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

Journal: Frontiers in Immunology

Article Title: Carbohydrate fatty acid monosulphate ester adjuvant enhances the immunogenicity of influenza antigens via TLR4/2-dependent mechanisms

doi: 10.3389/fimmu.2026.1787181

Figure Lengend Snippet: CMS adjuvant enhances antigen-specific T cell responses to H1N1 HA peptide pools in human PBMCs. (A) Scheme of the AIM assay setup. PBMCs from healthy donors were treated with H1N1 HA peptide pools alone or in combination with the adjuvant for 6 days and restimulated with peptide pools for 16 hrs in the presence of anti-CD40 and anti-CD28, followed by surface staining and flow cytometry analysis of AIMs. (B) Representative dot plots showing frequencies of CD154 + CD137 + CD4 + and CD137 + CD8 + T cells for the indicated treatments. (C) Summarized frequencies of HA-specific CD154 + CD137 + CD4 + and CD137 + CD8 + T cells from independent donors (n=9). Statistics are **P < 0.01 ; ns, not significant as determined by Mann-Whitney’s U test (C, D) . AIM, activation-induced marker; SD, standard deviation.

Article Snippet: The cells were restimulated with H1N1 peptide pools at a concentration of 1 μg/ml for 16 hrs in the presence of CD40 monoclonal antibody (1 μg/ml, clone: HB14; Miltenyi Biotec) and CD28 monoclonal antibody (1 μg/ml, clone: 37407; R&D Systems).

Techniques: Adjuvant, Staining, Flow Cytometry, Activation Assay, Marker, Standard Deviation

Journal: Frontiers in Immunology

Article Title: Carbohydrate fatty acid monosulphate ester adjuvant enhances the immunogenicity of influenza antigens via TLR4/2-dependent mechanisms

doi: 10.3389/fimmu.2026.1787181

Figure Lengend Snippet: CMS enhances the immunogenicity of H7N9 HA protein antigen on human DCs. Immature DCs were left alone or treated with H7N9 HA (10 μg/ml) antigen alone or antigen + CMS (125 µg/ml) for 48 h. After incubation, cells were subjected to surface phenotyping by FACS. (A, B ) Representative histograms and scatter plots representing the mean ± SD (n = 6 donors) values of expression (median fluorescence intensities, MFI) of CD80, CD86, CD40, HLA-DR, CD54, and CD274. (C) Cell-free supernatants were collected and analyzed for cytokines. Amount of secretion (mean ± SD, n = 12-13) of IL-1β, IL-6, IL-8, IL-10, IL-12p70 and TNF-α, (all in pg/ml) (n = 10–13 donors). Statistics are *P < 0.05; **P < 0.01; ****P < 0.0001 as determined by one-way ANOVA with Tukey’s multiple comparisons post-test (B, C) . SD, standard deviation.

Article Snippet: The cells were restimulated with H1N1 peptide pools at a concentration of 1 μg/ml for 16 hrs in the presence of CD40 monoclonal antibody (1 μg/ml, clone: HB14; Miltenyi Biotec) and CD28 monoclonal antibody (1 μg/ml, clone: 37407; R&D Systems).

Techniques: Immunopeptidomics, Incubation, Expressing, Fluorescence, Standard Deviation