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Method capable of renforving gene gun inoculation DNA Vaccine inducel cell immune response

A DNA vaccine and cellular immunity technology, which can be applied in the field of enhancing the cellular immune response induced by DNA vaccine inoculation by gene gun, and can solve the problems of inhibition, antigenic weight reduction, influence and so on.

Inactive Publication Date: 2004-07-21
FUDAN UNIV +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the case where CpG-ODN is mixed with plasmid DNA and inoculated with a syringe, Weeratna et al. reported (Weeratna R et al. Antisense Nucleic Acid Drug Dev1998; 8:351-6) that ODN and plasmid DNA may compete with each other for cell entry receptors, Therefore, it may lead to the reduction of the amount of expressed antigen and the inhibition of CpG effect
[0009] In addition, the role of CpG-ODN in gene gun inoculation of DNA vaccines will also be affected by the ratio of CpG-ODN to plasmid DNA
Previous studies have used the ratio of CpG-ODN to plasmid DNA at 50 μg / 1 μg, but a large amount of CpG can strongly stimulate the secretion of cytokines and cause the expression of antigen genes in the plasmid to be inhibited (Tan Y et al. Hum Gene Ther1999; 10: 2153-61 )

Method used

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  • Method capable of renforving gene gun inoculation DNA Vaccine inducel cell immune response
  • Method capable of renforving gene gun inoculation DNA Vaccine inducel cell immune response
  • Method capable of renforving gene gun inoculation DNA Vaccine inducel cell immune response

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Embodiment 1

[0036] DNA was coated on gold powder with a diameter of 1.0 μm by spermidine-calcium chloride coating method to prepare gene gun bullets. Mix 10 μg plasmid DNA with 100 μg or 10 μg ODN, the weight ratio can be controlled at 5-10:1 or 1:1, or dissolve 10 μg plasmid DNA in 100 μl ddH alone 2 O, add to 100μl 0.1M spermidine containing 5mg gold particles and mix well, then add 200μl 2.5M CaCl 2 DNA was precipitated onto gold particles. After fully washed with ethanol, add 100 μl absolute ethanol and mix well, each 10 μl is a bullet. The gold particle-DNA ethanol solution is sucked into the special plastic tube of the gene gun, and the bullet is made after air drying. Among them, each bullet carries 0.5mg gold particles, 1μg DNA (8.34×10 -7 μmol), 10μg ODN (2.2×10 -3 μmol); or 0.5mg gold particles, 1μg DNA (8.34×10 -7 μmol), 1μg ODN (2.2×10 -4 μmol); or 0.5mg gold particles, 1μg DNA (8.34×10 -7 μmol).

[0037] The 6-week-old BALB / c female mice were immunized and randomly di...

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Abstract

A method for inducing the Th1-type immune response for cells after the gene gun is used to inoculate DNA vaccine features that when the plasmid DNA vaccine is inoculated by gene gun, the synthetic oligonucleotide CpG DNA (CpG ODN) as adjuvant is proportionally introduced. A process for preparing the plasmid DNA-CpG oligonucleotide coated gold particle is also disclosed. It can be used for treating tumor and durable virus infection.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a new method for enhancing the cellular immune response induced by gene gun inoculation with DNA vaccines, and its application. Background technique [0002] DNA vaccine, also known as gene vaccine, is essentially to clone exogenous protein-coding gene fragments on eukaryotic plasmid DNA expression vectors, and can express exogenous proteins in cells after direct inoculation and induce The body produces specific cellular and humoral immunity to prevent and treat diseases. Because it can overcome some of the defects of traditional vaccines, can induce the body to produce specific cellular and humoral immunity at the same time, and has the advantages of simple preparation and stable properties, it is considered to be a subunit vaccine following attenuated, inactivated vaccines and genetically engineered vaccines. The subsequent third-generation vaccine is a development direction of futu...

Claims

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Application Information

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IPC IPC(8): A61K48/00
Inventor 袁正宏周晓辉
Owner FUDAN UNIV
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