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Method for screening intestinal immunity suppression agents

An inhibitor, immune-related technology, applied in the field of immune biology, to achieve the effect of inhibiting inflammatory immune function

Active Publication Date: 2013-01-16
MEIJI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in recent years, issues such as the side effects of antibiotics or the emergence of multidrug resistance due to mutations have become a topic of discussion

Method used

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  • Method for screening intestinal immunity suppression agents
  • Method for screening intestinal immunity suppression agents
  • Method for screening intestinal immunity suppression agents

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0132] Example 1: Screening of anti-inflammatory immune organisms based on BIE cells

[0133] Using bovine intestinal epithelial (BIE) cells, 18 strains of lactic acid bacteria listed in Table 1 below were studied using enterotoxigenic Escherichia coli (ETEC 987P strain) as a pro-inflammatory cytokine. Each of them was heat-sterilized at 56° C. for 30 minutes.

[0134] Table 1 Table of test strains

[0135] strain

species

MEP221101

Lactobacillus reuteri

MEP221102

Lactobacillus reuteri

MEP221103

Lactobacillus casei

OLL2768

Lactobacillus casei

MEP221104

Lactobacillus casei

MEP221105

Lactobacillus casei

MEP221106

Lactobacillus casei

MEP221107

Lactobacillus casei

MEP221108

Lactobacillus casei

MEP221109

Lactobacillus casei

MEP221110

Lactobacillus rhamnosus

MEP221111

Lactobacillus rhamnosus

MEP221112

Lactobacillus rhamnosus

...

example 2

[0145] Example 2: Expression of Toll-like receptor signaling inhibitors based on immunobiology in BIE cells

[0146] 1.5 × 10 in a 24-well plate 4 BIE cells were inoculated per well and cultured for 3 days. On the third day of culture, triacylated lipopeptide Pam3CSK4 (200ng / ml) or OLL2768 strain (5×10 7 each / 1ml) stimulated for 12, 24, 36, 48 hours, and the Toll-like receptor signaling inhibitors (MKP-1 (MAPK phosphatase 1: MAPK phosphatase 1), IRAK-M (Interleukin-1 Receptor-Associated Kinase M: Interleukin-1 receptor-related kinase M), SIGIRR (Single Ig IL-1-Related Receptor: single IgIL-1-related receptor), BCL-3 (B-cell CLL / Lymphoma 3: B cell Expression of CLL / Lymphoma 3), Tollip (Toll-Interacting Protein: Toll Interacting Protein), A20 (TNFAIP3 ((Tumor Necrosis Factor, Alpha-Induced Protein 3: Tumor Necrosis Factor, Alpha-Induced Protein 3)).

[0147] In addition, after stimulating BIE cells with OLL2768 strain for 48 hours, the wells of the plate were washed three tim...

example 3

[0151] Example 3: Evaluation of anti-inflammatory activity in porcine intestinal epithelial (PIE) cells

[0152] Using PIE cells, the anti-inflammatory activity was evaluated in the same manner as in Example 1. That is, in a 12-well plate with 3 × 10 4 PIE cells were seeded per well at 37°C, 5% CO 2 conditions for 3 days. Add 500 μl of each test lactic acid bacteria to the wells other than the medium control and ETEC control at 100 MOI, at 37 °C, 5% CO 2 conditions for 48 hours. Wash the 12-well plate with PBS, add 500 μl of DMEM medium (10% FCS, 1% SP) to each well, and inoculate dead bacteria of ETEC (strain 987P) to 5.0×10 wells other than the medium control area. 7 cells / well (100MOI), stimulate PIE cells for 12 hours. After the stimulation, 500 µl of TRIzol was added to each well, and the cell lysate was recovered by pipetting. Total RNA was extracted, and the mRNA expression levels of IL-6, IL-8 and MCP-1 in each reagent were calculated by real-time PCR method, and...

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Abstract

Disclosed is a method that efficiently screens immunobiotics that can effectively suppress an inflammatory immunologic effect induced in the intestines. The present disclosures relate to: the method for screening suppression agents that suppress an inflammatory immunologic effect in intestinal epithelial cells; a bacterial strain isolated by means of said screening method; and a composition containing said bacterial strain.

Description

technical field [0001] The invention relates to a screening method for immunobiotics (Immunobiotics) capable of inhibiting inflammatory immunity in the intestine, a bacterial strain screened by the method, a new bacterial strain with the inhibitory effect and a composition with the bacterial strain. Background technique [0002] The discovery and use of Antibiotics is very important to the development of medicine today. However, in recent years, issues such as side effects of antibiotics or multidrug resistance due to mutations have become topics of discussion. Therefore, relative to "antibiotics", the idea of ​​"probiotics" is widely recognized in the world today. Probiotics refer to "beneficial microorganisms and their proliferation-promoting substances that can improve the bacterial flora in the digestive tract and bring beneficial effects to the host." The flora (flora of bacteria), the idea of ​​preventing and improving diseases while trying to make the flora healthy....

Claims

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Application Information

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IPC IPC(8): C12N1/20
CPCC12N1/20C12R1/245A23V2002/00A61K35/747A23L1/3014G01N2800/06G01N33/6893C12R1/225A61K2035/115G01N33/50A23L33/135A61P1/04A61P29/00A61P37/06C12R2001/245C12N1/205C12R2001/225A23V2200/32A23V2200/3204
Inventor 池上秀二牧野圣也木村胜纪北泽春树麻生久高梨直也细矢翔一
Owner MEIJI CO LTD
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