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- Title
Generation and Characterization of Stable Small Colony Variants of USA300 Staphylococcus aureus in RAW 264.7 Murine Macrophages.
- Authors
Bivona, Dalida; Bonomo, Carmelo; Colombini, Lorenzo; Bonacci, Paolo G.; Privitera, Grete F.; Caruso, Giuseppe; Caraci, Filippo; Santoro, Francesco; Musso, Nicolò; Bongiorno, Dafne; Iannelli, Francesco; Stefani, Stefania
- Abstract
Intracellular survival and immune evasion are typical features of staphylococcal infections. USA300 is a major clone of methicillin-resistant S. aureus (MRSA), a community- and hospital-acquired pathogen capable of disseminating throughout the body and evading the immune system. Carnosine is an endogenous dipeptide characterized by antioxidant and anti-inflammatory properties acting on the peripheral (macrophages) and tissue-resident (microglia) immune system. In this work, RAW 264.7 murine macrophages were infected with the USA300 ATCC BAA-1556 S. aureus strain and treated with 20 mM carnosine and/or 32 mg/L erythromycin. Stable small colony variant (SCV) formation on blood agar medium was obtained after 48 h of combined treatment. Whole genome sequencing of the BAA-1556 strain and its stable derivative SCVs when combining Illumina and nanopore technologies revealed three single nucleotide differences, including a nonsense mutation in the shikimate kinase gene aroK. Gene expression analysis showed a significant up-regulation of the uhpt and sdrE genes in the stable SCVs compared with the wild-type, likely involved in adaptation to the intracellular milieu.
- Subjects
STAPHYLOCOCCUS aureus; MACROPHAGES; WHOLE genome sequencing; STAPHYLOCOCCAL diseases; NONSENSE mutation
- Publication
Antibiotics (2079-6382), 2024, Vol 13, Issue 3, p264
- ISSN
2079-6382
- Publication type
Article
- DOI
10.3390/antibiotics13030264