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SARS coronary virus resistant cell vaccine and its uses

A coronavirus and cell technology, used in antiviral agents, respiratory diseases, antibody medical components, etc., can solve the problems of poor vaccine antigenicity, short half-life, hidden safety hazards, etc., to promote killing effect, strong antigen processing and delivery. the effect of ability

Inactive Publication Date: 2007-07-04
THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the lack of correct post-translational processing and short half-life, the vaccines made of this protein often have poor antigenicity and cannot make the body produce a good protective immune response; DNA vaccines are simple to prepare and have a short cycle. With the release of the full genome sequence of the SARS virus, it is possible to prepare a DNA vaccine. The SARS vaccine announced by Shanghai to be released soon is a DNA vaccine. Although the DNA vaccine has many advantages, its safety is still a recognized hidden danger, especially for the The safety of the SARS virus with a high tendency to mutate is more worthy of consideration

Method used

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  • SARS coronary virus resistant cell vaccine and its uses
  • SARS coronary virus resistant cell vaccine and its uses
  • SARS coronary virus resistant cell vaccine and its uses

Examples

Experimental program
Comparison scheme
Effect test

example 2

[0159] Example 2: Mixed lymphocyte culture experiment detects that the cell vaccine against SARS coronavirus directly activates human T lymphocytes

[0160] Take 20ml of peripheral blood from healthy people, anticoagulate with heparin, place the blood in a 50ml plastic centrifuge tube under aseptic conditions, add an equal volume of 0.01MPBS to dilute, and add 20ml of lymphocyte separation solution to two 50ml plastic tubes, gently Gently add the above 20ml of diluted whole blood to the lymphocyte separation medium to keep the interface between the lymphocyte separation medium and the blood sample clear, centrifuge at 2500rpm for 30min, take out the centrifuge tube, and gently suck out the buffy coat layer with clear boundaries into another plastic tube , give 0.01MPBS, centrifuge at 1500rpm for 7min to wash the lymphocytes, wash repeatedly 4 times until the washing solution is clear, then add Hanks solution to suspend the cells, centrifuge at 1000rpm for 5min, wash the cells t...

example 3

[0164] Example 3: Detection of normal cell killing activity of mouse T lymphocytes expressing SARS gene fragments after immunization

[0165]Male BALB / C mice aged 6-8 weeks were randomly divided into experimental group and control group (ten mice in each group), and each mouse in the experimental group was intraperitoneally injected with 200ul (containing 1×10 6 ) anti-SARS coronavirus cell vaccine, and the control group mice were intraperitoneally injected with 200ul0.9% normal saline, once a week, 4 times in total. Collect mouse T lymphocytes immunized with anti-SARS coronavirus cell vaccine as effector cells, wash 2 times with 1640 culture medium, count, adjust cell concentration to 1×10 6 / ml. Normal human primary cultured vascular endothelial cells infected with SARS gene fragment recombinant adenovirus were used as target cells, and the cell concentration was adjusted to 1×10 5 / ml. Make 3 duplicate wells of effector cells, 100ul per well, add to 96-well U-shaped plat...

example 4

[0168] Example 4: Detection of normal human primary cultured vascular endothelial cell antibodies against SARS gene fragment recombinant adenovirus infection in mouse serum by flow cytometry

[0169] On the 5th day after the third immunization, the mice in the immunized group and the control group were blood collected through the orbit, and the serum of the mice was collected, and 1×10 6 Normal human primary cultured vascular endothelial cells infected by a recombinant adenovirus of SARS gene fragments, normal human primary cultured vascular endothelial cells, and normal human primary cultured vascular endothelial cells infected by empty adenovirus were transferred to flow cytometry sample tubes and washed with PBS. 100ul of sera from the mouse treatment group and the control group were added to each tube with a dilution ratio of 1:500. PBS solution was used instead of mouse serum as primary antibody blank control. Place in a refrigerator at 4°C in the dark for 1 hour, wash w...

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Abstract

The invention discloses an anti-SARS coronavirus cell vaccine and its application, said cell vaccine (1) containing all or part gene fragment of SARS coronavirus S protein all length gene order and 5 specific eukaryotic expression vector of cDNA fragment; (2) expressing the heterogenetic antigen presenting cells of said SARS coronavirus gene corresponding protei; (3) inactivating the heterogenetic antigen presenting cells expressing SARS coronavirus related protein. Use said cell vaccine for immuning host containing humans and rodents such as mice in order to making them generate protective humoral immunity and cellular immunity and resisting the SARS coronavirus infection which causes atypical pneumonia contagion. The security and stability of the invention are fine, production is convenient and price is low, it can induce the body generating the effective humoral immunity and cellular immunity of SARS coronavirus causing atypical pneumonia at the same time; provide effective and cheap vaccines for treating and preventing SARS.

Description

technical field [0001] The invention relates to the field of vaccines for preventing and treating atypical pneumonia caused by SARS coronavirus, in particular to a cell vaccine expressing SARS coronavirus S and five specific proteins. Also relate to the purposes of this vaccine in anti-SARS coronavirus. Background technique [0002] Severe Acute Respiratory Syndrome (SARS) is spreading in China as an emerging severe infectious disease. Effective prevention and treatment of SARS is the most important task of the country at present, but no very effective treatment drug has been found so far. Therefore, effective prevention and treatment of SARS is imminent. [0003] In terms of prevention and treatment, vaccine preparation is one of the most promising methods, which includes the following technologies: attenuated or inactivated virions, genetically engineered recombinant vaccines, polypeptide vaccines, and DNA vaccines. In addition to inducing the body to produce specific an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/215A61K45/00A61K48/00A61P31/14A61P11/00
Inventor 贺伟峰吴军
Owner THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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