2023
DOI: 10.15698/mic2023.01.789
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Cellular cholesterol licenses Legionella pneumophila intracellular replication in macrophages

Abstract: Host membranes are inherently critical for niche homeostasis of vacuolar pathogens. Thus, intracellular bacteria frequently encode the capacity to regulate host lipogenesis as well as to modulate the lipid composition of host membranes. One membrane component that is often subverted by vacuolar bacteria is cholesterol – an abundant lipid that mammalian cells produce de novo at the endoplasmic reticulum (ER) or acquire exogenously from serum-derived lipoprotein carriers. Legionella pneumophila is an accidental … Show more

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Cited by 9 publications
(12 citation statements)
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“…1b-c). 79,80 The Lp growth kinetics with human U937 macrophages peaked at 48hrs for both JR32 and Lp01 and the curves were comparable regardless of serum availability (Fig. 1 a-b).…”
Section: Resultsmentioning
confidence: 86%
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“…1b-c). 79,80 The Lp growth kinetics with human U937 macrophages peaked at 48hrs for both JR32 and Lp01 and the curves were comparable regardless of serum availability (Fig. 1 a-b).…”
Section: Resultsmentioning
confidence: 86%
“…To avoid a pyroptotic cell death response triggered by cytosolic delivery of flagellin, 78 which can convolute data interpretation in infection-based growth assays, we used a flagellin clean deletion mutant. We engineered two distinct L. pneumophila Philadelphia-1 derived strains – Lp01 and JR32 – to bioluminescence by inserting the LuxR operon ( luxC luxD luxA luxB luxE ) from Photorhabdus luminescens on the bacterial chromosome under the constitutive IcmR promoter using two different genetic approaches 79,80 (SFig. 1a).…”
Section: Resultsmentioning
confidence: 99%
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“…Bioluminescence has emerged as a reliable approach for quantitative analysis of bacterial growth under various conditions, including in cellular infections (44, 45). Measurements of light output from bioluminescent bacteria allows growth kinetics analysis in a high-throughput manner with great resolution (41). Because light production requires ATP, bioluminescence also reflects the metabolic state and viability of the producers (46).…”
Section: Resultsmentioning
confidence: 99%
“…Inocula for infections were prepared in PBSG, RPMI or RPMI containing inactivated human serum (10%) and the number of bacteria was confirmed by plating dilutions from the inoculum on GCB agar. Legionella pneumophila serogroup 1 strain JR32 Δ flaA luxR was used in this study (41). The Legionella strain was grown on charcoal yeast extract (CYE) plates composed of 1% yeast extract, 1%N-(2-acetamido)-2-aminoethanesulphonic acid (ACES; pH 6.9), 3.3 mM l-cysteine, 0.33 mM Fe(NO3) 3 , 1.5% agar, 0.2% activated charcoal] (42).…”
Section: Methodsmentioning
confidence: 99%