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

Method for promoting immunological activity of gp96 protein and application thereof

A protein and immunomodulator technology, applied in the field of promoting the immune activity of gp96 protein, can solve the problems of ineffective stimulation of tumor cell immune response, loss of tumor antigens, changes, etc.

Inactive Publication Date: 2012-05-23
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
View PDF4 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Tumor cells often lose the expression of tumor antigens or change their antigen expression profile, so that tumor vaccines designed against known tumor antigens cannot effectively stimulate the immune response against tumor cells

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
  • Method for promoting immunological activity of gp96 protein and application thereof
  • Method for promoting immunological activity of gp96 protein and application thereof
  • Method for promoting immunological activity of gp96 protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Preparation of the extract of embodiment 1, N355 fragment, gp96 protein and gp96 protein

[0034] 1. Preparation of N355 Fragment

[0035] Using Escherichia coli to express the N355 fragment, the specific method is as follows:

[0036] 1. Design the following N355 primer pair, and the primers will be synthesized by Shanghai Yingjun Company:

[0037] Upstream primer: 5'-CGCGGATCCGACGATGAAGTTGATGTGGAT-3';

[0038] Downstream primer: 5'-CCGCTCGAGTTAAGTAAAGTGAATATAAGCCATG-3'.

[0039] 2. Extract the mRNA of human liver cancer cell HepG2 (purchased from ATCC, product number HB-8065), and synthesize cDNA by reverse transcription.

[0040] 3. Using the cDNA in step 2 as a template, perform PCR amplification with the N355 primer pair to obtain a PCR amplification product (coding sequence containing the N355 fragment).

[0041] 4. Digest the PCR amplification product with restriction endonucleases BarnHI and Xho I, and recover the digested product.

[0042] 5. Digest pGEX-6...

Embodiment 2

[0084] Embodiment 2, rgp96 protein and gp96 extract immune activity comparison

[0085] BALB / c mice were randomly divided into four groups, 10 mice in each group, and the following immunizations were carried out respectively (immunization volumes were equal):

[0086] The first group (HBc87-95): immunize mice with HBc87-95, 50 micrograms / time;

[0087] The second group (HBc87-95+gp96): the gp96 extract prepared in step 3 of Example 1 was used to immunize mice together with HBc87-95, the gp96 extract was 20 micrograms / time, and the HBc87-9550 micrograms / time;

[0088] The third group (HBc87-95+rgp96): immunize mice with the rgp96 protein prepared in step 2 of Example 1 and HBc87-95, rgp96 protein 20 micrograms / time, HBc87-9550 micrograms / time;

[0089] The fourth group (HBc87-95+IFA): mice were immunized together with incomplete Freund's adjuvant and HBc87-95, Freund's incomplete adjuvant 20 micrograms / time, HBc87-9550 micrograms / time.

[0090] Immunization was performed once...

Embodiment 3

[0091] Embodiment 3, the expression vector carrying gp96 gene and the immune activity comparison of gp96 extract

[0092] The coding sequence of the gp96 protein was inserted between the EcoR I and Xho I restriction sites of the expression vector pCDNA3.1 (purchased from Invitrogen, product number V790-20) to obtain a recombinant plasmid.

[0093] BALB / c mice were randomly divided into five groups, 10 mice in each group, and the following immunizations were carried out respectively (immunization volumes were equal):

[0094] The first group: immunize mice with HBc87-95, 50 micrograms / time;

[0095] The second group: immunize mice with recombinant plasmid and HBc87-95 together, recombinant plasmid 20 micrograms / time, HBc87-9550 micrograms / time;

[0096]The third group: immunize mice with recombinant plasmid and HBc87-95 together, recombinant plasmid 50 micrograms / time, HBc87-9550 micrograms / time;

[0097] The fourth group: immunize mice with recombinant plasmid and HBc87-95 t...

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 discloses a method for promoting immunological activity of gp96 protein and application thereof. The invention provides application of immunomodulators in the improvement of the immunological activities of gp96 protein-related maters; the immunomodulators are at least one of cyclophosphane, anti-CD25 antibody and an interfering RNA (Ribonucleic Acid) of Foxp3; the gp96 protein-related maters are the gp96 protein shown by a sequence 1 of a sequence table, N355 fragments or gp96-protein-contained extracts; and the interfering RNA of the Foxp3 is shown by a sequence 5 of the sequence table. The invention further provides various vaccines which comprise the immunomodulators and the shown gp96 protein-related maters. The invention has a great value to the treatment of various cancers, such as hepatitis B, melanoma, and the like.

Description

technical field [0001] The invention relates to a method for promoting the immune activity of gp96 protein and its application. Background technique [0002] Heat shock proteins (heat shock proteins, HSPs) have unique immunological functions, and its immunological mechanism includes two aspects: one is that HSPs participate in T cell antigen presentation; the other is that HSPs effectively stimulate innate immunity. [0003] Heat shock proteins exist in the cytoplasm and endoplasmic reticulum, and consist of multiple members, such as HSP60, HSP70, HSP90 and gp96. On the one hand, heat shock proteins play an important role as molecular chaperones in the process of protein folding and transport; on the other hand, they can bind to 5-25mer antigen polypeptides in cells, enter cells through CD91 molecules on the surface of antigen-presenting cells (APCs), and bind the antigens Epitopes are presented to MHC class I and II molecules, thereby initiating specific T cell immune resp...

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
IPC IPC(8): A61K39/39A61K39/29A61K39/00C07K1/18A61P1/16A61P31/20A61P35/00
Inventor 孟颂东李星辉刘振闫晓丽王赛锋李扬邱立鹏陈立钊胡坤
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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