Fig 2: Complexing with SP-D, C1q or C3b enhance the interaction of PTX3 with dormant conidia of A. fumigatus.A Bar-graph representing the percentage of conidia positive for SP-D, C1q or C3b binding, as determined by flow-cytometry. B Bar-graph showing conidia positive for PTX3 binding alone or upon its complexing with SP-D, C1q or C3b (flow-cytometry). PTX3 binding was detected by anti-human PTX3-FITC. Median values are presented. Statistical analysis was performed with two-sided paired t-test (*p < 0.05). C Histograms (flow-cytometry data) showing the conidial binding pattern of PTX3, directly or after complexing with SP-D, C1q or C3b. Data acquired from one experiment is presented; therefore, the control and PTX3 counts were the same in the SP-D + PTX3, C1q + PTX3 and C3b + PTX3 panels, while the binding assay was repeated with two independent batches of A. fumigatus (biological replicates), each time in duplicates (technical replicates). Source data are provided as a Source data file.

Fig 3: Differences in the number, cell sources, and phenotype of circulating microparticles (MPs) from patients with systemic lupus erythematosus (SLE) and healthy controls (HCs). (A) Representative dot plots of the electronic noise, platelets, and MPs. The distribution of MP sizes is shown on the right and the ranking of MP sizes was estimated using standard-sized beads based on FSC-A and SSC-A. The data from a control sample are shown. (B) Frequency of MPs of different sizes (upper) and the number (lower) of total MPs in patients with inactive SLE (iSLE) (n = 28), patients with active SLE (aSLE) (n = 32), and HCs (n = 60). Frequency of circulating (C) MPs-CD41a +, MPs-CD45 +, MPs-Annexin V +, (D) MPs-CP +, MPs-DNA +, and (E) MPs-C1q + in patients with iSLE (n = 28), patients with aSLE (n = 32), and HCs (n = 60). Comparisons in MP sizes among the groups were performed using a two-way ANOVA and the Bonferroni’s post hoc test. Other comparisons among groups were performed using the Kruskal–Wallis test and Dunn’s post hoc test; *p ≤ 0.05, **p ≤ 0.01, and ***p ≤ 0.001.

Fig 4: CDC and C1q-binding activities of glycoengineered anti-CD20 mAbs. (a, b) CDC activities of glycoengineered anti-CD20 mAbs. Raji cells were incubated with 16% human serum and were serially diluted with glycoengineered anti-CD20 mAbs. (a) Percentages of cell lysis plotted against mAb concentrations. (b) CDC activity of 1 µg/ml of G1aF mAb and G1bF mAb mixtures in different ratios. (c) C1q binding of glycoengineered anti-CD20 mAbs. Raji cells were opsonized with anti-CD20 mAbs and incubated with human serum. The cells were stained with FITC-conjugated anti-C1q antibody and the C1q-binding level was analyzed by flow cytometer. Data are presented as mean ± SD (n = 3).

Fig 5: Circulating EVs of seropositive patients and HCs differ in number, cell sources, and phenotypes. Plasma EVs (A) count (above) and frequency of different sizes (below) in anti-CCP−RF−, anti-CCP+RF+/−, and anti-CCPhiRFhi RA patients and HCs; (B) Left (above), representative dot plot of EVs in FSC-A of an anti-CCP+RF+/− patient and a HC, with the intensity of Pacific blue beads of different sizes. Right (above), representative microphotographs of EVs from a HC and an anti-CCP+RF+/− patient (below) using STEM. (C) Left: frequency of circulating EVs from different cellular sources: platelets (CD41a), leukocytes (CD45), endothelium (CD205), erythrocytes (CD235a), and other sources (EV negative for the evaluated molecules) from anti-CCP−RF−, anti-CCP+RF+/−, and anti-CCPhiRFhi RA patients and HCs. Right: frequency of circulating EVs from different leukocyte sources in anti-CCP−RF−, anti-CCP+RF+/−, and anti-CCPhiRFhi RA patients and HCs. (D) Frequency of circulating EVs positive to IgG + IgM-, IgG-IgM+, IgG+IgM+, C1q+, HMGB1+, and CP+ from anti-CCP−RF−, anti-CCP+RF+/−, and anti-CCPhiRFhi RA patients and HCs. Comparisons among the study groups were made by performing the Kruskal–Wallis test and Dunn’s post-hoc test. (E) Collective t-SNE of EVs (1 × 105) derived from five samples from each group analyzed and plotted (n = 20). Every dot represents a single EV, and the color indicates ArcSinh5-transformed expression values for each given marker analyzed and calculated over EVs from all samples varying from blue for lower expression to red for higher expression.