Lactobacillus kefiranofaciens, microbial inoculum and application of lactobacillus kefiranofaciens

A kind of Lactobacillus and kumiss technology, applied in the field of microbial agents, to achieve the effects of promoting tumor immune microenvironment, slowing down tumor growth, and optimizing the structure of intestinal flora

Active Publication Date: 2022-03-11
TIANJIN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no relevant report about using Lactobacillus kumi for immune checkpoint therapy in the prior art

Method used

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  • Lactobacillus kefiranofaciens, microbial inoculum and application of lactobacillus kefiranofaciens
  • Lactobacillus kefiranofaciens, microbial inoculum and application of lactobacillus kefiranofaciens
  • Lactobacillus kefiranofaciens, microbial inoculum and application of lactobacillus kefiranofaciens

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preparation example Construction

[0028] In the present invention, the preparation method of the bacterial agent preferably includes the following steps: inoculate Lactobacillus kefir ZW18 in MRS medium for cultivation, cultivate to the end of the logarithmic phase, centrifuge to remove the supernatant, and collect the bacterial sludge . In the present invention, the temperature of the cultivation is preferably 37-38°C; the rotational speed of the centrifugation is preferably 5000-6000rpm; the temperature of the centrifugation is preferably 4°C; the time of the centrifugation is preferably 8-10min. In the present invention, after obtaining the bacterium slime, it is preferred to mix the bacterium slime and the cryoprotectant at a mass ratio of 2:5, and freeze-dry to obtain the bacteria powder; the cryoprotectant is preferably 12% m / v Skim milk powder. After obtaining the bacterial powder, the present invention preferably also includes resuspending the bacterial powder to obtain a bacterial agent; the reagent...

Embodiment 1

[0046] Lactobacillus kefir ZW18 was inoculated in MRS medium, cultivated at 37°C to the end of the logarithmic phase, centrifuged (6000rpm / min, 4°C, 8min) to remove the supernatant, and collected the sludge. Freeze-drying was carried out with 12% skimmed milk powder as a protective agent, and ZW18 bacterial powder was obtained after drying. Take an appropriate amount of Lactobacillus mukemi ZW18 powder, resuspend it with normal saline and adjust the final concentration to 2×10 8 CFU / mL, cultured in pH 2.0~3.0 medium for 4h, the number of viable bacteria was 5×10 6 More than CFU; cultivated in the environment of 0-0.3% bile salt for 3 hours, the number of viable bacteria is 5×10 6 More than CFU. Therefore, ZW18 has good acid and bile salt resistance properties. And it was verified in the mouse animal body, and the survival of ZW18 could be observed by fluorescence imaging method. The result is as figure 1 shown. figure 1 A is the gut of a non-fluorescent mouse, and B is t...

Embodiment 2

[0048] Take an appropriate amount of Lactobacillus mukemi ZW18 powder, resuspend it with normal saline and adjust the final concentration to 5×10 9 CFU / mL, the bacterial agent was prepared. The bacterial agent is the drug for preventing or improving tumor diseases and promoting immune checkpoint therapy against tumors according to the present invention.

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Abstract

The invention provides lactobacillus kefiranofaciens, a microbial agent and application of the lactobacillus kefiranofaciens, and belongs to the technical field of microbial agents. The lactobacillus kefiranofaciens ZW18 disclosed by the invention can regulate tumor immunity, stimulate CD8 cell infiltration and promote expression of interleukin-12 (IL-12) and interferon gamma (IFN-gamma), so that tumor growth is slowed down. Secondly, the lactobacillus kefiranofaciens ZW18 can be used for optimizing the intestinal flora structure and improving the micro-ecological environment of the intestinal tract, and the abundance of beneficial bacteria such as PrevoteraceaeNK3B31group, Parabacteroides and the like in the intestinal tract can be improved; the beneficial bacteria can be metabolized to generate substances such as short-chain fatty acid, and organism immunity is regulated through blood, so that tumor growth is slowed down. In addition, the lactobacillus kefiranofaciens ZW18 can enhance the anti-tumor effect of the immune checkpoint therapy and overcome the anti-tumor drug resistance of the immune checkpoint therapy.

Description

technical field [0001] The invention belongs to the technical field of microbial bacterial agents, and in particular relates to a strain of Lactobacillus kumis, microbial agents and applications thereof. Background technique [0002] In recent years, immune checkpoint inhibitors (immune checkpoint inhibitors, ICI) that target immune checkpoints have become promising drugs for the treatment of tumors. A large number of studies have confirmed the effectiveness of ICI in clinical practice, but there are still many patients receiving ICI treatment who have not benefited. Improving the clinical response rate of immunotherapy and finding immunotherapy biomarkers has become a top priority for the breakthrough of immunotherapy. Gut flora is one of the emerging immune biomarkers in recent years, and gut flora plays a decisive role in PD-1 immunotherapy. Transplanting the feces of patients who have curative effect on PD-1 into the antibiotic mouse model can restore the mice to regai...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23L33/135A61K35/747A61K45/06A61P1/00A61P35/00A61P37/02C12R1/225
CPCC12N1/20A23L33/135A61K35/747A61K45/06A61P1/00A61P35/00A61P37/02A23V2002/00A23V2400/11A23V2200/3204A23V2200/324A23V2200/308A61K2300/00
Inventor 王艳萍赵婧琦耿伟涛吕梦欣
Owner TIANJIN UNIV OF SCI & TECH
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