Figure 1.

Interrogation of autoreactive B cell subsets through tetramer sorting and transcriptomic classification of single cells. (A) Schematic of the experimental workflow for the tetramer-based isolation, single cell-RNA-seq, and simultaneous immunoglobulin repertoire and transcriptomic analyses of B cells. (B) Representative flow cytometry gating approach for the identification of B cells producing RF or ACPA. (C) Comparison of RF and ACPA B cell subtype frequencies in peripheral blood between RA patients and healthy donors. A minimum of 15,000 B cells were analyzed for each sample. * = P<0.05 (D) B cell subtype distribution using the BCellNet classifier for the recovered 2,349 single cells from seven individuals. The ACPA⁺ and RF⁺ B cells from subject R01 produced poor quality yields during library preparation and were excluded from analysis. No ACPA⁺ cells were recovered from subject R04 and HD5 and R04, and no RF⁺ cells were recovered from subject R48 and HD5.