cd40 monoclonal antibody  (Miltenyi Biotec)

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

Journal: Pharmaceutics

Article Title: Leveraging the Immune Response from LIFE Biomaterial and Photon-Flash in Pre-Clinical Pancreatic Cancer Treatment

doi: 10.3390/pharmaceutics17101273

Figure Lengend Snippet: LIFE Biomaterial formulated with gadolinium-based nanoparticles in addition to titanium dioxide nanoparticles and loaded with an anti-CD40 antibody. CT and MRI contrasts were observed from the LIFE Biomaterial injection within the pancreas tissue of the cadaver, as indicated by the yellow arrows.

Article Snippet: The monoclonal antibody anti-mouse CD40 (FGK4.5/FGK45) was bought from BioXcell (Lebanon, NH, USA).

Techniques: Titanium Dioxide, Injection

Journal: Pharmaceutics

Article Title: Leveraging the Immune Response from LIFE Biomaterial and Photon-Flash in Pre-Clinical Pancreatic Cancer Treatment

doi: 10.3390/pharmaceutics17101273

Figure Lengend Snippet: ( a ) Multiplex immunofluorescence images representing the immune cells’ infiltration within the tumor microenvironment. The markers represent the immune cells present in the tumor microenvironment, such as CD8 (FITC) and CD4 (Cy5) T-cells, CD3 (Polaris 780), and CD45 (Cy3). ( b ) Plots derived from immunofluorescence image analysis. The sum fluorescence intensity was analyzed from each tissue sample and plotted to discriminate the amount of immune cell infiltration according to the treatment administered compared to the control cohort. Higher CD4 T-cells infiltrations were only observed for the LIFE Biomaterial_20 µg-Anti-CD40 (*, p < 0.05) or those combined with Conv_20 Gy (*, p < 0.05) associated with the control group. Subsequently, higher CD8 cytotoxic T-cells (*, p < 0.05) were observed in the LIFE Biomaterial_Conv-10 Gy_20 µg-Anti-CD40-treated tumors compared to the no-treatment group.

Article Snippet: The monoclonal antibody anti-mouse CD40 (FGK4.5/FGK45) was bought from BioXcell (Lebanon, NH, USA).

Techniques: Multiplex Assay, Immunofluorescence, Derivative Assay, Fluorescence, Control

Journal: PLOS Pathogens

Article Title: Germinal Center B cells provide essential IL-1β signals to TFH cells via canonical NLRP3 inflammasome activity post influenza infection

doi: 10.1371/journal.ppat.1013404

Figure Lengend Snippet: Splenic B cells were enriched from naïve or A/PR8 infected mice at 15 dpi and stimulated with a combination of rBAFF, rIL2, R848, anti-CD40, rIL21, anti-IgM to examine intracellular (IC) IL-1β. Cells were gated on live, singlet lymphocytes to examine ( A ) B cells expressing IC IL-1β in each stimulation group. IL-1β expression in B cells was quantified by ( B ) MFI and by ( C ) frequency of IL-1β + B cells. ( A-C, ) Symbols representing each stimulation match their respective histogram and bar on the graphs. mLN from A/PR8 infected C57BL/6J mice were gated on live, singlet lymphocytes to determine ( D ) B cell populations including naïve, GC, and non-GC (nGC) B cells. ( E ) B cell populations were examined for IC IL-1β at 5,7,10,14 days post infection. ( F ) IC IL-1β was compared between naïve, GC and nGC B cells to determine the ( G ) frequency of IL-1β+ cells in each respective population and ( H ) quantify the MFI of IL-1β in each respective B cell population ( and ). Data are representative of 3 experiments with technical triplicates per stimulation group ( A-C ) and 3 experiments with 3 mice and graphs show individual points and mean ± SD ( C-G ). *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

Article Snippet: Mouse cells were stimulated with 10u/mL mouse IL-2 (MilliporeSigma, Rockville, MD), 5ng/mL recombinant BAFF (BioLegend, San Diego, CA),10ug/mL anti-mouse CD40 (BioXcell, Lebanon,NH), 10ng/mL recombinant IL-21 (Peprotech, Cranbury, NJ), 10ug/mL Goat anti-mouse IgM (Jackson ImmunoResearch, West Grove, PA) or 5ug/mL R848/Resiquimod (MilliporeSigma, Rockville, MD).

Techniques: Infection, Expressing

Journal: PLOS Pathogens

Article Title: Germinal Center B cells provide essential IL-1β signals to TFH cells via canonical NLRP3 inflammasome activity post influenza infection

doi: 10.1371/journal.ppat.1013404

Figure Lengend Snippet: B cells enriched from human tonsils were stimulated with a combination of rBAFF, rIL2, rIFNg, R848, anti-CD40, rIL21, anti-IgM, and anti-IgG to examine IC IL-1β. Cells were gated on live, singlet lymphocytes to examine B cells expressing IC IL-1β . ( A ) IL-1β was quantified by MFI and frequency f IL-1β + B cells. Symbols representing each stimulation match their respective histogram and bar on the graphs. Human tonsils were gated on live, single lymphocytes to examine naïve (CD19 + CD3-CD38-IgD+) and GC/active (CD19 + CD3-CD38 + IgD-) and intracellular IL-1β, caspase, and NLRP3 . Naïve and GC B cells, from human palatine tonsils, were compared for ( B ) IL-1β expression, ( C ) active caspase-1, and ( D ) coexpression of IL-1β and active caspase-1. Tonsil naïve and GC B cells were examined for ( E ) intracellular NLRP3 expression and ( F ) coexpression of NLRP3 and active caspase-1. ( A ) Data are representative of 4 experiments with technical triplicates per stimulation group. ( B-F ) Data are representative of 2 experiments with 4-5 patient samples per experiment. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

Article Snippet: Mouse cells were stimulated with 10u/mL mouse IL-2 (MilliporeSigma, Rockville, MD), 5ng/mL recombinant BAFF (BioLegend, San Diego, CA),10ug/mL anti-mouse CD40 (BioXcell, Lebanon,NH), 10ng/mL recombinant IL-21 (Peprotech, Cranbury, NJ), 10ug/mL Goat anti-mouse IgM (Jackson ImmunoResearch, West Grove, PA) or 5ug/mL R848/Resiquimod (MilliporeSigma, Rockville, MD).

Techniques: Expressing