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A method for optimizing the preparation of inactivated vaccines and/or live attenuated vaccines

A technology of attenuated live vaccines and inactivated vaccines, which is applied in the field of preparation of whole virus vaccines, and can solve problems such as lack of in-depth understanding of viruses, incompleteness, and low immunogenicity

Inactive Publication Date: 2019-02-19
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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  • A method for optimizing the preparation of inactivated vaccines and/or live attenuated vaccines
  • A method for optimizing the preparation of inactivated vaccines and/or live attenuated vaccines
  • A method for optimizing the preparation of inactivated vaccines and/or live attenuated vaccines

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Cell half infection dose and chick embryo half infection dose of several viruses

[0032] Take 9-11 day-old SPF chicken embryos to prepare chicken embryo fibroblasts (CEF), inoculate CDV, MDV and IBV on CEF and culture for 48 hours, harvest cells and culture supernatant repeatedly freeze-thaw 3 times, and re-inoculate on CEF cells Continue to cultivate, so pass down to 3 generations. Dilute the virus solution 10 times, and inoculate cells in 96-well plates or chicken embryos. CDV measured cell half infection dose (TCID 50 ), MDV uses the formation of cytopathic acne spots to measure TCID 50 , EID 50 , to determine the virulence of the 3 viruses. The results are shown in Table 1-Table 3.

[0033] Table 1 The influence of CDV cell half infection amount (indirect immunofluorescence method)

[0034]

[0035] Calculated according to the formula of Reed and Muench: distance ratio=(lesion rate higher than 50%-50) / (lesion rate higher than 50%-lesion rate lowe...

Embodiment 2

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

[0044] 1. Cell preparation

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

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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 vaccines and inactivated vaccines) still occupy a considerable proportion in modern vaccine applications. Whole virus vaccines are also known as conventional vaccines, including inactivated vaccines and live attenuated vaccines, among which live attenuated vaccines use artificial directional mutation methods, or screen out live microorganisms with highly weakened or basically non-toxic virulence from nature vaccines made. After inoculation of the live attenuated vaccine, it has a certain growth and reproduction ability in the body, and the immune effect is strong and long-lasting. Generally, it only needs to be vaccinated once, and the immunity will be maintained for life. In addition to stimul...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00C12N5/10C12R1/93
Inventor 王晓佳
Owner 王晓佳
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