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DNA vaccine carrier

A technology of DNA vaccine and carrier, applied in the field of DNA vaccine carrier construction, to achieve the effect of improving immune adjuvant effect and high copy number

Inactive Publication Date: 2004-09-01
SHANGHAI FUDAN YUEDA BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there is no vector for DNA vaccines identical to the present invention

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Example 1: The level of antibodies and spleen cells secreted cytokines in the body after vaccination with pYQF series vectors.

[0014] BALB / c mice were immunized with pYQF series vectors pYQF / HBs, pYQF-CpG / HBs, etc., which have been inserted into the hepatitis B virus surface antigen encoding gene. There were 12 mice in each immunization group. Using bilateral hind leg tibial anterior muscle injection method, each mouse was injected with 50μg / 100μl DNA physiological saline solution. 6 mice were immunized with pCI-neo / HBs and HBsAg protein (1μg / mouse), and an empty pYQF control and a negative mouse control (2 mice each) were set. The same dose was boosted once in the 4th week after the initial immunization. Blood was collected regularly at 2, 4, 6, and 8 weeks after the initial immunization, and the serum was separated and stored at -20°C. ELISA is also tested. The ELISA test results showed (Figures 7, 8) that pYQF / HBs, pYQF-1CpG / HBs and pYQF-2CpG / HBs can induce mice to pro...

Embodiment 2

[0017] Example 2: The level of antibodies induced by CpG ODN+HBsAg immunized mice.

[0018] HBsAg recombinantly expressed by yeast was mixed with synthetic CpG oligonucleotide (ODN) and Alum to immunize BALB / c mice, and HbsAg alone was used as a control. After 4 weeks of immunization, blood was collected, and the serum was diluted 1:50, and anti-HBs antibodies were determined by ELISA. The results show (Table 6), Oligo-1 (5′-AACGTT-3′), Oligo-4 (5′-AACGTC-3′), Oligo-5 (5′-AGCGCT-3′) and Oligo-7 ( 5'-GGCGTC-3') can increase the adjuvant effect of HBsAg inducing mice to produce specific antibodies, and has an immune adjuvant effect.

Embodiment 3

[0019] Example 3: The level of antibodies induced after pYQF-hCpG / HBs immunization.

[0020] BALB / c mice were immunized with plasmids pYQF-hCpG / HBs and pYQF / HBs, which were inserted into the gene encoding the hepatitis B virus surface antigen. 6 mice in each immunized group, each mouse was injected intramuscularly with 50 μg DNA. An empty pYQF control and a negative mouse control (2 mice each) were set up. The same dose was boosted once in the 4th week after the initial immunization. After the immune serum was diluted 1:2000 at the 8th week, the anti-HBs IgG antibody was determined by ELISA. The results show that( Figure 9) The A450 corresponding to the pYQF-hCpG / HBs immunization group was 1.12±0.47 (mean±SD, n=6), and the corresponding A450 value of the pYQF / HBs immunization group was 0.63±0.54 (mean±SD, n=6). The experimental results show that the CpG element 5'-AGCGCT-3' inserted into the pYQF / HBs vector has a certain stimulating effect on its ability to induce specific immune...

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Abstract

A novel DNA vaccine carrier for human body is composed of CMV promotor / enhancer, BGH transcription termination signal, multi-clone site, kanamycin resistance gene, immunostimulant of human immune system and exogenous protein coding gene, and can be used for preventing and treating diseases. The process for configuring the said DNA vaccine carrier and the oligonucleotide for effectively stimulating human immune system and screening then is also disclosed.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and relates to the construction of a DNA vaccine vector that is efficient, safe, and suitable for use in humans, and the practical use of this series of vectors. Background technique [0002] DNA vaccine (DNA vaccine) is also called gene vaccine. Its essence is to clone the foreign protein-encoding gene fragment on the eukaryotic plasmid DNA expression vector, and directly inoculate the body to express the foreign protein in the cell and induce the body Produce specific cellular and humoral immune responses to achieve the purpose of preventing and treating diseases. Because it can overcome some of the shortcomings of traditional vaccines, can simultaneously induce the body to produce specific cellular and humoral immune responses, and has the advantages of simple preparation and stable properties, it is considered as a secondary attenuated, inactivated vaccine and genetic engineering subunit vaccine The s...

Claims

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Application Information

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IPC IPC(8): A61K48/00C12N15/31C12N15/51C12N15/63
Inventor 袁正宏邱海军闻玉梅
Owner SHANGHAI FUDAN YUEDA BIOTECH
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