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
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