Neutrophil modulation by IL-1β-induced fibroblasts

Neutrophils are key contributors to the pathogenesis of SLE. Using single-cell sequencing data from SLE skin lesions, we identified a subcluster of CXCL1+ fibroblasts primarily induced by IL-1β. To better understand the effects of CXCL1+ fibroblasts on neutrophils, we established a conditional culture model using culture supernatants from control and IL-1β-stimulated fibroblasts. Neutrophils were collected after 21 hours of conditional culture and subjected to RNA-seq. Our findings revealed that neutrophils exhibited a distinct pro-inflammatory phenotype, expressing higher levels of pro-inflammatory cytokines such as IL1B, TNFA, IL8, and OSM. Fibroblasts were treated with either control or IL-1β(10 ng/mL) and cultured for 48 hours. The fibroblast culture supernatants were collected and used for conditional culturing with neutrophils. Neutrophils were then collected, and RNA was extracted using Trizol and assessed for concentration and quality with a Nanodrop 2000 spectrophotometer. DESeq2 analysis was performed to identify significantly differentially expressed genes (DEGs) using Q value < 0.05 and fold change > 2 as thresholds.