Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

A mucosal immune adjuvant for inducing th1-type immune response and its application

A mucosal immune adjuvant and immune response technology, which is applied in the direction of medical preparations containing active ingredients, the use of carriers to introduce foreign genetic materials, hybrid peptides, etc., can solve the problem of poor immune protection, weak antigen immunogenicity, and immune Respond to problems such as short duration, achieve good social and economic benefits, broad market prospects, and good immune effects

Inactive Publication Date: 2014-10-15
PLA NAVY GENERAL HOSIPTAL
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In view of the current adjuvant research, the immunogenicity of the antigen is generally weak, the duration of the immune response induced by the mucosal immune vaccine is short, and the immune protection effect is not good. The present invention provides a method to improve the intensity of the immune response and change the type of immune response A new type of mucosal immune adjuvant that induces IgG2a antibodies that depend on Th1-type immune responses (Th1-type immune responses play an important role in anti-infection and anti-tumor responses)

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A mucosal immune adjuvant for inducing th1-type immune response and its application
  • A mucosal immune adjuvant for inducing th1-type immune response and its application
  • A mucosal immune adjuvant for inducing th1-type immune response and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Example 1 Determination of the specific binding active site between RTB and cells

[0073] 1. Using molecular simulation to construct the spatial conformation of the 3E1 light and heavy chain antibody

[0074] The amino acid sequence of the heavy chain of the antibody 3E1 with independent intellectual property rights is as follows: (see sequence 1 in the sequence listing)

[0075] S G A E L M K P G A S V K I S C K A T G Y T F S S Y W I E W I K Q R P G H G L E W I G D I L P G SG S T N Y N E K F K G K A T F T A D T S S N T A Y M Q L S S L T S E D S A V Y Y C S R S F Y Y N Y D GA Y F A Y W G Q G T L V T V S A A K T T P P S V Y P L A P G S A A Q T N S M V T L G C L V K G Y F P EP V T V T W N S G S L S S G V H T F P A V L Q S D L Y T ​​L S S S V T V P S S T W P S E T V T C N V A H PA S S T K V D K K I V P R D C

[0076] A template protein with high homology (PDB library number: 2V7H) was selected for sequence alignment, and the result ( figure 1 .). The antibody heavy chai...

Embodiment 2

[0090] Example 2. Construction, expression, purification and identification of GFP-nRTB fusion protein prokaryotic expression vector

[0091] 1. Materials

[0092] PCR kits, plasmid extraction kits, gel recovery kits, Lysozyme, DNA and protein markers are all products of Shanghai Sangong; PET-30a(+) prokaryotic expression vector, restriction enzymes Nco I, EcoRV, T4 DNA ligase and pGEM-T Easy were purchased from Promega. PCR primer synthesis and DNA sequencing were completed by Shanghai Boya Biotechnology Co., Ltd.

[0093] 2. Method results

[0094] 1. Construction and design of prokaryotic expression vector ( Figure 7 . Figure 8 .)

[0095] Use PET-30a (+) prokaryotic expression vector. After analysis by BIOSUN software, the enzyme cutting sites Nco I and EcoRV were selected. The protein is expressed from ATG, and the C-terminus of the recombinant protein has a stop codon.

[0096] 2. Design PCR primers according to the target gene sequence.

[0097] Codon optimiz...

Embodiment 3

[0117] Example 3. Preliminary Evaluation of GFP-nRTB Fusion Protein Mucosal Immune Effect

[0118] 1. Materials

[0119] IgG, IgG1, IgG2a, IgG2b, IgG3 monoclonal antibodies are all products of Sigma. HRP-labeled goat anti-mouse antibody (GAM-HRP) was produced by Boaosen Biotechnology Co., Ltd.

[0120] 2. Method results

[0121] 1. Preliminary evaluation of GFP-nRTB fusion protein mucosal immune effect

[0122]The purified four fusion proteins and 50 μg of GFP were orally administered sublingually to the mice with 0, 14, and 28 immunization procedures, and blood was collected on the 7th day after the third immunization, and the GFP IgG and immunoglobulin subunits in the serum were detected. For IgG1, IgG2a, IgG2b, and IgG3 levels, the serum used was diluted 1:100 with pH7.4 phosphate buffer and detected by indirect ELISA. The results showed that the levels of GFP IgG, IgG2b, and IgG3 in the serum after P1-GFP immunization were not significantly different from those of othe...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to a mucosal immunoadjuvant inducing Th1 immune response. A bioinformatics technology is applied in the invention, the spatial conformation of a ricin B-chain monoclonal anti-3E1 and Ricin-B compound which has obtained a Chinese invention patent is determined through the action of an antibody-antigen compound, and a molecular stimulation technology is utilized to reasonably determine four Ricin B identifying active regions of 3E1 and derive four nucleotide sequences. Four GFP-nRTB fusion proteins are obtained through a molecular cloning technology. Results of experiments that the four proteins are used to immunize BALB / C mice through a sublingual way three times show that a P1-GFP protein in the four proteins excites Th1 immune response, and a high-level IgG2a antibody is generated. The capability and the possible molecular mechanism of the targeting GFP mucosal immune response of the P1-GFP protein are discussed. The mucosal immunoadjuvant inducing Th1 immune response, which is screened through the above scheme, can be used for preparing vaccines, immunomodifiers or treatment medicines.

Description

technical field [0001] The invention relates to a mucosal immune adjuvant capable of directly targeting antigens to antigen-presenting cells, and its application in the preparation of vaccines, immunomodulators or therapeutic drugs. Background technique [0002] The mucous membrane is an important physiological barrier to prevent the invasion of pathogenic microorganisms in the natural immune system of the human body. More than 95% of the infections in the body occur in the mucous membranes or invade the body from the mucous membranes. The resulting mucosal damage and mucosal immune dysfunction often become an important cause of autoimmune diseases or opportunistic infections. Plays a pivotal role in human health [1] . [0003] Mucosal immunity can induce local mucosa to produce protective antibodies such as secretory IgA (sIgA), IgM and IgG, and induce other parts to produce sIgA and systemic humoral immune response; at the same time, mucosal immunity can also induce muco...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/06C07K19/00C12N15/11C12N15/63A61K39/39
Inventor 郭建巍秦力维冯健男
Owner PLA NAVY GENERAL HOSIPTAL
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com