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Novel asthma polypeptide vaccine and preparation method thereof

A technology for vaccines and asthma, applied in the field of biomedicine, can solve the problems such as the absence of IL-13 polypeptide-related vaccines, and achieve the effects of breaking immune tolerance, small molecular weight, and convenient synthesis

Inactive Publication Date: 2017-05-31
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, IL-13 monoclonal antibodies and soluble IL-13 receptors have entered clinical phase II and III, but there is no IL-13 peptide-related vaccine yet

Method used

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  • Novel asthma polypeptide vaccine and preparation method thereof
  • Novel asthma polypeptide vaccine and preparation method thereof
  • Novel asthma polypeptide vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0098] Example 1 Screening and Determination of Asthma IL-13 Short Peptide in the Present Invention

[0099] By consulting the library and detecting the binding affinity of IL-13 antigen short peptides to MHC, as well as the interaction between IL-13 antigen short peptides and IL-13 receptors, the best of the new asthma vaccines can be established. IL-13 antigen short peptide.

[0100] Specific method: From the Allele Frequency Net Database (AFND, http: / / www.allelefrequencies.net / ) database, the coverage of asthma-related screening ranges from 13.5% (DRB1*09:01) to 1% (DRB1*14:05) DR epitopes of human leukocytes, covering 95.9% of the Chinese population in total, obtained 20 epitopes. As a vaccine candidate antigen polypeptide, it needs to have high affinity with MHC, so 20 cases of Chinese HLA-DRB1 epitopes were predicted by six commonly used Immune Epitope Database (IEDB, http: / / www.iedb.org) method to filter. Among the 20 epitopes, the epitopes that can bind to MHC class...

Embodiment 3

[0116] Example 3 Animal immunization experiments and antibody titer evaluation of four polypeptides constructed on the basis of E3

[0117] After determining E3 as the optimal epitope with the function of treating asthma among the three candidate IL-13 antigen short peptides, the present invention considers coupling different helper T cell epitope polypeptides to enhance IL-13 antigen short peptide Immunogenicity, so as to obtain the best candidate polypeptide for the treatment of asthma, to be developed as a vaccine. Because when determining the IL-13 antigen short peptide epitope, the coupled helper T cell DiTOX epitope peptide (AYNFVESIINLFQVVHNSYN). However, it is not sure that this epitope has the best effect of increasing the immunogenicity of short peptides of IL-13 antigen, so we screened three other epitope peptides: PADRE; TT8; TT9; respectively coupled with E3 to obtain TT8- E3(V4), TT9-E3(V5) and PADRE((V6) are used together with DiTOX-E3(V3) to evaluate the thera...

Embodiment 4

[0132] Example 4 The results of the alveolar lavage fluid (BALF) cell count and related cytokine detection results of the animal immunization experiment of four polypeptides constructed on the basis of E3:

[0133] See embodiment 3 for grouping experimental treatment.

[0134] BALF count detection method is as follows: when the mice are killed, use pre-cooled saline for alveolar lavage, collect about 1mL of alveolar lavage fluid; Resuspended in saline and sent for GLP detection to count eosinophils. The supernatant was detected using the relevant kit detection method.

[0135] IFN-γ: The detection kit is R&D Systems Mouse IFN-gamma (Catalog Number: VAL607)

[0136] IL-4: The detection kit is R&D Systems Mouse IL-4 (Catalog Number: VAL603)

[0137] IL-13: The detection kit is eBioscience Mouse IL-13 (Catalog Number: 88-7137)

[0138] At the same time, the eyeballs were picked to collect blood, and the supernatant was collected by centrifugation and stored at -20°C for the d...

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Abstract

The invention belongs to the field of biological medicines, and concretely relates to a novel asthma polypeptide vaccine and a preparation method thereof. The invention aims to solve the technical problem so as to provide a new effective selection for asthma treatment in the field. The technical scheme provided by the invention for solving the technical problem is characterized by connecting DiTOX polypeptide or PADRE polypeptide on a nitrogen end of E3 polypeptide to obtain a new fusion polypeptide, wherein the polypeptide can be used as a main active constituent for preparing the asthma vaccine. The polypeptide vaccine provided by the invention can effectively induce specific humoral immunity, can break immune tolerance, and has a better application prospect when being used as the polypeptide vaccine for asthma treatment.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a novel asthma polypeptide vaccine and a preparation method thereof. Background technique [0002] According to the World Health Organization, there are currently about 300 million people worldwide suffering from asthma. Asthma is the most common non-communicable disease in children. Asthma is not just a public health problem in high-income countries; it occurs in all countries, regardless of their level of development. Most asthma-related deaths occur in low- and lower-middle-income countries. [0003] Bronchial asthma (Bronchial asthma, referred to as asthma) is a major non-communicable disease, characterized by dyspnea and recurrent wheezing, it is composed of a variety of cells (such as eosinophils, mast cells, T lymphocytes, middle Chronic inflammatory diseases of the airway involving granulocytes, airway epithelial cells, etc.) and cellular components. This chroni...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00A61K39/00A61P11/06
CPCA61K39/00C07K14/5437C07K14/705C07K2319/00
Inventor 杨莉何谷万洋魏于全
Owner SICHUAN UNIV
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