Method of culturing segmented filamentous bacteria in vitro

Patent metadata

Field	Value
Publication number	US-11725183-B2
Application number	US-201916548280-A
Country	US
Kind code	B2
Filing date	Aug 22, 2019
Priority date	Dec 23, 2014
Publication date	Aug 15, 2023
Grant date	Aug 15, 2023

Abstract

The present invention relates to an in vitro method of culturing a segmented filamentous bacterium strain, comprising co-culturing said segmented filamentous bacterium strain with a eukaryotic host cell, wherein the culture is performed at an O 2 level inferior to 5% in a rich tissue culture liquid medium containing bacterial medium components including iron. The present invention also relates to methods for genetically modifying a segmented filamentous bacterium strain comprising a step a culturing the strain in vitro.

First claim

The invention claimed is:

1. A genetically modified segmented filamentous bacteria (SFB) strain expressing a foreign antigen of interest obtained from a microorganism or a cell different from a SFB.

2. The genetically modified SFB strain according to claim 1 , wherein the SFB strain is in the form of a filament, an intracellular offspring or a spore.

3. The genetically modified SFB strain according to claim 1 , wherein said foreign antigen of interest is obtained from a bacterium, a virus or a fungus organism.

4. The genetically modified SFB strain according to claim 3 , wherein said foreign antigen of interest is obtained from mainly surface proteins of the diarrheal pathogens Shigella , enterotoxic Escherichia coli (ETEC) or attaching and effacing lesion (A/E)-inducing enteropathogenic E. coli (EPEC).

5. The genetically modified SFB strain according to claim 1 , wherein said foreign antigen of interest is obtained from a tumor cell.

6. The genetically modified SFB strain according to claim 1 , wherein said foreign antigen of interest elicits a Th17 response.

7. The genetically modified SFB strain according to claim 1 , wherein the genetically modified SFB strain is obtained by co-culturing a SFB strain with a eukaryotic host cell at an O 2 level of less than 5% in a rich tissue culture liquid medium comprising bacterial medium components and iron, and wherein the co-culture is performed until the SFB strain releases intracellular offspring or spores; and by genetically transforming the SFB strain by conjugation, electroporation or chemical transformation.

8. An immunogenic composition comprising the genetically modified SFB strain according to claim 1 .

9. A method of treatment of a disease caused by a microorganism in a subject in need thereof comprising administering to said subject an effective amount of the genetically modified SFB strain of claim 1 expressing a foreign antigen of interest obtained from said microorganism.

10. The method according to claim 9 , wherein said SFB strain is administered by oral ingestion or by parenteral administration.

11. A method of treatment of a cancer in a subject in need thereof comprising administering to said subject an effective amount of the genetically modified SFB strain of claim 1 expressing a foreign antigen of interest obtained from said cancer.

12. The method according to claim 11 , wherein said SFB strain is administered by mucosal routes or by parenteral administration.

Assignees

• Pasteur Institut
• Fond Imagine
• Assist Publique—Hopitaux De Paris
• Univ Paris Descartes
• Institut National De La Sante Et De La Rech Medicale In Serm
• Univ Paris Cite
• Inst Nat Sante Rech Med

Inventors

• Eberl Gerard
• Bikard David
• Schnupf Pamela
• Cerf Bensussan Nadine
• Gaboriau-Routhiau Valerie
• Sansonetti Philippe

Classifications

• C12N1/20 Primary

  Bacteria; Culture media therefor · CPC title

• C12N13/00

  Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves · CPC title

• C12N15/74

  Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora · CPC title

• C12N2502/23

  Gastro-intestinal tract cells · CPC title

• Y02A50/30

  Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change · CPC title