Stimulation strategy matters: A B-cell focused single-cell transcriptomic study on in vitro stimulated immune cells

IFN-{gamma}/R-848 and ODN2006 produce the largest strategy-specific transcriptional differences in in vitro stimulated B cells from healthy donors, across 8- and 30-hour incubations.

• Why it matters: Phenotypic similarity from flow cytometry may mask distinct underlying transcriptional states, limiting confidence when stimulation strategies are treated as interchangeable.
• What they did: Single-cell RNA-seq atlas from 4 healthy donors (~2,029 cells per sample; 81,160 total; 56.3% annotated as B cells) compared T-cell independent (ODN2006, IFN-{gamma}/R-848) versus T-cell dependent (anti-CD40, anti-CD3/CD28) stimulators at 8 and 30 hours.
• The result: Overlap in differential abundance, gene expression, and cell-cell interactions is linked to T-cell (in)dependent mechanisms, while IFN-{gamma}/R-848 and ODN2006 drive the largest differentially expressed gene sets and IFN-{gamma}/R-848 plus anti-CD3/CD28 induce most cell-cell interactions.