Application of Akkermansia muciniphila in the preparation of probiotics for treating or preventing avian influenza virus infection

A technology of avian influenza virus and microecological preparations, which is applied in the direction of antiviral agents, medical raw materials derived from bacteria, and dispersion liquid delivery, which can solve problems such as changes in vaccine protection effects

Active Publication Date: 2021-05-14
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the current vaccination has a good effect on controlling H7N9 influenza, the protective effect of the vaccine is also changing with the high variability of influenza; when the vaccination is consistent with the subtype of the clinically circulating strain, ideal protection effect, once the strain mutates, it is difficult to have cross-protection

Method used

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  • Application of Akkermansia muciniphila in the preparation of probiotics for treating or preventing avian influenza virus infection
  • Application of Akkermansia muciniphila in the preparation of probiotics for treating or preventing avian influenza virus infection
  • Application of Akkermansia muciniphila in the preparation of probiotics for treating or preventing avian influenza virus infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The preparation method of the probiotic preparation of anti-avian influenza virus infection:

[0029] 1. Preparation of pasteurized Akkermansia muciniphila preparations

[0030] Akkermansia muciniphila ATCC BAA-835. The lyophilized powder of the strain was resuspended with sterile PBS, and an appropriate amount of bacterial solution was picked with an inoculation loop to be rejuvenated by streaking on the BD company's brain heart infusion (BHI) agar supplemented with 0.25% mucin (Sigma) Incubate at 37°C for 48h under oxygen condition. Wash the bacterial colony with 1mL PBS, collect the bacterial suspension; take 50 μL of the bacterial suspension and spread it evenly on the BHI agar containing 0.25% mucin, and cultivate it under anaerobic conditions at 37°C for 48 hours; Rinse the bacterial lawn, collect the bacterial suspension, centrifuge at 4°C and 5000rpm for 10 minutes, discard the supernatant, resuspend the collected Akkermansia muciniphila pellet in PBS, count o...

Embodiment 2

[0033] Effects of Akkermansia muciniphila ATCC BAA-835 on mortality and body weight of mice infected with H7N9 avian influenza virus

[0034] 1. Effects on Mortality and Body Weight of SPF Mice

[0035]The experimental animals were 8-week-old female C57BL / 6 specific pathogen-free (SPF) mice, a total of 30 were randomly divided into 3 groups, 10 in each group, and all animals were raised in a biosafety level 3 laboratory (ABSL3) . The first group is the control group fed with sterile PBS, 200ul per mouse; the second group is the group fed with live Akkermansia muciniphila (Akkermansia muciniphila) ATCC BAA-835 preparation, 200ul per mouse ; The third group was fed with pasteurized Akkermansia muciniphila (Akkermansia muciniphila) ATCC BAA-835 group, each mouse 200ul

[0036] Antibiotic treatment: Before the probiotic preparations were fed to the test mice, the mice were treated with a compound antibiotic solution, and the compound antibiotics were added to the sterile water t...

Embodiment 3

[0047] Effects of the inactivated probiotic preparation prepared in Example 1 on the influenza virus content in the lungs of mice infected with the H7N9 avian influenza virus, on the structure of the lungs of mice, and on the contents of cytokines in the lungs and blood.

[0048] The 36 SPF mice were randomly divided into 2 groups, 18 in each group. The first group was fed with sterile PBS control group, 200ul per mouse; the second group was fed with pasteurized mucin A For Akkermansia muciniphila ATCC BAA-835 group, 200ul per mouse;

[0049] As in Example 2, all SPF mice were raised in a biosafety level 3 laboratory (ABSL3), and underwent the same steps of antibiotic treatment, infusion of probiotic preparations and H7N9 influenza virus infection.

[0050] Lung tissue processing: Lung samples were dissected on day 0, day 3, and day 5 after infection, with 5 mice per group at each time point. Add 1ml PBS to each lung for homogenization, centrifuge at 8000rpm / min for 5min, and...

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Abstract

The invention relates to the field of biological preparations, and specifically discloses the application of Akkermansia muciniphila in the preparation of probiotics for the treatment or prevention of avian influenza virus infection; the pasteurized Akkermansia muciniphila of the present invention Bacteria intervened in mice, infected with H7N9 influenza virus, and evaluated the survival rate, body weight change, lung virus content, lung pathological changes, and lung cytokine levels of mice in each group. The results showed that pasteurized Akkermansia muciniphila could significantly improve the weight loss and lung tissue damage caused by H7N9 influenza virus infection, inhibit the proliferation of lung virus and significantly increase the survival rate of mice infected with H7N9 influenza virus. The present invention is characterized in that the probiotics are safe, have no toxic and side effects, and are convenient for storage compared with other live bacteria probiotics. For this reason, this invention provides important technical support for the development of novel preventive anti-influenza preparations.

Description

technical field [0001] The invention relates to the field of biological agents against influenza virus infection, in particular to the application of Akkermansia muciniphila in the preparation of probiotics for treating or preventing avian influenza virus infection. Background technique [0002] Influenza is a worldwide infectious viral disease caused by influenza virus, a single-stranded negative-sense RNA virus (Steinhauer and Skehel 2002), belonging to the Orthomyxoviridae family. Influenza viruses cause approximately 1 billion infections worldwide, resulting in annual and intermittent epidemics (Chen et al 2017). Also, it is highly variable and can continuously recombine to form new subtypes. [0003] The importance of the gut microbiota has been widely recognized, and changes in the gut microbiota not only affect local cellular functions and immune responses, but also play an important role in respiratory diseases (Marsland et al 2015). A growing body of research has ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K35/74A61P31/16
CPCA61K9/0095A61K35/74A61P31/16
Inventor 金梅林胡晓通张强杨丽王婷孙小美康超钟鸣
Owner HUAZHONG AGRI UNIV
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