. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08A61K39/02A61P31/04C07K1/14
Inventor 吴超邹全明郭红杨武晨陈立李滨赵卓章金勇李海波毛旭虎郭刚童文德鲁东水
Owner ARMY MEDICAL UNIV
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