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Anti-bacterial composition against Th1 cell-inducing bacteria

An antibacterial composition, the technology of the composition, applied in the directions of bacteria, antibacterial drugs, drug combinations, etc., can solve the problems of complex and detailed mechanisms that are not fully clear, and achieve the effect of proliferation or activation inhibition

Active Publication Date: 2020-03-17
KEIO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If the elucidation of the indigenous flora of the mucous membrane progresses, it is likely to contribute to the development of new disease countermeasures and treatments for various diseases, but due to its complexity, the detailed mechanism is not yet fully elucidated

Method used

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  • Anti-bacterial composition against Th1 cell-inducing bacteria
  • Anti-bacterial composition against Th1 cell-inducing bacteria
  • Anti-bacterial composition against Th1 cell-inducing bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0252]

[0253] Before SPF mice (wild-type C57BL / 6) were forced to ingest Th1 cell-inducing bacteria, the following antibiotics were administered through drinking water for 4 days. In addition, mice not administered with these antibiotics were also prepared.

[0254] Antibiotics: ampicillin (200mg / L), tylosin (500mg / L), metronidazole (500mg / L), spectinomycin (200mg / L), vancomycin (200mg / L).

[0255] Culture Kp2H7 strain or Ka11E12 strain as a Th1 cell-inducing bacterium in LB broth to the logarithmic phase, and 1~2×10 8 CFUs were used to vaccinate mice.

[0256] After forced inoculation with Th1 cell-inducing bacteria, faeces were recovered 1, 3, 7, 14 and 21 days later, from which DNA was extracted. Then, using these DNAs as templates, qPCR was performed using the following primers specific to each bacterial strain, and the intestinal colonization amount of each bacterial strain was evaluated.

[0257] Klebsiella (ompK36-3_F: 5'-GCGACCAGACCTACATGCGT-3' [SEQ ID NO: 148], ...

Embodiment 2

[0269]

[0270] like image 3 As shown, in the same manner as in Example 1, germ-free mice were inoculated with the Kp2H7 strain. Then, one week after the inoculation, a human stool sample collected from a healthy person (#K) was orally administered. In addition, 31 to 94 days after the oral administration, administration of ampicillin was continued in the same manner as in Example 1 (hereinafter, the mice treated in this way are also referred to as "control").

[0271] In addition, germ-free mice were treated in the same manner as the control (hereinafter, also referred to as The treated mice were referred to as "chloroform-treated group").

[0272] Furthermore, except that metronidazole was administered in the same manner as in Example 1 one day before the oral administration of the human feces sample, the germ-free mice were treated in the same manner as the control (hereinafter, after such treatment, The mice were referred to as "metronidazole-administered group").

...

Embodiment 3

[0276]

[0277] Preparation of stool samples

[0278] The stool (#K stool sample, #F stool sample and #I stool sample) provided by each healthy volunteer (#K, F and I) was diluted to 5% with glycerol PBS solution (final concentration of glycerol: 20% by volume). Double the weight, filtered through a filter with a diameter of 100 μm, and stored at -80°C as a stock solution. In addition, the healthy person volunteer #K in this example is the same person as the healthy person #K in Example 2.

[0279] Production of Kp2H7 single-bacteria colonized mice

[0280] C57BL / 6N Jcl gnotobiotic mice (manufactured by Nippon CLEA Co., Ltd., 4 to 8 weeks old) were kept in a vinyl isolation box (sterile isolation box) for breeding (manufactured by Aishiem Co., Ltd.; ICM-1B) with free access to drinking water. The feeding conditions were raised for more than 1 week to carry out environmental acclimatization.

[0281] The Kp2H7 strain was cultured with Schaedler's blood medium, LB mediu...

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Abstract

With the aim of providing an anti-bacterial composition against oral bacteria, and the like, that induce proliferation or activation of Th1 cells in the intestinal tract, it was discovered that bacteria that restrict colonisation of the intestinal tract, and the like, by the oral bacteria, and the like, are among the intestinal flora. Moreover, the intestinal bacteria that restrict intestinal colonisation, and the like, by the oral bacteria, and the like, were successfully isolated.

Description

technical field [0001] The present invention is based on the understanding of the network based on the mechanism of biological constancy maintenance, variation, and loopholes based on the innovative cutting-edge research and development support project of the National Research and Development Corporation Japan Medical Research and Development Agency (AMED), and the unit type in 2012. Acquired from the research results of a commissioned project in the "Technology Creation for Optimal Medical Care" research area (research and development project name: "Development of new therapeutic methods for intractable diseases based on the understanding of the characteristics of intestinal indigenous bacteria"). [0002] The present invention relates to an antibacterial composition against bacteria that induce proliferation or activation of Th1 cells in the intestinal tract (hereinafter also referred to as "Th1 cell-inducing bacteria"). In addition, the present invention relates to a pharma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/74A61P1/00A61P31/04A61P37/06C12N1/20C12Q1/02C12Q1/6883G01N33/50
CPCA61P1/00A61P31/04A61P37/06C12N1/20G01N33/56911C12Q1/04G01N2800/065G01N33/6863A61K35/741Y02A50/30A61K35/74C12Q1/02
Inventor 本田贤也新幸二成岛圣子须田互服部正平古市宗弘河口贵昭水户部惠子
Owner KEIO UNIV
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