Supplementary Figure 1: Oxygen consumption rates are increased in PPBL B cells

(a) Age distribution of basal OCR and basal ECAR in primary B cells from HCs (n = 15) and PAD patients (n = 14). (b) Basal OCR in primary B cells from female vs. male and CMV^– vs. CMV⁺ HCs (n = 13) and PAD patients (n = 14). (c) Representative cell surface phenotype of B cells (CD19⁺ lymphocytes) from HCs and PPBL patients stained with CD24, CD38, TACI, and CD27 antibodies. HCs (n=2) and PPBL patients (n=2) were independently tested once. (d) CFSE dilution histogram of sorted CD19⁺ IgD⁺CD27^–; IgD⁺CD27⁺ and PPBL B cells stimulated with IL-21 and CD40L or CPG for 5 days (upper panel) and summarized in bar diagram (lower graph). Representative of 3 independent experiments. (e) Gating strategy for sorting MZ-like B cells from healthy controls. CD19⁺ cells were discriminated based on SSC/FSC area (upper left) and doublets were gated out (upper right). MZ-like B cells are IgD⁺CD27⁺ cells (lower left). Sorted MZ-like B cells after FACS sorting (lower right). (f) Relative distribution of CD4⁺ and CD8⁺ T cell subsets from HCs (n=3) and a PPBL patients (n=3). Representative of 3 independent experiments. (g) Representative histograms of CFSE dilution assays of anti-CD3/anti-CD28 activated CD4⁺ and CD8⁺ T cells from a HC and a PPBL patient. Representative of 3 independent experiments. (h) T cell reactivity against CMV peptides (pp65 and IE-1) between HCs (n=2) and PPBL patient T cells (n=2) were assessed by IFN-gamma ELISpot. The result is positive if the geometric mean is > 10 (red line) (i) Representative mitochondrial perturbation profiles of primary T cells from HCs and PPBL patients. Summary graphs present basal OCR (left) and ECAR (middle) of primary T cells from HCs (n = 14) and PPBL patients (n = 3). Representative mitotracker red (MTR) and mitotracker deep red (MTDR) staining of primary B cells from HCs (n = 3) and PPBL patients (n = 3) (right). Representative of 3 independent experiments. Pooled data are presented as mean ± SEM. Statistical significance was assessed by two-sided unpaired t-test (b,h). * p < 0.05, ** p < 0.01, ns, not significant.