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Method for optimized preparation of inactivated vaccine and/or attenuated live vaccine

A technology of attenuated live vaccines and inactivated vaccines, applied in the field of preparation of whole virus vaccines, can solve the problems of low immunogenicity, lack of in-depth understanding of viruses, and failure to achieve protective effects

Inactive Publication Date: 2016-02-24
王晓佳
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the disadvantage of these two types of vaccines is that the immunogenicity is low and not comprehensive, and it needs to be used in combination with an adjuvant to produce a good immune effect, so the best protective effect is often not achieved
The above scheme has improved the titer of the vaccine virus to a certain extent, but it has not found and developed intracellular resources that are conducive to virus replication through an in-depth understanding of the host cell factors of virus replication

Method used

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  • Method for optimized preparation of inactivated vaccine and/or attenuated live vaccine
  • Method for optimized preparation of inactivated vaccine and/or attenuated live vaccine
  • Method for optimized preparation of inactivated vaccine and/or attenuated live vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 The half-infection of cells and the half-infection of chicken embryos of several viruses

[0032] 9-11-day-old SPF chicken embryos were taken to prepare chicken embryo fibroblasts (CEF), CDV, MDV and IBV were inoculated on CEF for 48 hours, harvested and the culture supernatant was repeatedly frozen and thawed 3 times, and then inoculated on CEF cells again Continue to culture, and so on to 3 generations. The virus solution was diluted 10 times and inoculated into 96-well plates or chicken embryos. CDV measurement of cell half-infectious dose (TCID) 50 ), MDV used cytopathic plaque formation to measure TCID 50 , IBV uses the half-infectious dose of chicken embryos EID 50 , to determine the virulence of three viruses. The results are shown in Tables 1-3.

[0033] Table 1 The influence of the half-infected amount of CDV cells (indirect immunofluorescence method)

[0034]

[0035] Calculated according to Reed and Muench formula: distance ratio=(lesion ra...

Embodiment 2

[0043] Example 2 Effect of knockout and overexpression of P60 on virus virulence (immunoblotting, lesion method and RT-PCR method)

[0044] 1. Preparation of cells

[0045] Passaging Hep-2 and Vero cells: the growth medium is DMEM medium containing 5% fetal bovine serum and 100U penicillin-streptomycin, and the maintenance medium is DMEM medium containing 2% fetal bovine serum and 100U penicillin-streptomycin.

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Abstract

The present invention provides a method for optimized preparation of an inactivated vaccine and / or an attenuated live vaccine, wherein in host cells, the cancer inhibition candidate gene 2 of glioma is subjected to over-expression to optimize the inside-host-cell replication environment for preparing the inactivated vaccine and / or attenuated live vaccine so as to improve the titers of 4 categories of viruses (including coronavirus, Paramyxoviridae, Orthomyxoviridae and herpes virus) by 1-3 lg unit. According to the present invention, the new strategy and the feasible basis are provided for the optimization of the whole virus vaccine virus titer, and important application prospects and innovative significance are provided in the biomedical field.

Description

technical field [0001] The invention relates to the field of preparation of whole virus vaccines, in particular to a method for optimizing the preparation of inactivated vaccines and / or live attenuated vaccines. Background technique [0002] Whole virus vaccines (including attenuated and inactivated vaccines) still occupy a considerable proportion of modern vaccine applications. Whole virus vaccines are also known as conventional vaccines, including inactivated vaccines and live attenuated vaccines. Live attenuated vaccines use artificial directed mutation methods, or select live microorganisms with highly attenuated or basically avirulent virulence from nature. made vaccines. After inoculation, the live attenuated vaccine has a certain ability to grow and reproduce in the body, and the immune effect is strong and lasting. Generally, it only needs to be vaccinated once, and the immunity will be maintained for life. In addition to stimulating the body to produce cellular imm...

Claims

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Application Information

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IPC IPC(8): C12N7/00C12N5/10C12R1/93
Inventor 王晓佳
Owner 王晓佳
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