human monocyte like cell line  (ATCC)

Journal: Infection and Immunity

Article Title: Echinococcus granulosus antigen B acts as an LPS-scavenging lipoprotein in vitro, preventing TLR4-mediated activation of dendritic cells

doi: 10.1128/iai.00361-25

Figure Lengend Snippet: nEgAgB competed with LPS for binding to THP-1 macrophages, as hHDL does. THP-1 macrophages (0.2 × 10 6 ) were incubated for 30 min with nEgAgB, hHDL (both 1–200 μg/mL), OVA (200 µg/mL), or PBS EBAb (Veh control), followed by incubation with LPS-Alexa Fluor-488. Two preparations of nEgAgB were assessed: one purified including the ultracentrifugation step (nEgAgB UC+IA ) and one without this step (nEgAgB IA ). After washing, cell fluorescence was measured by flow cytometry. LPS binding to THP-1 macrophages is presented as the mean ± SD of data normalized to the LPS condition in the absence of competitive molecules (Veh control). The data correspond to three independent experiments with analytical duplicates. Significant differences compared to Veh are indicated with **, *** or **** (two-way ANOVA and Tukey’s test, P < 0.01, P < 0.001 or P < 0.0001, respectively). Note that no differences were observed among the nEgAgB UC+IA and nEgAgB IA ability to compete with LPS binding to THP-1 macrophages.

Article Snippet: The human monocyte-like cell line THP-1 (American Type Culture Collection, ATCC, USA) was cultured in RPMI 1640 medium containing HEPES 10 mM, sodium bicarbonate 1.5 g/L, sodium pyruvate 1 mM, glutamine 2 mM, antibiotic/antimycotic solution (Capricorn), and heat-inactivated fetal calf serum (FCS, Probiomont, Montevideo, Uruguay) 10% (vol/vol) at 37°C in a humidified atmosphere with 5% CO 2 (vol/vol), following ATCC recommendations.

Techniques: Binding Assay, Incubation, Control, Purification, Fluorescence, Flow Cytometry