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Weissella bacteria-derived nanovesicle and use thereof

A technology of Weissella and Weissella esophagus, applied to the composition for reducing or treating inflammatory diseases, the field of prevention

Pending Publication Date: 2021-07-30
MD HEALTHCARE INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there have been no reports on therapeutic techniques using extracellular vesicles derived from bacteria of the genus Weissella

Method used

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  • Weissella bacteria-derived nanovesicle and use thereof
  • Weissella bacteria-derived nanovesicle and use thereof
  • Weissella bacteria-derived nanovesicle and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1. Analysis of in vivo absorption, distribution and excretion patterns of bacteria and vesicles derived from bacteria

[0064] To assess whether bacteria and vesicles derived from bacteria are absorbed systemically through the gastrointestinal mucosa, experiments were performed as follows. A dose of 50 μg of each of fluorescently labeled bacteria and vesicles derived from bacteria in the stomach of mice was administered to the gastrointestinal tract, and after 0 minutes, 5 minutes, 3 hours, 6 hours and 12 hours Measure fluorescence. As a result of observing the whole image of the mouse, as Figure 1A As shown, bacteria were not taken up systemically, but vesicles derived from bacteria were absorbed systemically at 5 minutes after administration, and strong fluorescence was observed in the bladder at 3 hours after administration, making it possible to see that the vesicles were excreted. to the urinary tract. Furthermore, it can be seen that vesicles are pres...

Embodiment 2

[0066] Example 2. Evaluation of whether bacteria and vesicles derived from bacteria penetrate the protective membrane of the intestinal mucosa

[0067] To assess whether bacteria and bacteria-derived vesicles that penetrate the protective membrane of the intestinal mucosa penetrate into intestinal tissue after direct administration of bacteria and bacteria-derived vesicles to the intestine, the Penetration into the intestinal tissue behind the protective membrane of the intestinal mucosa. To assess the presence of bacteria and vesicles in the intestinal mucosa, antibodies against bacteria and vesicles were prepared, attached to green fluorescent protein (GFP) and used, and in combination with 4,6-diamidino-2-phenylindole ( DAPI) staining and observed under a microscope.

[0068] As a result, it was confirmed that bacteria cannot pass through the protective membrane of the intestinal mucosa, however, vesicles derived from bacteria permeate through the intestinal mucosa into ...

Embodiment 3

[0069] Example 3. Isolation of vesicles from Weissella esophagus culture solution

[0070] Based on the above examples, the Weissella angustiformis strain was cultured, then vesicles were isolated therefrom and the characteristics of the isolated vesicles were analyzed. First, the Weissella angustiformis strain was cultured in a de Man-Rogosa and Sharpe (MRS) medium in an incubator at 37°C until the absorbance became 1.0 to 1.5, and then in a Luria-Bertani (LB) medium Subculture. Subsequently, the culture supernatant containing the strain was recovered and centrifuged at 10,000×g and 4°C for 20 minutes, and then the strain was removed and filtered through a 0.22 μm filter.

[0071] The filtered supernatant was concentrated by microfiltration to a volume of less than or equal to 50 ml by using a MasterFlex pump system (Cole-Parmer, US) with a 100 kDa Pellicon 2 Cassette filter membrane (Merck Millipore, US), and filtered through a 0.22 μm filter. The concentrated supernatan...

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Abstract

The present invention pertains to a Weissella bacteria-derived vesicle and a use thereof. The present invention have experimentally confirmed that Weissella bacteria-derived vesicles efficiently inhibit inflammatory mediator secretion due to inflammation-inducing pathogenic vesicles, and it is thus expected that a Weissella bacteria-derived vesicle according to the present invention will be able to be usefully used for the purpose of developing a composition for preventing, alleviating, or treating inflammatory diseases.

Description

technical field [0001] The present invention relates to nanovesicles derived from bacteria of the genus Weissella and uses thereof, and more particularly, to a method for preventing, reducing or treating inflammation using nanovesicles derived from bacteria of the genus Weissella. Composition of diseases. [0002] This application claims priority and benefit to Korean Patent Applications 10-2018-0158621 and 10-2019-0132136 filed at the Korean Intellectual Property Office on December 10, 2018 and October 23, 2019, respectively, the specifications and accompanying All content disclosed in the figures is incorporated in this application. Background technique [0003] Since the beginning of the 21st century, acute infectious diseases, which were considered epidemic diseases in the past, have become less important, while inflammatory diseases accompanied by immune dysfunction caused by disharmony between humans and microbiomes have become Major diseases change disease patterns....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/74A61K9/00A23L33/135A61K8/99A61P29/00A61Q19/00
CPCA23L33/135A61K8/99A61P29/00A61Q19/00A61K8/14A61K2800/413A61K9/0053A61K35/741A61K35/74A61K9/007A23V2002/00A23V2200/324
Inventor 金润根
Owner MD HEALTHCARE INC
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