cell culture thp 1 cells  (ATCC)

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Filifactor alocis FtxA blocks inflammation and apoptosis pathways in monocytic cells

doi: 10.3389/fcimb.2026.1745721

Figure Lengend Snippet: Kinetic analysis of potential FtxA-dependent THP-1 cell toxicity over time (hours). (A) THP-1 cell toxicity in response to EVs of ATCC 35896 ( ftxA + ) or 148B-17U ( ftxA - ), as stimulated with whole cell extracts (MOI 200) obtained from the same F. alocis strains, respectively, and with recombinant FtxA. (B) Monitored THP-1 toxicity upon increasing concentrations of recombinant FtxA. Cell toxicity is expressed as the percentage of propidium iodide (PI)-positive cells, minus background readouts (untreated THP-1 cells; Control). Triton X-100 (0.1%) was used as a positive control for cell lysis. Shown are the means from three independent experiments. For each time point, P<0.001 between individual readouts for experimental treatments.

Article Snippet: To further validate the immunosuppressive effects of FtxA, a multiplex cytokine array assay was conducted on THP-1 cell culture supernatants after exposure to purified FtxA, and EVs obtained from ATCC 35896 ( ftxA + ), and 148B-17U ( ftxA − ).

Techniques: Recombinant, Control, Positive Control, Lysis

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Filifactor alocis FtxA blocks inflammation and apoptosis pathways in monocytic cells

doi: 10.3389/fcimb.2026.1745721

Figure Lengend Snippet: FtxA suppresses the immune response. THP-1 cells were exposed to F. alocis ftxA + and ftxA − EVs (3.5 μg/ml) and recombinant FtxA (1 μg/ml), respectively, for 4 h, followed by RNA-Seq analysis of two 4-h replicates. THP-1 cells treated with vehicle (PBS) were used as a control. Volcano plot indicating differential gene expression in THP1 cells treated with (A) ftxA + EVs and (B) ftxA − EVs. Top 20 differentially expressed genes are highlighted. (C) Venn diagram showing the distribution of differentially expressed genes in THP1 cells exposed to EVs ( ftxA + and ftxA − ), or recombinant FtxA. (D) Heatmap displaying differential gene expression in THP1 cells exposed to EVs ( ftxA + and ftxA − ), or recombinant FtxA. The genes involved in the immune response are indicated in the heatmap and are upregulated in the cells treated with ftxA − EVs and downregulated upon exposure to either ftxA − EVs or recombinant FtxA.

Article Snippet: To further validate the immunosuppressive effects of FtxA, a multiplex cytokine array assay was conducted on THP-1 cell culture supernatants after exposure to purified FtxA, and EVs obtained from ATCC 35896 ( ftxA + ), and 148B-17U ( ftxA − ).

Techniques: Recombinant, RNA Sequencing, Control, Gene Expression

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Filifactor alocis FtxA blocks inflammation and apoptosis pathways in monocytic cells

doi: 10.3389/fcimb.2026.1745721

Figure Lengend Snippet: (A) Comparison of apoptosis-related biological processes in THP-1 cells stimulated with F. alocis EVs or purified FtxA. Gene set enrichment analysis of RNA-seq data shows that EVs from the ftxA - strain 148B-17U (blue) upregulated multiple apoptosis-related pathways, including extrinsic and intrinsic apoptotic signaling and programmed cell death. In contrast, EVs from the ftx A + strain (orange), as well as purified FtxA (green) predominantly downregulated these pathways, indicating a suppressive effect on host cell apoptosis. (B) KEGG pathway enrichment analysis of apoptosis and necroptosis assessing EVs from ATCC 35896 ( ftxA + ) and 148B-17U ( ftxA − ), and recombinant FtxA, respectively. Negative enrichment scores (NES) are indicated in both panels.

Article Snippet: To further validate the immunosuppressive effects of FtxA, a multiplex cytokine array assay was conducted on THP-1 cell culture supernatants after exposure to purified FtxA, and EVs obtained from ATCC 35896 ( ftxA + ), and 148B-17U ( ftxA − ).

Techniques: Comparison, Purification, RNA Sequencing, Recombinant

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Filifactor alocis FtxA blocks inflammation and apoptosis pathways in monocytic cells

doi: 10.3389/fcimb.2026.1745721

Figure Lengend Snippet: Cytokine array analysis of THP-1 cell supernatants (two biological replicates) after stimulation with EVs from the F. alocis strains ATCC 35896 ( ftxA + ), 148B-17U, ( ftxA - ), and FtxA recombinant protein, respectively. Expression of pro- and anti-inflammatory cytokines (TNF-α, IL-8, IL-10, MIP-1α, MIP-1β, and MCP-1) is shown. A significant reduction in the secretion of TNF-α, IL-8, IL-10, MIP-1α, MIP-1β, and MCP-1, in cells treated with ATCC 35896 EVs and purified FtxA, respectively, compared with 148B-17U EVs (p < 0.05) was observed. Control represents untreated THP-1 cells. Data are presented as means ± standard deviation from three independent experiments.

Article Snippet: To further validate the immunosuppressive effects of FtxA, a multiplex cytokine array assay was conducted on THP-1 cell culture supernatants after exposure to purified FtxA, and EVs obtained from ATCC 35896 ( ftxA + ), and 148B-17U ( ftxA − ).

Techniques: Recombinant, Expressing, Purification, Control, Standard Deviation