Loss of IL10RA rewires epithelial immune responses to intracellular Staphylococcus aureus 1 small colony variants in bovine mammary epithelial cells

ABSTRACT: Staphylococcus aureus small colony variants (SCVs) are a persistence-adapted phenotype associated with chronic and recurrent bovine mastitis. The interleukin-10 receptor ⍺ (IL10RA) is a key regulator of anti-inflammatory signalling in the mammary gland, yet its role in governing transcriptional responses to intracellular staphylococcal challenge remains unknown. Using CRISPR/Cas9-generated IL10RA knockout (KO) MAC-T bovine mammary epithelial cells (MECs) and RNA sequencing, we compared transcriptional responses to infection with S. aureus SCV Heba3231 and its isogenic parental strain (PS). The wild-type (WT) MAC-T cells mounted fundamentally divergent responses to the PS (8 DEGs) and the SCV (461 DEGs), with no transcriptional overlap, indicating strain-specific engagement of distinct molecular programmes. SCV infection of WT cells up-regulated lipid and sterol biosynthetic pathways, and suppressed genes associated with epithelial barrier integrity, including S100A8, S100A9, TGM3 and KRTDAP. IL10RA disruption dramatically amplified transcriptional dysregulation. The IL10RA-KO cells infected with the PS yielded 619 DEGs (77.4-fold increase over WT-PS), while IL10RA-KO cells infected with SCV yielded 1,297 DEGs (2.8-fold increase over WT-SCV). A moderate core of IL10RA-regulated genes (~32–36%) was shared across infection conditions. IL10RA loss derepressed pro-inflammatory cytokine pathways (TNF, IL-17), enhanced pro-apoptotic programmes during SCV infection, and suppressed phagosomal maturation and lipid metabolic gene programmes. Novel candidate genes, including CD79B, WFDC2, AMN and LY6E, were identified as potential biomarkers of IL10RA-dependent mammary immune signalling. These findings establish IL10RA as a broad transcriptional homeostasis regulator in bovine mammary epithelial cells during intracellular staphylococcal infection.

ORGANISM(S): Bos taurus

PROVIDER: GSE334081 | GEO | 2026/06/08

REPOSITORIES: GEO