Journal: Frontiers in Immunology

Article Title: Staphylococcus aureus serine protease-like protein B elicits a type 1/type 2 immune response in atopic dermatitis patients

doi: 10.3389/fimmu.2026.1798583

Figure Lengend Snippet: CD4 + T cells in the circulation react to SplB. PBMCs were isolated from healthy donors and AD patients. PBMCs (1 × 10 6 ) were stimulated with SplB in the presence of anti-human CD40 and anti-human CD28 for 24 h. (A) Expression of CD154 on CD4 + T cells with or without stimulation with SplB gated on live CD4 + T cells. (B) SplB-reactive, CD154 + T cells per 1 × 10 5 CD4 + T cells in healthy adults (n = 13, blue) and AD patients (n = 27, yellow); CD154 + T cell frequencies in unstimulated control cells were subtracted. P-values are based on Mann–Whitney U test. Median with interquartile range are depicted. ( C–F ) CD154 + CD4 + T cells were characterized by surface expression of CCR4, CCR10, CCR6, and CXCR3 to define (C) CD154 + Th1 (CXCR3 + CCR4 − CCR10 − CCR6 − ), (D) CD154 + Th2 (CXCR3 − CCR4 + CCR10 − CCR6 − ), (E) CD154 + Th17 (CXCR3 − CCR4 + CCR10 − CCR6 + ), and (F) CD154 + Th22 (CXCR3 − CCR4 + CCR10 + CCR6 + ) cells. Frequencies within 1 × 10 5 CD4 T cells are shown. Friedman test with Dunn’s multiple comparison test. (G) Th2/Th1 and (H) Th2/Th17 ratio of SplB reactive CD4 + T cells in healthy individuals and AD patients with mild (SCORAD <25), moderate (SCORAD 25–50) and severe disease (SCORAD >50). Each data point reflects a single patient. Kruskal–Wallis test with Dunn’s multiple comparison test. TT, tetanus toxoid, SEB, S. aureus enterotoxin (B) Median with interquartile range are depicted.

Article Snippet: To prevent CD154 internalization and provide co-stimulation and a survival signal, agonistic antibodies against CD40 (1 μg/mL, 130-094-133, clone HB14, Miltenyi, Bergisch Gladbach, Germany) and CD28 (0.5 μg/mL, 555726, clone CD28.2, Becton Dickinson GmbH, Franklin Lakes, NJ, USA) were added for 24 h. To investigate SplB-specific T cells, PBMCs were stained with antibodies (unless otherwise stated: from BioLegend, San Diego, CA, USA): PerCP-conjugated anti-human CD4 (317432, clone OKT4) or PE-Cy5-conjugated anti-human CD4 (555348, clone RPA-T4, Becton Dickinson GmbH, Franklin Lakes, NJ, USA) BV510-conjugated anti-human CD14 (301842, clone M5E2), BV510-conjugated anti-human CD19 (302242; clone HIB19), FITC-conjugated anti-human CD154 (310804; clone 24-31), APC-conjugated anti-human CXCR3 (FAB160A, clone 49801, R&D Systems Inc, Minneapolis, MN, USA), BV421-conjugated anti-human CCR4 (359414, clone L291H4), PE-Cy7-conjugated anti-human CCR6 (353418, clone G034E3), and PE-conjugated anti-human CCR10 (130-120-407, clone: REA326, Miltenyi, Bergisch Gladbach, Germany) ( ).

Techniques: Isolation, Expressing, Control, MANN-WHITNEY, Comparison

Journal: Science Advances

Article Title: Inflammatory arthritis irAE may represent a unique autoimmune disease primarily driven by T cells but likely not autoantibodies

doi: 10.1126/sciadv.aea4262

Figure Lengend Snippet: ( A to C ) Human naïve B cells were isolated from PBMCs and cultured under indicated conditions for 7 days; pembrolizumab or isotype control IgG4 (IgG4 Iso) was added on day 2; n = 8. (A) Expression of CD38 and CD27 on B cells. Right: Percentages of CD27 + CD38 − and CD27 + CD38 + B cells. (B) Expression of CD138 on CD27 + CD38 + B cells. Right: Percentage of CD138 + CD27 + CD38 + B cells. (C) Different immunoglobulin isotypes were measured in the culture supernatant of (A) and (B) by multiplex assay. ( D to F ) B cells from Humanized PD-1 (HuPD-1) mice were isolated and cultured with LPS, IL-4, BAFF, or ODN 2006, anti-IgM, IL-21, IL-4 or R848, anti-IgM, anti-CD40, IL-21, and IFN-γ for 3 days; pembrolizumab or isotype control was added on day 1. (D) Expression of IgG2c on activated B cells. FSC-H, Forward scatter height. Right: Percentage of IgG2c + B cells; n = 5. (E) Expression of IgG1 on activated B cells. Right: Percentage of IgG1 + B cells; n = 5. (F) Different immunoglobulin isotypes in the supernatant were measured by multiplex assay; n = 5. Data in graphs represent mean ± SEM. Significance was tested by two-way ANOVA.

Article Snippet: Condition 1: anti–human Ig (M + G + A) (2.5 μg/ml; Jackson ImmunoResearch, catalog no. 109-006-064), CpG oligodeoxynucleotide (ODN) (2.5 μg/ml; Invivogen, catalog no. tlrl-2006-1), anti–human CD40 (10 μg/ml; Bio X Cell, catalog no. BE0189), rhIL-21 (20 ng/ml; Peprotech, catalog no. 200-21-50UG), rhIL-4 (10 ng/ml; BioLegend, catalog no. 574004), and rhIL-2 (10 ng/ml; Peprotech, catalog no. 200-02-250UG).

Techniques: Isolation, Cell Culture, Control, Expressing, Multiplex Assay

Journal: Science Advances

Article Title: Inflammatory arthritis irAE may represent a unique autoimmune disease primarily driven by T cells but likely not autoantibodies

doi: 10.1126/sciadv.aea4262

Figure Lengend Snippet: ( A to F ) Bead-based multiplex assays were used to measure plasma concentration of IL-6, and IL-12p70 (A); TNF-α, IFN-γ, and IL-1β [B; HC ( n = 19), irAE ( n = 34), RAC ( n = 45), and ICI ( n = 9)]; IP-10 (CXCL10), CXCL11, and CXCL9 (C; HC, n = 17; irAE, n = 33; RAC, n = 46; ICI, n = 17); CCL20 (D); CX3CL1 (E); and CCL2 (F). ( G to J ) Human naïve B cells were isolated and cultured with anti–human CD40 (0.5 μg/ml), anti–human Ig (M + G + A) (2.5 μg/ml), and rhIL-21 (20 ng/ml) with IFN-α (100 ng/ml), IL-6 (100 ng/ml), IL-12 (100 ng/ml), control, or the combination of IFN-α, IL-6, and IL-12 for 7 days. Cells and culture supernatants were analyzed. (G) Representative flow plot of CD38 and CD138 expression on CD27 hi CD38 hi ASCs. Right: A summary of the percentage of CD138 + ASCs; n = 6. (H) Expression of CD11c and CD27 on CD27 − IgD − B cells. Right: Percentage of CD11c + IgD − CD27 − B cells; n = 6. (I) Expression of active-caspase-3 in B cells. Right: Percentage of active-caspase-3 + B cells from different groups; n = 3. (J) Different immunoglobulin isotype levels in the culture supernatants from (G) to (H) were measured by the multiplex assay; n = 6. Data in graphs represent mean ± SEM. Significance was tested by one-way ANOVA [(A) to (I)] and paired Student’s t test (J). [(A) to (F)] ICI, ICI control.

Article Snippet: Condition 1: anti–human Ig (M + G + A) (2.5 μg/ml; Jackson ImmunoResearch, catalog no. 109-006-064), CpG oligodeoxynucleotide (ODN) (2.5 μg/ml; Invivogen, catalog no. tlrl-2006-1), anti–human CD40 (10 μg/ml; Bio X Cell, catalog no. BE0189), rhIL-21 (20 ng/ml; Peprotech, catalog no. 200-21-50UG), rhIL-4 (10 ng/ml; BioLegend, catalog no. 574004), and rhIL-2 (10 ng/ml; Peprotech, catalog no. 200-02-250UG).

Techniques: Multiplex Assay, Clinical Proteomics, Concentration Assay, Isolation, Cell Culture, Control, Expressing