Transcriptomic profile of TLR7-induced DN2 B cell development with and without IL-4 Interference in vitro using B cells isolated from SLE patients.

The effects of IL-4 on B cell development under an in vitro condition to polarize B cells into the double-negative 2 lineage were investigated. Using single-cell RNA sequencing (scRNA-seq), we analyzed the transcriptomic profiles of B cells from systemic lupus erythematosus (SLE) patients treated with or without IL-4 at 4 hrs and 18 hrs following the stimulation. Our results revealed that IL-4 pre-treatment led to a decrease in the expression of interferon response genes and DN2 signature genes, while promoting the emergence of a B cell population characterized by DN4 signature genes. This has become evident at the 18 hr time point. Overall design: B cells isolated from SLE patients were stimulated in RPMI with 10% FBS supplemented with or without IL-4 for 1 h. Cells were then stimulated with a combination of B cell polarication cocktail that exhibited a high potency to polarize B cells into the T-bet+ IgD-CD27- phenotype. Cells were harvested at 4 and 18 hrs, followed by hash-tag antibody labeling and scRNA-seq analysis.