human monocytic cell line thp 1  (ATCC)

Journal: bioRxiv

Article Title: Bacterial extracellular vesicles indirectly destabilize a human stem cell–derived blood–brain barrier on-chip through pro-inflammatory stimulation of immune cells

doi: 10.64898/2026.01.23.701361

Figure Lengend Snippet: ( a ) Immunofluorescence images of BMECs in a multiwell plate stained for nuclei (blue) and ICAM-1 (magenta) following 16–17 h treatment with 50% hECSR medium + 50% conditioned medium from THP-1 macrophages. The THP-1s had been treated with or without 1×10 8 BEVs ml -1 for 24 h prior to collection of the conditioned medium. ( b ) Barplot of fluorescence intensity of ICAM-1 divided by cell count and normalized to the BMECs exposed to conditioned medium from untreated THP-1s. Performed unpaired two-tailed t-test. ( c ) Immunofluorescence images of BMEC/BPLC co-cultures immunostained for cell nuclei (blue) and claudin-5 (cyan) following 16–17 h treatment with 50% hECSR medium + 50% conditioned medium from THP-1 macrophages. Gaps in claudin-5 staining are circled with dotted lines (yellow). ( d ) Barplot of µSiM co-culture system permeability to lucifer yellow dye. Performed unpaired two-tailed t-test. ( e )–( k ) Cytokine analysis of conditioned medium from THP-1 macrophages that was collected following 24 h treatment with or without 1×10 6 BEVs ml -1 , 1×10 8 BEVs ml -1 , or 10 ng ml -1 LPS. Performed one-way ANOVA with Dunnett post-hoc test for each cytokine. ( e ) Barplot of TNF-α concentration. ( f ) Barplot of IL-1β concentration. ( g ) Barplot of IL-1RA concentration. ( h ) Barplot of IL-2 concentration. ( i ) Barplot of IL-8 concentration. ( j ) Barplot of GM-CSF concentration. ( k ) Barplot of MCP-1 concentration. Scale bars = 100 µm. Error bars = s.d. ✱ = p < 0.05, ✱✱ = p < 0.01, ✱✱✱ = p < 0.001, ✱✱✱✱ = p < 0.0001.

Article Snippet: Human leukemia monocytic type 1 (THP-1) cells (ATCC) were obtained from the lab of Dr. Karin Wuertz-Kozak.

Techniques: Immunofluorescence, Staining, Fluorescence, Cell Characterization, Two Tailed Test, Co-Culture Assay, Permeability, Concentration Assay

Journal: Frontiers in Immunology

Article Title: Exosome-mediated bidirectional immune dysregulation in tuberculosis: proteomic profiling reveals strain-specific strategies of virulent H37Rv and attenuated H37Ra

doi: 10.3389/fimmu.2025.1696299

Figure Lengend Snippet: Schematic of the experimental workflow. THP-1 cells differentiate into macrophages (via PMA) and are exposed to Mtb (uninfected/NC, avirulent H37Ra/RA, virulent H37Rv/RV). Macrophages undergo bacterial load/cell viability assays; exosomes undergo morphology/particle/marker protein characterization. Macrophages and released exosomes are analyzed via proteomics (extraction→trypsin digestion→iTRAQ labeling→HPLC separation→TripleTOF 6600 mass spec→bioinformatics). Host/bacterial proteins are analyzed for differentially expressed genes, enriched pathways and functions to explore virulence - dependent immune mechanisms.

Article Snippet: Human monocytic THP-1 cells (ATCC TIB-202) were maintained in RPMI-1640 medium (Gibco 11875093) supplemented with 10% fetal bovine serum (FBS, Gibco 10099141C), 1% penicillin-streptomycin (Gibco 15140122), and 50 μM β-mercaptoethanol (Gibco 21985023), with passaging every 3 days at a 1:5 ratio.

Techniques: Marker, Extraction, Labeling

Journal: Frontiers in Immunology

Article Title: Exosome-mediated bidirectional immune dysregulation in tuberculosis: proteomic profiling reveals strain-specific strategies of virulent H37Rv and attenuated H37Ra

doi: 10.3389/fimmu.2025.1696299

Figure Lengend Snippet: Heat map of functional proteins in RA/RV DEPs. (A) Virulence proteins in macrophages. (B) Antigenic proteins in macrophages. (C) Membrane associated proteins in macrophages. (D) Virulence proteins in exosomes. (E) Antigenic proteins in exosomes. (F) Membrane associated proteins in exosomes. Red bar represents up-regulation and blue bar represents down-regulation. The color key indicates the expression levels of the proteins. (G) Workflow of cytokines detection of NC-, RA- RV- exosome treated recipient THP-1 derived macrophages. (H) ELISA data showed exosomes from RA-infected donors triggered a significantly stronger pro-inflammatory response in recipient macrophages. Data are mean ± SD and representative of three independent experiments. ***, P < 0.001.

Article Snippet: Human monocytic THP-1 cells (ATCC TIB-202) were maintained in RPMI-1640 medium (Gibco 11875093) supplemented with 10% fetal bovine serum (FBS, Gibco 10099141C), 1% penicillin-streptomycin (Gibco 15140122), and 50 μM β-mercaptoethanol (Gibco 21985023), with passaging every 3 days at a 1:5 ratio.

Techniques: Functional Assay, Membrane, Expressing, Derivative Assay, Enzyme-linked Immunosorbent Assay, Infection