Isolation and characterization of macrophage adapted Salmonella Typhimurium persister clones

Lotte Jelsbak, Maja Marie Sørensen, Kasper Lind Lauridsen, Ole Skovgaard, Malene Nybo Othendal Nielsen

Publikation: KonferencebidragKonferenceabstrakt til konferenceForskning

Resumé

Cultured macrophages are frequently used to investigate Salmonella-host
interactions on a population scale. However, in recent years it has been
shown that sub-populations of intracellular Salmonella develop during
infection (Dr. Helaine and co-workers). Specifically, a high level of intracellular
persister bacteria was reported. These results show that Salmonella
undergo adaptive changes in response to the host-environment at a regulatory
single-cell level; however, the extent to which individual Salmonella
sub-populations adapt at a genetic level remains to be investigated.
Here we have investigated genetic changes of Salmonella in response to
brief exposures to the host environment, and in particular investigate genetic
adaptation within the persister population. Macrophages were infected
with Salmonella bacteria for 30 minutes. Persister bacteria were isolated and
cultured for a new round of infection. For every round we observed a gradual
increase in the number of persister-bacteria isolated from the macrophages.
After 7 rounds, we randomly picked 6 isolates and subjected these to
WGS. The six isolates were clonal harboring the same 5 mutations. One mutation
was an in-frame 15 bp deletion resulting in a 5 amino acid shortening
of a conserved protein. This deletion is also observed in genomes of clinical
Salmonella isolates.
Another mutation was a 161 bp deletion fusing two genes of an operon inframe
resulting in expression a larger protein. The novel fusion protein is
essential for infection and elicits increased persistence towards ampicillin.
In conclusion, during brief exposures to the host environment Salmonella
acquire distinct patho-adaptive mutations.
OriginalsprogEngelsk
Publikationsdato2018
StatusUdgivet - 2018
BegivenhedCopenhagen Bioscience: Averting the Post-antibiotic era - Challenges and developments - Favrholm Campus, Hillerød, Danmark
Varighed: 31 okt. 20183 nov. 2018
Konferencens nummer: 15
http://cph-bioscience.com/en/events/averting-post-antibiotic-era-challenges-developments

Konference

KonferenceCopenhagen Bioscience
Nummer15
LokationFavrholm Campus
LandDanmark
ByHillerød
Periode31/10/201803/11/2018
AndetAntibiotic resistance among bacterial pathogens is increasing posing serious health threats, and there is therefore a great need to face this challenge. The conference will bring together scientists across several research fields for discussions of possible solutions. It is clear that next generation therapeutic strategies will be rooted in creative multi-disciplinary research performed by the best scientists. Meet some of them at the forth-coming conference from 31 October to 3 November 2018.<br/><br/>Conference topics<br/><br/> Infection mechanisms, treatments and host response<br/> Antibiotic resistance<br/> Microbiomes, infections and resistance<br/> New antibiotics and treatments<br/>
Internetadresse

Citer dette

Jelsbak, L., Sørensen, M. M., Lind Lauridsen, K., Skovgaard, O., & Nybo Othendal Nielsen, M. (2018). Isolation and characterization of macrophage adapted Salmonella Typhimurium persister clones. Abstract fra Copenhagen Bioscience, Hillerød, Danmark.
Jelsbak, Lotte ; Sørensen, Maja Marie ; Lind Lauridsen, Kasper ; Skovgaard, Ole ; Nybo Othendal Nielsen, Malene. / Isolation and characterization of macrophage adapted Salmonella Typhimurium persister clones. Abstract fra Copenhagen Bioscience, Hillerød, Danmark.
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title = "Isolation and characterization of macrophage adapted Salmonella Typhimurium persister clones",
abstract = "Cultured macrophages are frequently used to investigate Salmonella-hostinteractions on a population scale. However, in recent years it has beenshown that sub-populations of intracellular Salmonella develop duringinfection (Dr. Helaine and co-workers). Specifically, a high level of intracellularpersister bacteria was reported. These results show that Salmonellaundergo adaptive changes in response to the host-environment at a regulatorysingle-cell level; however, the extent to which individual Salmonellasub-populations adapt at a genetic level remains to be investigated.Here we have investigated genetic changes of Salmonella in response tobrief exposures to the host environment, and in particular investigate geneticadaptation within the persister population. Macrophages were infectedwith Salmonella bacteria for 30 minutes. Persister bacteria were isolated andcultured for a new round of infection. For every round we observed a gradualincrease in the number of persister-bacteria isolated from the macrophages.After 7 rounds, we randomly picked 6 isolates and subjected these toWGS. The six isolates were clonal harboring the same 5 mutations. One mutationwas an in-frame 15 bp deletion resulting in a 5 amino acid shorteningof a conserved protein. This deletion is also observed in genomes of clinicalSalmonella isolates.Another mutation was a 161 bp deletion fusing two genes of an operon inframeresulting in expression a larger protein. The novel fusion protein isessential for infection and elicits increased persistence towards ampicillin.In conclusion, during brief exposures to the host environment Salmonellaacquire distinct patho-adaptive mutations.",
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year = "2018",
language = "English",
note = "Copenhagen Bioscience : Averting the Post-antibiotic era - Challenges and developments ; Conference date: 31-10-2018 Through 03-11-2018",
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Jelsbak, L, Sørensen, MM, Lind Lauridsen, K, Skovgaard, O & Nybo Othendal Nielsen, M 2018, 'Isolation and characterization of macrophage adapted Salmonella Typhimurium persister clones' Copenhagen Bioscience, Hillerød, Danmark, 31/10/2018 - 03/11/2018, .

Isolation and characterization of macrophage adapted Salmonella Typhimurium persister clones. / Jelsbak, Lotte; Sørensen, Maja Marie; Lind Lauridsen, Kasper; Skovgaard, Ole; Nybo Othendal Nielsen, Malene.

2018. Abstract fra Copenhagen Bioscience, Hillerød, Danmark.

Publikation: KonferencebidragKonferenceabstrakt til konferenceForskning

TY - ABST

T1 - Isolation and characterization of macrophage adapted Salmonella Typhimurium persister clones

AU - Jelsbak, Lotte

AU - Sørensen, Maja Marie

AU - Lind Lauridsen, Kasper

AU - Skovgaard, Ole

AU - Nybo Othendal Nielsen, Malene

PY - 2018

Y1 - 2018

N2 - Cultured macrophages are frequently used to investigate Salmonella-hostinteractions on a population scale. However, in recent years it has beenshown that sub-populations of intracellular Salmonella develop duringinfection (Dr. Helaine and co-workers). Specifically, a high level of intracellularpersister bacteria was reported. These results show that Salmonellaundergo adaptive changes in response to the host-environment at a regulatorysingle-cell level; however, the extent to which individual Salmonellasub-populations adapt at a genetic level remains to be investigated.Here we have investigated genetic changes of Salmonella in response tobrief exposures to the host environment, and in particular investigate geneticadaptation within the persister population. Macrophages were infectedwith Salmonella bacteria for 30 minutes. Persister bacteria were isolated andcultured for a new round of infection. For every round we observed a gradualincrease in the number of persister-bacteria isolated from the macrophages.After 7 rounds, we randomly picked 6 isolates and subjected these toWGS. The six isolates were clonal harboring the same 5 mutations. One mutationwas an in-frame 15 bp deletion resulting in a 5 amino acid shorteningof a conserved protein. This deletion is also observed in genomes of clinicalSalmonella isolates.Another mutation was a 161 bp deletion fusing two genes of an operon inframeresulting in expression a larger protein. The novel fusion protein isessential for infection and elicits increased persistence towards ampicillin.In conclusion, during brief exposures to the host environment Salmonellaacquire distinct patho-adaptive mutations.

AB - Cultured macrophages are frequently used to investigate Salmonella-hostinteractions on a population scale. However, in recent years it has beenshown that sub-populations of intracellular Salmonella develop duringinfection (Dr. Helaine and co-workers). Specifically, a high level of intracellularpersister bacteria was reported. These results show that Salmonellaundergo adaptive changes in response to the host-environment at a regulatorysingle-cell level; however, the extent to which individual Salmonellasub-populations adapt at a genetic level remains to be investigated.Here we have investigated genetic changes of Salmonella in response tobrief exposures to the host environment, and in particular investigate geneticadaptation within the persister population. Macrophages were infectedwith Salmonella bacteria for 30 minutes. Persister bacteria were isolated andcultured for a new round of infection. For every round we observed a gradualincrease in the number of persister-bacteria isolated from the macrophages.After 7 rounds, we randomly picked 6 isolates and subjected these toWGS. The six isolates were clonal harboring the same 5 mutations. One mutationwas an in-frame 15 bp deletion resulting in a 5 amino acid shorteningof a conserved protein. This deletion is also observed in genomes of clinicalSalmonella isolates.Another mutation was a 161 bp deletion fusing two genes of an operon inframeresulting in expression a larger protein. The novel fusion protein isessential for infection and elicits increased persistence towards ampicillin.In conclusion, during brief exposures to the host environment Salmonellaacquire distinct patho-adaptive mutations.

M3 - Conference abstract for conference

ER -

Jelsbak L, Sørensen MM, Lind Lauridsen K, Skovgaard O, Nybo Othendal Nielsen M. Isolation and characterization of macrophage adapted Salmonella Typhimurium persister clones. 2018. Abstract fra Copenhagen Bioscience, Hillerød, Danmark.