Attenuated and adapted strain of pseudomonas for delivering antigens

Patent metadata

Field	Value
Publication number	US-9556442-B2
Application number	US-201214364322-A
Country	US
Kind code	B2
Filing date	Dec 12, 2012
Priority date	Dec 12, 2011
Publication date	Jan 31, 2017
Grant date	Jan 31, 2017

Abstract

A process for obtaining an adapted strain of Pseudomonas includes the following steps: deleting the genes ExoS, ExoT, aroA and IasI in an initial Pseudomonas strain cultivated in a LB medium; progressively cultivating this strain in a chemically defined medium based on a glucose minimal medium supplemented with magnesium and calcium; wherein the adapted strain presents the same toxicity and secretion capacities than the initial strain, and its doubling time when cultivated in the chemically defined medium is less than 60 minutes. An adapted strain is furthermore treated to become ‘killed but metabolically active’.

First claim

The invention claimed is:

1. An adapted strain of Pseudomonas , the adapted strain of Pseudomonas having deletions of ExoS, ExoT, aroA, and IasI genes and a doubling time when cultivated in a chemically defined medium comprising magnesium and calcium of less than 60 minutes, wherein the adapted strain of Pseudomonas has been deposited at the National Collection of Microorganisms Cultures (CNCM; Institut Pasteur) under the filing number I-4564.

2. A process for obtaining the adapted strain of Pseudomonas according to claim 1 , comprising:
deleting genes ExoS, ExoT, aroA and IasI in an initial Pseudomonas strain cultivated in a LB medium;
progressively changing the culture medium from LB medium to a chemically defined medium comprising magnesium and calcium; and
selecting the adapted strains presenting a doubling time inferior to 60 minutes when cultivated in the chemically defined medium.

3. The process according to claim 2 , wherein the chemically defined medium has the following composition: a glucose minimal medium M9 supplemented with 1 mM Mg 2+ and 1 mM Ca 2+ .

4. The process according to claim 2 , wherein the adapted strain is furthermore treated to become ‘killed but metabolically active’.

5. An immunogenic composition comprising the adapted strain of Pseudomonas according to claim 1 .

6. A composition for inducing an immune response in a host, the composition comprising the adapted strain of Pseudomonas according to claim 1 .

Assignees

• Univ Joseph Fourier-Grenoble 1
• Centre Hospitalier Univ Grenoble
• Univ Grenoble Alpes

Inventors

• Le Gouellec Audrey
• Toussaint Bertrand
• Polack Benoît

Classifications

• C12N1/20

  Bacteria; Culture media therefor · CPC title

• C12N15/102

  Mutagenizing nucleic acids · CPC title

• A61K2039/522

  avirulent or attenuated · CPC title

• A61K2039/523

  expressing foreign proteins · CPC title

• C12N1/36 Primary

  Adaptation or attenuation of cells · CPC title