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HA-mRNA vaccine for preventing influenza A virus infection

A technology of influenza A virus and influenza virus, which is applied in the field of medicine, can solve the problems of long development time, strict requirements, unsuitable use of influenza emergency, etc., and achieve good humoral immunity and good protective effect

Pending Publication Date: 2022-03-08
ACADEMY OF MILITARY MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Most of the existing FM1 virus vaccines are traditional inactivated vaccines, which have excellent safety and effectiveness; however, the development time is relatively long, and the production of vaccines requires the use of live virus operations and relatively strict requirements, which are not suitable for emergency use of influenza

Method used

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  • HA-mRNA vaccine for preventing influenza A virus infection
  • HA-mRNA vaccine for preventing influenza A virus infection
  • HA-mRNA vaccine for preventing influenza A virus infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Design, preparation and in vitro expression detection of embodiment 1, HA-mRNA

[0029] 1. Design and preparation of HA-mRNA

[0030] 1. A template sequence for artificially synthesizing HA-mRNA, as shown in sequence 1 of the sequence listing.

[0031] In sequence 1 of the sequence listing, the 1st to 72nd from the 5' end is 5'-UTR, the 73rd to 1773rd is CDS, and the 1774th to 1990th is 3'-UTR.

[0032] The schematic diagram of the template of HA-mRNA is as follows figure 1 shown.

[0033] 2. Insert the template sequence artificially synthesized in step 1 between the NheI and XhoI sites of the pcDNA3.1(+) vector to obtain a recombinant plasmid (sequenced and verified).

[0034] 3. Perform plasmid extraction on the recombinant plasmid obtained in step 2.

[0035] 4. Use XbaI and PmeI to double digest the recombinant plasmid extracted in step 3, recover and purify the linearized vector fragment containing HA with a size of about 7319bp.

[0036] 5. Configure the reac...

Embodiment 2

[0102] Embodiment 2, the situation of antibody production after HAmRNA immunization mice and the preventive and protective effect to influenza A virus infection

[0103] 1. Solution preparation

[0104] 1. Sodium pentobarbital solution (prepared at a dose of 0.18mL / 20g)

[0105] Weigh 60 mg of pentobarbital sodium, and prepare 6 mL of pentobarbital sodium solution with a concentration of 10 mg / mL with physiological saline (aqueous NaCl solution with a concentration of 0.9%).

[0106] 2. Protamine solution

[0107] Weigh 1 mg of protamine (Sigma Company), prepare 1 mL of protamine solution with a concentration of 1 mg / mL with normal saline, and place it on ice for use.

[0108] 3. HA-mRNA+protamine vaccine complex

[0109] Prepare 80 μg HA-mRNA per mouse and operate on ice; the concentration of HA-mRNA stock solution is 17.65 μg / μL.

[0110] HA-mRNA+protamine vaccine complex: Take out the frozen HA-mRNA from the -70°C refrigerator, transfer 15.9 μL of HA-mRNA solution (280 μg...

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Abstract

The invention discloses an HA-mRNA (Hepatitis A-messenger Ribonucleic Acid) vaccine for preventing influenza A virus infection. The invention provides HA-mRNA, and the nucleotide sequence of the HA-mRNA is RNA coded by a sequence 1 in a sequence table. The invention provides an HA-mRNA vaccine for preventing influenza A virus infection, the expression level of a serum antibody in a mouse body inoculated with the HA-mRNA vaccine is detected by adopting an ELISA (enzyme-linked immuno sorbent assay) method, the mouse is attacked by using a lethal dose of virus after the last immunization, and the form, weight and survival rate conditions of the mouse are observed and recorded. Results show that the HA-mRNA vaccine can cause good humoral immunity in a mouse body and has a good protection effect. The invention is of great significance to research on influenza A virus infection vaccines.

Description

technical field [0001] The invention relates to the field of medicines, in particular to an HA-mRNA vaccine used for preventing influenza A virus infection. Background technique [0002] mRNA vaccines are a new subtype of vaccines that are developing rapidly. The current fields of application mainly include: ① Research on the prevention and treatment of infectious diseases, such as Pollard et al., used cationic lipid DOTAP / DOPE to encapsulate the mRNA of type I human immunodeficiency virus antigen Gag, and subcutaneously immunized mice to induce the expression of type I interferon Secretion, and antigen-specific, functional T cell response research on the prevention and treatment of infectious diseases. ②Research on the prevention and treatment of tumors. For example, Zhou et al. used artificially synthesized human melanoma-associated antigen gp100 mRNA and Japanese Sendai virus (Hemagglutinating virus of Japan, HVJ) liposomes, and then directly injected into the spleen of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/005G01N33/543G01N33/569G01N33/58A61K39/145A61P31/16
CPCC07K14/005G01N33/56983G01N33/543G01N33/581A61K39/12A61P31/16C12N2760/16122C12N2760/16134A61K2039/53A61K2039/55516
Inventor 杨静王升启王鑫李春华黄慧媛
Owner ACADEMY OF MILITARY MEDICAL SCI
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