Co-culturing with Streptococcus anginosus alters Staphylococcus aureus transcriptome when exposed to tonsillar cells

doi:10.21203/rs.3.rs-3252967/v1

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Co-culturing with Streptococcus anginosus alters Staphylococcus aureus transcriptome when exposed to tonsillar cells

https://doi.org/10.21203/rs.3.rs-3252967/v1

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Improved understanding of Staphylococcus aureus throat colonization in the presence of other co-existing microbes is important for mapping S. aureus adaptation to the human throat, and recurrence of infection. Here, we explore the responses triggered by the encounter between two common throat bacteria, S. aureus and Streptococcus anginosus, in the presence of human tonsillar epithelial cells, to identify genes in S. aureus that are important for colonization competition. We performed an in vitro co-culture experiment followed by RNA sequencing to identify important pathways and biological processes affected. A total of 332 and 279 significant differentially expressed genes were identified after 1 h and 3 h post-infection, respectively. S. aureus expression of genes encoding proteins involved in adhesion, immune evasion and iron regulation or homeostasis were altered during co-culturing with S. anginosus and host cells. These findings may be useful for development of interventions against S. aureus throat colonization and could be further investigated to decipher the detailed roles of the identified genes in the host immune response in the context of a throat commensal landscape.

co-culture

Staphylococcus aureus

human tonsil epithelial cells

throat colonization

transcriptomics

SupplementaryData1.xlsx
Data 1: Summary of trimming, and mapping efficacy against S. aureus and S. anginosus.
SupplementaryFigureS1.docx
Figure S1. Co-culturing of S. aureus and S. anginosus shows minor cytotoxicity to the tonsillar cell line. LDH release from tonsillar cells into the supernatant was measured following exposure with/without S. aureus and S. anginosus for 1 h and 3 h. The orange bar without dot represents the percentage (%) of LDH released by the tonsillar cell line in the absence of bacterial coculture (negative control) whereas dotted orange bar presents the LDH % release by the tonsillar cell line in the presence of bacterial co-culture. The bacterial cytotoxicity was calculated as a percentage of maximum LDH release control (positive control). The results are based on three independent experiments.
SupplementaryTableS1.xlsx
Table S1: Three hundred and thirty-two differentially expressed genes (DEGs) were identified in S. aureus co-cultured with S. anginosus with/without tonsillar cells after 1 h of exposure identified by RNA-seq. Out of 332 only 310 genes were pre-annotated and the remaining were unknown (NA). From known genes, 155 were identified as commonly expressed genes and 90 as uniquely differentially expressed genes at 1 h.
SupplementaryTableS2.xlsx
Table S2: Two hundred and seventy-nine differentially expressed genes (DEGs) were identified in S. aureus co-cultured with S. anginosus with/without tonsillar cells after 3 h of exposure identified by RNA-seq. Out of 279, only 207 genes were pre-annotated and the remaining were unknown (NA). From known genes, 155 were identified as commonly expressed genes and 52 as uniquely differentially expressed genes at 3 h.
SupplementaryTableS3.docx
Table S3: Gene ontology analysis of differentially expressed genes associated with S. aureus during coculturing with S. anginosus and tonsillar cells.
SupplementaryTableS4.xlsx
Table S4: Differentially expressed genes in S. aureus during co-culturing with S. anginosus compared to S. aureus monoculture, both in the absence of tonsillar cells after 1 h of exposure. A total of sixty-three genes were differentially expressed, of which 34 were upregulated and 29 were downregulated. Only 49 genes were annotated out of 63. Unknown genes were referred to as NA.
SupplementaryTableS5.xlsx
Table S5: Differentially expressed genes in S. aureus due to coculturing with S. anginosus compared to S. aureus monoculture, in the absence of tonsillar cells after 3 h. A total of sixty-three genes were differentially expressed, of which 39 were upregulated and 33 were downregulated. Only 49 genes were annotated out of 72. Unknown genes were referred to as NA.

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Co-culturing with Streptococcus anginosus alters Staphylococcus aureus transcriptome when exposed to tonsillar cells

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