Salmonella typhimurium-infected caenorhabditis elegans for identifying inhibitors of infection
AssigneeThe General Hospital Corporation
InventorsFrederick Ausubel, Alejandro Aballay, Peter Yorgey
Life of this patent
Estimating the term…
Abstract
Disclosed are screening methods for identifying the interplay between environmental and host signals (e.g., host-dependent or host-independent signals) and physiological pathogenic pathways that control or regulate genes responsible for establishing a persistent infection, as in the colonization of the gut of the nematode.
Claims40
An isolated Caenorhabditis elegans nematode infected with an isolated Salmonella typhimurium bacterium.
(canceled)
(canceled)
A method of screening for a virulence factor that enables a pathogen to develop a persistent infection in a nematode, comprising the steps of:
- (a) exposing a nematode to a mutagenized pathogen;
- (b) determining whether said mutant pathogen persistently infects said nematode, a reduction of disease in said nematode relative to that caused by the non-mutagenized pathogen indicating a mutation in a virulence factor that enables said pathogen to develop a persistent infection in said nematode;
- and
- (c) using said mutation as a marker for identifying said virulence factor.
A method of screening for a compound that inhibits a S. typhimurium bacterium in a C. elegans nematode, comprising the steps of:
- (a) providing a C. elegans nematode infected with a S. typhimurium bacterium;
- (b) contacting said infected C. elegans nematode with a test compound;
- and
- (c) determining whether the test compound inhibits said S. typhimurium bacterium in said infected C. elegans nematode.
(canceled)
(canceled)
(canceled)
A method of screening for a virulence factor that enables a pathogen to develop a persistent infection in a nematode, comprising the steps of:
- (a) exposing a nematode to a mutagenized pathogen expressing a detectable marker;
- (b) determining whether said mutant pathogen persistently infects said nematode by measuring the level of detectable marker in said nematode, where a decrease of the marker in said nematode relative to that caused by the non-mutagenized pathogen indicates a mutation in a virulence factor that enables said pathogen to develop a persistent infection in said nematode;
- and
- (c) using said mutation as a marker for identifying said virulence factor.
A method of screening for a compound that inhibits a S. typhimurium bacterial infection in a C. elegans nematode, comprising the steps of:
- (a) providing a C. elegans nematode infected with a S. typhimurium bacterium expressing a detectable marker;
- (b) contacting said infected C. elegans nematode with a test compound;
- and
- (c) determining whether said S. typhimurium bacterium infects said C. elegans nematode by measuring the level of detectable marker in said nematode, where a decrease of the marker in said C. elegans nematode indicates that said test compound inhibits a S. typhimurium bacterial infection in the nematode.
A method of screening for a virulence factor that enables a pathogen to develop a persistent infection in a nematode, said method comprising:
- (a) exposing a nematode to a pathogen expressing a gene not normally expressed by said pathogen;
- (b) determining whether said pathogen persistently infects said nematode by measuring the level of detectable marker in said nematode, where a decrease or increase of the marker in the nematode relative to that caused by the non-mutagenized pathogen indicates a mutation in a virulence factor or a virulence factor gene that enables the pathogen to develop a persistent infection in the nematode;
- and
- (c) using gene expressed by said pathogen as a marker for identifying said virulence factor.
A method of screening for a virulence factor that enables a pathogen to develop a persistent infection in a nematode, said method comprising:
- (a) exposing a nematode to a pathogen, wherein said pathogen overexpresses a pathogen gene;
- (b) determining whether the pathogen persistently infects the nematode by measuring the level of detectable marker in the nematode, where a decrease or increase of the marker in the nematode relative to that caused by the non-mutagenized pathogen indicates a mutation in a virulence factor or a virulence factor gene that enables the pathogen to develop a persistent infection in the nematode;
- and
- (c) using the mutation or virulence factor gene as a marker for identifying the virulence factor.
A method of screening for a virulence factor that enables a pathogen to develop a persistent infection in a nematode, comprising:
- (a) exposing a nematode to a pathogen expressing a detectable marker, the pathogen being mutagenized;
- (b) determining whether the mutant or otherwise altered pathogen persistently infects the nematode by measuring the level of detectable marker in the nematode, where a decrease or increase of the marker in the nematode relative to that caused by the non-mutagenized pathogen indicates a mutation in a virulence factor or a virulence factor gene that enables the pathogen to develop a persistent infection in the nematode;
- and
- (c) using the mutation for identifying the virulence factor.
A method of screening for a virulence factor that enables a pathogen to develop a persistent infection in a nematode, comprising:
- (a) exposing a nematode to a pathogen expressing a detectable marker, the pathogen expressing a gene not normally expressed by the pathogen;
- (b) determining whether the mutant or otherwise altered pathogen persistently infects the nematode by measuring the level of detectable marker in the nematode, where a decrease or increase of the marker in the nematode relative to that caused by the non-mutagenized pathogen indicates a mutation in a virulence factor or a virulence factor gene that enables the pathogen to develop a persistent infection in the nematode;
- and
- (c) using the gene expressed by said pathogen as a marker for identifying the virulence factor.
A method of screening for a virulence factor that enables a pathogen to develop a persistent infection in a nematode, comprising:
- (a) exposing a nematode to a pathogen expressing a detectable marker, the pathogen overexpressing a pathogen gene;
- (b) determining whether the mutant or otherwise altered pathogen persistently infects the nematode by measuring the level of detectable marker in the nematode, where a decrease or increase of the marker in the nematode relative to that caused by the non-mutagenized pathogen indicates a mutation in a virulence factor or a virulence factor gene that enables the pathogen to develop a persistent infection in the nematode;
- and
- (c) using the gene expressed by said pathogen as a marker for identifying the virulence factor.
40-48. (canceled)
Dependencies are inferred from the claim text.