Biology subjects

Steil, L.

Publications and source records attributed to Steil, L..

4 recordsLinked to original sources

bioRxiv2025

The S. aureus serine protease-like protein B (SplB) is a potent allergen causing eosinophilic airway inflammation in mice

Background: Asthma is associated with Staphylococcus aureus colonization. Two hypotheses were proposed to explain this phenomenon: (1) the allergic environment in asthma favors S. aureus colonization and (2) S. aureus colonization creates a pro-allergic environment. Since several S. aureus virulence factors, such as the serine protease-like protein (Spl) B, elicit a type 2 biased immune response, we asked whether the pathogen itself can cause asthma. Objective: Test the ability of recombinant SplB of S. aureus to sensitize mice and induce allergic airway inflammation (AAI). Methods: Mice were treated with repeated intratracheal inoculations of either catalytically active SplB or an inactive mutant. AAI was assessed by evaluating airway hypersensitivity, immune cell infiltration, cytokines, mucus production, fibrosis, and specific serum IgE. We compared the outcome between wild-type and gene-deficient C57BL/6J mice, including recombination-activating gene knockout mice (Rag2-/-), interleukin-33 knockout mice (Il33 -/-), and protease-activated receptor 2 knockout mice (F2rl1-/-). Results: Intratracheal exposure to SplB sensitized the mice and caused eosinophilic airway inflammation and hyperresponsiveness. The development of asthma required both the proteolytic activity of SplB and a functional adaptive immune system. The soluble protease sensor IL-33 was necessary for eosinophil tissue invasion, whereas the membrane-bound protease sensor PAR2 was not. Conclusion: The serine protease SplB of S. aureus is a potent allergen. Based on this finding we propose a third mechanism to explain the relationship between S. aureus colonization and asthma: S. aureus can release allergens, such as SplB, that sensitize individuals and lead to the development of asthma.

immunology↗

bioRxiv2025

Identification of novel human cellular substrates of Staphylococcus aureus serine protease SplB

Staphylococcus aureus colonizes up to one third of the human population yet retains the capacity to cause invasive, life-threatening infections. The growing prevalence of antimicrobial resistance further complicates treatment. A major contributor to the versatility of S. aureus is its broad repertoire of virulence factors, among which secreted proteases facilitate dissemination from colonization sites into deeper tissues. Twelve extracellular proteases are secreted, with the serine protease-like protein (Spl) family (SplA-SplF), encoded within a single operon, accounting for half of them. Despite this prominence, the pathophysiological roles and substrate specificities of the Spl proteases remain poorly understood. Here, we employed a direct protein-protein interaction approach to identify novel SplB substrates in human serum. We demonstrate that SplB cleaves three intermediate filament proteins, namely desmin, vimentin, and nestin, as well as heat shock protein {beta}1 and -enolase, which have not previously been recognized as targets of S. aureus proteases. Moreover, SplB was found to cleave native IgG, a feature otherwise described only for the glutamyl endopeptidase V8. These findings expand the host protein repertoire targeted by SplB and suggest broader roles for Spl proteases in immune evasion and tissue invasion.

immunology↗

bioRxiv2025

Integrated genomic and proteomic analysis of the mouse-adapted Staphylococcus aureus strain JSNZ

Mouse-adapted Staphylococcus aureus strains have become increasingly relevant in infection research thanks to their ability to better recapitulate clinical infection dynamics in mouse models. However, detailed characterisations required to establish a corresponding reference strain are still lacking. The mouse-adapted CC88 strain JSNZ appears to be an ideal candidate for a reference strain, because CC88 is widespread among laboratory mice and frequently employed in mouse colonisation and infection models. Moreover, JSNZ demonstrates high genetic transformability comparable to that of commonly used laboratory strains. Here, we present a comprehensive genomic and proteomic characterisation of JSNZ. Whole genome sequencing was performed using a combination of short and long reads. Proteomic profiling was conducted under standard laboratory conditions in TSB and RPMI during exponential and stationary growth using LC-MS/MS. The updated, closed genome sequence of JSNZ was integrated into AureoWiki for user-friendly access and direct comparison to long-established reference strains. Genome data revealed a deletion in the restriction endonuclease gene hsdR, likely explaining the observed efficient transformation while retaining DNA modification capabilities. This positions JSNZ as a hub for genetic modification of other CC88 isolates. Proteomic profiling of JSNZ indicated broad similarity to common S. aureus reference strains. However, a striking exception was the novel serine protease Jep, which constituted approximately 75% of the exoproteome in stationary TSB cultures. Overall, these findings affirm JSNZ as a robust and genetically tractable model strain for murine S. aureus infection research and contribute a valuable standardised resource to enhance experimental reproducibility and cross-study consistency in the field. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=119 SRC="FIGDIR/small/674026v1_ufig1.gif" ALT="Figure 1"> View larger version (42K): org.highwire.dtl.DTLVardef@130b864org.highwire.dtl.DTLVardef@199b1f2org.highwire.dtl.DTLVardef@171b6e0org.highwire.dtl.DTLVardef@9d371f_HPS_FORMAT_FIGEXP M_FIG C_FIG

microbiology↗

bioRxiv2025

The global proteome of Streptococcus pneumoniae EF3030 under nutrient-defined in vitro conditions

Streptococcus pneumoniae is a pathobiont that colonises the upper respiratory tract of humans without causing symptoms but can cause a range of life-threatening diseases including pneumonia, sepsis, and meningitis. It also causes less severe, non-invasive infections such as otitis media and sinusitis. This bacterium thrives in the nasopharynx, where nutrient availability is limited, and has adapted to this environment by developing mechanisms to survive host stress and regulate protein abundance. To study the molecular biology of S. pneumoniae under in vitro and infection-related conditions, a suitable cultivation medium is essential for reproducible experiments. In this study, we optimized a chemically-defined minimal medium that mimics the in vivo nutrient-limited condition and used it for proteome analysis. This optimized medium not only shortened the lag phase but also improved the growth of S. pneumoniae clinical isolates and other streptococcal species. We applied this medium to analyse the global proteome of the pneumococcal colonising strain EF3030, focusing on the transition from the early to late log phase. Our proteomic analysis revealed distinct patterns of protein abundance in different functional categories including metabolism, amino acid synthesis, natural competence, RNA synthesis, cell wall synthesis, protein degradation, and stress responses. Notably, choline-binding protein CbpD, competence factors ComGA and ComEA as well as proteins involved in processing internalized single-stranded DNA (ssDNA) such as Dpr and DprA were higher in abundance in the late log phase. This proteomic profiling provides valuable insights into the pathophysiology of strain S. pneumoniae EF3030 under defined nutrient conditions.

microbiology↗