Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Different Concentrations of Lactobacillus acidophilus Cell Free Filtrate Have Differing Anti-Biofilm and Immunomodulatory Effects

doi: 10.3389/fcimb.2021.737392

Figure Lengend Snippet: L. acidophilus cell free filtrate (CFF) has concentration-dependent effects on TNFα secretion in THP-1 monocytes and monocyte derived macrophages. THP-1 monocytes or monocyte derived macrophages were incubated with L. acidophilus 10 8 or 10 6 CFF alone or with cells 1 h after LPS stimulation. Supernatants were obtained 20 h after incubation and analyzed for levels of TNFα using ELISA. (A) L. acidophilus 10 8 CFF increased TNFα secretion in LPS-stimulated THP-1 monocytes, but 10 6 CFF had no effect. (B) In THP-1 monocyte derived macrophages, L. acidophilus 10 8 CFF alone promoted TNFα production while 10 6 CFF did not significantly alter TNFα production. (C) When THP-1 monocyte derived macrophages were stimulated with 50 ng/mL LPS 1 h prior to L. acidophilus CFF treatment, 10 6 CFF significantly attenuated TNFα production. Data are mean ± s.e.m. % of LPS Control data are mean ± s.e.m. percent change from LPS control adjusted to 100%. *p < 0.05, **p < 0.01 compared to controls; THP-1 monocytes n = 5 independent experiments; THP-1 monocyte derived macrophages n = 5 independent experiments; THP-1 monocyte derived macrophages % of LPS control n = 4 independent experiments.

Article Snippet: All bacterial strains as well as the THP-1 TIB-202 and THP-1 NF-κB-LUC2 cell lines were obtained from the American Type Culture Collection (Manassas, VA, USA) .

Techniques: Concentration Assay, Derivative Assay, Incubation, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Different Concentrations of Lactobacillus acidophilus Cell Free Filtrate Have Differing Anti-Biofilm and Immunomodulatory Effects

doi: 10.3389/fcimb.2021.737392

Figure Lengend Snippet: L. acidophilus cell free filtrate (CFF) affects NF-κB activity. Using a luciferase NF-κB reporter THP-1 cell line, monocytes and monocyte derived macrophages were incubated with L. acidophilus CFF alone or added 1 h after LPS stimulation, for 20 h. Luciferase luminescence was measured to quantify NF-κB activation. (A) In monocytes, 10 8 CFF alone upregulated NF-κB activation but 10 6 CFF had no effect. (B) In LPS stimulated monocytes, only 10 6 CFF reduced NF-κB activity. (C) L. acidophilus 10 8 CFF increased NF-κB activity in monocyte derived macrophages. (D) Similar to monocytes, only L. acidophilus 10 6 CFF significantly reduced NF-κB activity in monocyte derived macrophages. Data are relative light units (RLUs) or mean ± s.e.m. percent change from LPS control adjusted to 100%. **p < 0.01, ***p < 0.001 compared to appropriate control; monocytes n = 7 independent experiments; macrophages n = 8 independent experiments.

Article Snippet: All bacterial strains as well as the THP-1 TIB-202 and THP-1 NF-κB-LUC2 cell lines were obtained from the American Type Culture Collection (Manassas, VA, USA) .

Techniques: Activity Assay, Luciferase, Derivative Assay, Incubation, Activation Assay

Journal: FEBS letters

Article Title: Regulation of Interleukin-1 beta secretion from Macrophages via modulation of Potassium ion (K + ) channel activity

doi: 10.1002/1873-3468.13395

Figure Lengend Snippet: THP1 macrophages were differentiated with 100 nM PMA (Control), then loaded with AcLDL (50 µg/ml) and either used immediately after loading (+AcLDL) or 48 h after initiation of cholesterol efflux with medium containing 10% FBS as extracellular cholesterol acceptor (+AcLDL+Efflux). Panel A: Total cholesterol content of the cells after indicated treatments (Mean ± SD, n=3). Panel B: Current traces upon stepping for 500 ms from −100 mV to +60 mV in 10 mV increments from a holding potential of −60 mV under physiological K+ gradient (140 mM in the pipette and 5 mM in the bath). Panel C: Current-voltage (I-V) relationships (Mean ± SD, n=5 cells for each condition) from −100 mV to +60mV for indicated groups. Panel D: AUC for the positive currents for individual I-V curve included in Panel A was calculated and data are presented as Mean ± SD for the indicated groups. Panel E: Chord conductance was calculated as described under “Methods” and data are presented as Mean ± SD. One-way ANOVA and Tukey’s multiple comparison tests were performed and dissimilar letters indicate P<0.05.

Article Snippet: Human THP1 macrophages: THP1 monocytes obtained from ATCC were grown and differentiated using PMA as described ( 19 ).

Techniques: Transferring

Journal: FEBS letters

Article Title: Regulation of Interleukin-1 beta secretion from Macrophages via modulation of Potassium ion (K + ) channel activity

doi: 10.1002/1873-3468.13395

Figure Lengend Snippet: THP1 macrophages without any cholesterol loading (No AcLDL), with AcLDL loading (+AcLDL) or with stimulated efflux following AcLDL loading (+AcLDL+Efflux) were used to measure intracellular K+ levels and IL-1β secretion following treatment with LPS and Nigericin; untreated cells were used as No treatment control. Panel A: Mean Fluorescent Intensity of PBFI in 7AAD negative live cells was determined as a measure of intracellular K+ concentration and the data (Mean ± SD, n=6) are expressed as % No treatment control for the indicated groups. Panel B: LPS+Nigericin induced IL-1β secretion was determined and the data (Mean ± SD, n=6) are expressed as % No treatment Control. One-way ANOVA and Tukey’s multiple comparison tests were performed and dissimilar letters indicate P<0.05.

Article Snippet: Human THP1 macrophages: THP1 monocytes obtained from ATCC were grown and differentiated using PMA as described ( 19 ).

Techniques: Concentration Assay

Journal: FEBS letters

Article Title: Regulation of Interleukin-1 beta secretion from Macrophages via modulation of Potassium ion (K + ) channel activity

doi: 10.1002/1873-3468.13395

Figure Lengend Snippet: Panel A: Target channels for the inhibitors used. MPMs from C57BL/6 mice (Panel B) and THP1 macrophages (Panel C) were treated with LPS (50 ng/ml) overnight and then pre-treated with indicated K+ channel inhibitors for 1h. Nigericin (20 µM for 20 min) induced IL-1β secretion was determined. Data (Mean ± SD, n=6) are presented as %LPS + Nigericin. One-way ANOVA and Tukey’s multiple comparison tests were performed and dissimilar letters indicate P<0.05.

Article Snippet: Human THP1 macrophages: THP1 monocytes obtained from ATCC were grown and differentiated using PMA as described ( 19 ).

Techniques:

Journal: FEBS letters

Article Title: Regulation of Interleukin-1 beta secretion from Macrophages via modulation of Potassium ion (K + ) channel activity

doi: 10.1002/1873-3468.13395

Figure Lengend Snippet: MPMs from C57BL/6 mice (Panel A and B) and THP1 macrophages (Panel C and D) were treated with LPS (50 ng/ml) overnight and then pre-treated with indicated concentrations of Nateglinide or Repaglinide for 1h. Nigericin (20 µM for 20 min) induced IL-1β secretion was determined. Data (Mean ± SD, n=3) are presented as %LPS + Nigericin. One-way ANOVA and Tukey’s multiple comparison tests were performed and dissimilar letters indicate P<0.05.

Article Snippet: Human THP1 macrophages: THP1 monocytes obtained from ATCC were grown and differentiated using PMA as described ( 19 ).

Techniques:

Journal: FEBS letters

Article Title: Regulation of Interleukin-1 beta secretion from Macrophages via modulation of Potassium ion (K + ) channel activity

doi: 10.1002/1873-3468.13395

Figure Lengend Snippet: MPMs (Panel A) or THP1 macrophages (Panel B) were treated with LPS (50 ng/ml) overnight and then pre-treated with indicated K+ channel inhibitors for 1h. Nigericin (N, 20 µM for 20 min) or ATP (5 mM for 30 min) induced IL-1β secretion was determined. Data (Mean ± SD, n=6) are presented as %LPS + Nigericin or ATP. *P<0.05.

Article Snippet: Human THP1 macrophages: THP1 monocytes obtained from ATCC were grown and differentiated using PMA as described ( 19 ).

Techniques: