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Method for evaluating immune effect of dengue vaccine

A technology for dengue vaccine and immune effect, applied in the field of immunology, can solve the problems of high cost, easy death, and inability to accurately simulate the immune effect of vaccines.

Active Publication Date: 2019-03-19
ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When using these two models to evaluate the immune effect of dengue vaccine, there are certain shortcomings. The former cannot accurately simulate the immune effect of the vac

Method used

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  • Method for evaluating immune effect of dengue vaccine
  • Method for evaluating immune effect of dengue vaccine
  • Method for evaluating immune effect of dengue vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] A kind of immune effect evaluation method of dengue vaccine, it comprises the following steps:

[0043] 1) At first utilize monolayer vero cell to cultivate dengue virus, the virus liquid that obtains after poisoning is measured titer by plaque test, the result is (6.25×10 5 ); then the dengue virus liquid was divided into 200 μl / tube, 20 μl / tube, 4 μl / tube and 2 μl / tube, 50 tubes each, and stored at -80°C for later use;

[0044] 2) Prepare a number of pregnant Balb / c mice, and use the virus in step 1) to challenge the newborn blank offspring intracranially; in order to measure the virus titer when 50% of the suckling mice die, it is necessary to test the blank newborn milk Rat intracranial challenge dengue virus of different titer groups, that is, culture stock solution group, 10-fold dilution group, 50-fold dilution group and 100-fold dilution group, of which 10-fold dilution group, 50-fold dilution group and 100-fold dilution group Add 180 μl, 196 μl and 198 μl of s...

Embodiment 2

[0058] A kind of immune effect evaluation method of dengue vaccine, it comprises the following steps:

[0059] 1) Firstly, a monolayer of vero cells was used to culture dengue virus, and the virus liquid obtained after harvesting the virus was tested for titer by plaque test (6.25×10 5 ), then the dengue virus solution was divided into 200 μl / tube, 20 μl / tube, 4 μl / tube and 2 μl / tube, 50 tubes each, and frozen at -80°C for later use;

[0060] 2) Prepare several pregnant Balb / c mice, and use the virus in 1) to challenge the newborn blank offspring intracranially. Intracranial challenge dengue virus of different titer groups, that is, culture stock solution group, 10-fold dilution group and 100-fold dilution group, in which 10-fold dilution group and 100-fold dilution group were added with 180 μl, 196 μl and 198 μl of no Bacteria PBS solution, every group intracranial injection dosage is 20 μ l each, the virus titer group that causes 50% suckling mice to die repeats 5 times, an...

Embodiment 3

[0073] A kind of immune effect evaluation method of dengue vaccine, it comprises the following steps:

[0074] 1) Firstly, a monolayer of vero cells is utilized to cultivate dengue virus, and the virus liquid obtained after receiving the virus is measured by a plaque test for titer (6.25×10 5 ), then the dengue virus solution was divided into 200 μl / tube, 20 μl / tube, 4 μl / tube and 2 μl / tube, 50 tubes each, and frozen at -80°C for future use.

[0075] 2) Prepare several pregnant Balb / c mice, and use the virus in 1) to challenge the newborn blank progeny mice intracranially. In order to measure the virus titer when 50% of suckling mice die, it is necessary to challenge dengue virus of different titer groups intracranially to blank neonatal suckling mice, i.e. culture stock solution group, 10-fold dilution group and 100-fold dilution group, wherein The 10-fold dilution group and the 100-fold dilution group were added with 180 μl and 198 μl sterile PBS solution before use, respec...

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Abstract

The invention discloses a method for evaluating the immune effect of a dengue vaccine. The method comprises the steps of carrying out toxicity attack on a newly born offspring suckling mouse of an immunized adult Balb/c mouse in an encephalic injection manner by virtue of the dengue virus, and observing and recording the death condition of the suckling mouse within 21 days after the toxicity attack so as to evaluate the immune effect of the vaccine. Compared with an existing method for evaluating the immune effect of the dengue vaccine by virtue of a wild suckling mouse model or a mouse modelwith immunologic function defects, the method has the advantages that the disadvantages of the two models are avoided, and the constructed method can be used for accurately evaluating the immune effect of the vaccine in an actual user object and is relatively economic, practical, simple and feasible.

Description

technical field [0001] The invention relates to the field of immunology, in particular to a method for evaluating the immune effect of a dengue vaccine. Background technique [0002] Dengue virus belongs to a subgroup of serotypes in the genus Flavivirus of the family Flaviviridae. It is divided into four serotypes 1, 2, 3, and 4 according to antigenicity. Dengue fever, dengue hemorrhagic fever, and dengue shock can occur after infection syndrome. These diseases are widely prevalent in tropical and subtropical regions, and are human infectious diseases with wide distribution, frequent incidence and great harm. Currently, about 3.9 billion people in 128 countries and regions are threatened by dengue virus. [0003] Many studies have been carried out on dengue virus, but the establishment of animal models of dengue virus is more challenging because humans are currently the only hosts infected with dengue virus and producing symptoms in nature. Animal models are an effective...

Claims

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Application Information

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IPC IPC(8): A61K49/00
CPCA61K49/0008
Inventor 李晋涛邱民月叶楠郭红霞杨茂成
Owner ARMY MEDICAL UNIV
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