Fig 2: SARS-CoV-2 spike-binding antibody responses and SARS-CoV-2 spike-specific memory B cells in blood following vaccination with BNT162b2.a, Total anti-spike IgG and subclasses after first and second doses of vaccine and in individuals with prior COVID-19. MFI, mean fluorescence intensity. b, Pearson’s correlation (r) between anti-spike IgG binding antibody responses after first dose and age (n = 134). c, Pearson’s correlation between anti-spike IgG (n = 134) binding antibody responses and neutralization by sera against SARS-CoV-2 in a spike lentiviral pseudotyping assay expressing wild-type spike (D614G). d, Anti-spike IgG subclass responses to first dose vaccine stratified by age (<80 and ≥80 years). e, CD19+ memory B cells (left, as percentage of PBMCs) and SARS-CoV-2 spike-specific CD19+IgG+IgM− memory B cells (right, as percentage of all memory B cells) from FACS-sorted PBMCs. n = 16 for ≥80 years, n = 16 for <80 years; stratified by neutralizing response after first dose, n = 8 in each category. MFI – mean fluorescence intensity. Mann–Whitney test was used for unpaired comparisons. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; NS, not significant. Scatter plots show linear correlation line bounded by 95% CI; β, slope/regression coefficient. Error bars, s.d.

Fig 3: Flow cytometry analysis of immune cells in human bone marrow shows type 2 diabetes (T2D) is associated with an increased abundance of CD4+ and CD8+ cells. Scooped bone marrow (BM) from femoral head leftovers of orthopedic surgery was obtained from patients previously diagnosed to have T2D and controls without T2D (ND). (A) Lymphocytes were gated based on SSC-A vs. FSC-A, and singlets were selected from the FSC-A vs. FSC-H dot plot. (B) Subsequently, dead cells were excluded with Zombie NIR™ Fixable Viability Kit. (C–G) Lymphocytes were gated based on SSC-A vs. FSC-A, and singlets were selected from CD45+ cells to identify population subsets according to the staining for CD3 (T lymphocytes), CD19 (B lymphocytes), and CD16/CD56 (NK cells). (H–J) Gating for CD4 and CD8 (H) Bar graphs showing the frequency of CD4+ cells (I) CD8+ cells (J). Reported frequencies are illustrative of a representative case for each group. Values are mean ± SEM, with each point representing an individual case.

Fig 4: Patients with SLE with neutralizing anti-IFN-Abs display normalized frequencies of peripheral blood B cell subsets(A–C) Representative contour plots and graphs shown for patients with SLE with neutralizing anti-IFN-Abs (n = 10) or non-neutralizing anti-IFN-Abs (n = 14), anti-IFN-Ab-negative patients with SLE (n = 41), and healthy individuals (n = 15). Ex vivo frequencies of (A) immature (Imm; CD24hiCD38hi) and mature (Mat; CD24intCD38int) B cells gated within the naive (CD27−IgD+) subset; (B) naive (N; CD27−IgD+ IgM+) unswitched memory (USM; CD27+ IgD+) and double-negative (DN; CD27−IgD−) B cells gated within the total CD19+ population; and (C) class-switched memory B cells (CSMs) and plasmablasts/plasma cells (PB/PC) gated within the CD27+IgD− subset. All values are given as the percentage of total CD19+ population (gating strategy in Figure S3A).(D) Representative contour plots and graphs show frequencies of IL-10+ B cells within the total CD19+ population following 72 h in vitro CpGC stimulation of PBMCs isolated from patients with SLE with neutralizing anti-IFN-Abs (n = 7) or non-neutralizing anti-IFN-Abs (n = 12), anti-IFN-Ab-negative patients with SLE (n = 16), and healthy individuals (n = 13).∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 by non-parametric Kruskal-Wallis test with Dunn’s multiple comparison. Error bars are shown as mean ± SEM. Data are representative of at least 3 independent experiments.