Preparation and application of CpG DNA molecule anti-infection and immunity prepn

An immune preparation, anti-infection technology, applied in genetic material components, sugar derivatives, gene therapy, etc., can solve problems such as low efficiency

Inactive Publication Date: 2005-11-09
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since intramuscular injection mainly induces immune response through Th1 cell activation, the efficiency is low, and a large dose of DNA needs to be injected, while a larger amount of DNA is required through the digestive tract for oral administration

Method used

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  • Preparation and application of CpG DNA molecule anti-infection and immunity prepn
  • Preparation and application of CpG DNA molecule anti-infection and immunity prepn
  • Preparation and application of CpG DNA molecule anti-infection and immunity prepn

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0058] 1. Design and synthesis of CpG oligonucleotide sequence and its primers

[0059] CpG sequence design and synthesis:

[0060] CpG sequence (see sequence table on page 11 of the manual)

[0061] The sequence is 88 bases in length, contains 11 CpG motifs, and is characterized by containing 3 5’- AACGTT- 3’, 3 5’- GTCGTC- 3’, 1 5’- GACGTT- 3’, 1 5’- ATCGAT- 3’, 1 5’- GTCGTT- 3’, 1 5’- GGCGTT- 3’, 1 5’- GACGTC- 3'feature sequence.

[0062] CpG sequence primer design and synthesis:

[0063] 5’ end: CGAGATCTAACGTTGTC

[0064] 3’ end: CGAGATCTAACGTTAAC

[0065] 2. Preparation of CpG Oligonucleotides for Immunization

[0066] Using the synthesized CpG sequence as a template, it was amplified by PCR (polymerase chain reaction), and the amplified product was dissolved in sterile saline, and its concentration was measured with an ultraviolet spectrophotometer.

[0067] 3. CpG oligonucleotide stimulates the proliferation of animal blood immune cells in vitro

[0068] Under asepti...

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Abstract

An anti-infectious immunopotentiator for pig, ox, yak, etc is prepared through artificially synthesizing the CpG oligonucleotide sequce able to excite the reproductive activity of immune cell, preparing the chitosan nanoparticles from deacetyl chitosan, and using said chitosan nanoparticles for molecular packaging of CpG oligonucleotide. It can be used to immunize the experimental animal by intramuscular injection or oral applialtion.

Description

Technical field [0001] 1. Biotechnology 2. Molecular Immunology Background technique [0002] In 1984, Tokunaga et al. discovered that the DNA of Mycobacterium tuberculosis has anti-tumor properties in addition to being a kind of genetic material. This opened a new chapter in the study of bacterial DNA with immunostimulatory properties. Later, they thought through some studies This immunostimulatory property is caused by the hexanucleotide with self-complementary palindrome structure. By 1995, Krieg et al. found that bacterial DNA and some artificially synthesized ODN (oligodeoxynucleotides) have immunostimulatory effects, but this effect is not necessarily related to whether it has a palindrome structure, but is related to these effects. The unmethylated CpG structural unit (motif) in DNA is closely related and is defined as the CpG immunostimulatory sequence (CpG). [0003] CpG sequences are mainly found in the genomes of bacteria, viruses and invertebrates, but less in vertebr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00
Inventor 武梅高荣李惠付满良吴凯源杨毅章欢程驰刘狄广李化
Owner SICHUAN UNIV
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