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BNP treatment increased moLCs migration toward CCL21 and reduced <t>chemokine</t> production in activated moLCs. Monocytes were cultured in the presence of GMCSF, TNF-α, and TGF-β for 5 days supplemented with IL-4 for the first 48 hrs to differentiate moLCs, and treated with 10 nM BNP throughout the entire differentiation process. On day 4, moLCs were activated with CL075 and poly(I:C) and a combination of both for 24 hrs. (A) In Transwell migration assay, 1*10 6 cells were applied in the upper well of the Transwell plate, and in the bottom of the plate, it contains RPMI supplemented with CCL19 and CCL21. Migration of the cells was measured with flow cytometry. Data are presented as individual values with mean ± SEM. Statistical significance was determined by Two-way ANOVA followed by Tukey’s multiple comparison test. Normality of residuals was confirmed by QQ plot analysis. n=8 biological replicates per group (B) A chemokine array was performed using supernatants from BNP treated and TLR activated cells. Symbols with different colors represent individual donors. Data is presented as Mean ± SEM. One-way ANOVA with Tukey’s multiple comparison test was used for statistical analysis. n=2 biological replicates per group *P<0.05, **P<0.01, *** P<0.001, **** P< 0.0001. BNP, B-type natriuretic peptide; CL075; TLR7/8 agonist, thiazoquinoline compound; CTRL, Control; moLC, monocyte-derived Langerhans cells; Poly(I:C), TLR3 agonist, polyinosinic:polycytidylic acid.
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BNP treatment increased moLCs migration toward CCL21 and reduced <t>chemokine</t> production in activated moLCs. Monocytes were cultured in the presence of GMCSF, TNF-α, and TGF-β for 5 days supplemented with IL-4 for the first 48 hrs to differentiate moLCs, and treated with 10 nM BNP throughout the entire differentiation process. On day 4, moLCs were activated with CL075 and poly(I:C) and a combination of both for 24 hrs. (A) In Transwell migration assay, 1*10 6 cells were applied in the upper well of the Transwell plate, and in the bottom of the plate, it contains RPMI supplemented with CCL19 and CCL21. Migration of the cells was measured with flow cytometry. Data are presented as individual values with mean ± SEM. Statistical significance was determined by Two-way ANOVA followed by Tukey’s multiple comparison test. Normality of residuals was confirmed by QQ plot analysis. n=8 biological replicates per group (B) A chemokine array was performed using supernatants from BNP treated and TLR activated cells. Symbols with different colors represent individual donors. Data is presented as Mean ± SEM. One-way ANOVA with Tukey’s multiple comparison test was used for statistical analysis. n=2 biological replicates per group *P<0.05, **P<0.01, *** P<0.001, **** P< 0.0001. BNP, B-type natriuretic peptide; CL075; TLR7/8 agonist, thiazoquinoline compound; CTRL, Control; moLC, monocyte-derived Langerhans cells; Poly(I:C), TLR3 agonist, polyinosinic:polycytidylic acid.
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BNP treatment increased moLCs migration toward CCL21 and reduced <t>chemokine</t> production in activated moLCs. Monocytes were cultured in the presence of GMCSF, TNF-α, and TGF-β for 5 days supplemented with IL-4 for the first 48 hrs to differentiate moLCs, and treated with 10 nM BNP throughout the entire differentiation process. On day 4, moLCs were activated with CL075 and poly(I:C) and a combination of both for 24 hrs. (A) In Transwell migration assay, 1*10 6 cells were applied in the upper well of the Transwell plate, and in the bottom of the plate, it contains RPMI supplemented with CCL19 and CCL21. Migration of the cells was measured with flow cytometry. Data are presented as individual values with mean ± SEM. Statistical significance was determined by Two-way ANOVA followed by Tukey’s multiple comparison test. Normality of residuals was confirmed by QQ plot analysis. n=8 biological replicates per group (B) A chemokine array was performed using supernatants from BNP treated and TLR activated cells. Symbols with different colors represent individual donors. Data is presented as Mean ± SEM. One-way ANOVA with Tukey’s multiple comparison test was used for statistical analysis. n=2 biological replicates per group *P<0.05, **P<0.01, *** P<0.001, **** P< 0.0001. BNP, B-type natriuretic peptide; CL075; TLR7/8 agonist, thiazoquinoline compound; CTRL, Control; moLC, monocyte-derived Langerhans cells; Poly(I:C), TLR3 agonist, polyinosinic:polycytidylic acid.
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BNP treatment increased moLCs migration toward CCL21 and reduced chemokine production in activated moLCs. Monocytes were cultured in the presence of GMCSF, TNF-α, and TGF-β for 5 days supplemented with IL-4 for the first 48 hrs to differentiate moLCs, and treated with 10 nM BNP throughout the entire differentiation process. On day 4, moLCs were activated with CL075 and poly(I:C) and a combination of both for 24 hrs. (A) In Transwell migration assay, 1*10 6 cells were applied in the upper well of the Transwell plate, and in the bottom of the plate, it contains RPMI supplemented with CCL19 and CCL21. Migration of the cells was measured with flow cytometry. Data are presented as individual values with mean ± SEM. Statistical significance was determined by Two-way ANOVA followed by Tukey’s multiple comparison test. Normality of residuals was confirmed by QQ plot analysis. n=8 biological replicates per group (B) A chemokine array was performed using supernatants from BNP treated and TLR activated cells. Symbols with different colors represent individual donors. Data is presented as Mean ± SEM. One-way ANOVA with Tukey’s multiple comparison test was used for statistical analysis. n=2 biological replicates per group *P<0.05, **P<0.01, *** P<0.001, **** P< 0.0001. BNP, B-type natriuretic peptide; CL075; TLR7/8 agonist, thiazoquinoline compound; CTRL, Control; moLC, monocyte-derived Langerhans cells; Poly(I:C), TLR3 agonist, polyinosinic:polycytidylic acid.

Journal: Frontiers in Immunology

Article Title: B-type natriuretic peptide attenuates TLR-induced cytokine and chemokine secretion in monocyte-derived Langerhans cells

doi: 10.3389/fimmu.2026.1791284

Figure Lengend Snippet: BNP treatment increased moLCs migration toward CCL21 and reduced chemokine production in activated moLCs. Monocytes were cultured in the presence of GMCSF, TNF-α, and TGF-β for 5 days supplemented with IL-4 for the first 48 hrs to differentiate moLCs, and treated with 10 nM BNP throughout the entire differentiation process. On day 4, moLCs were activated with CL075 and poly(I:C) and a combination of both for 24 hrs. (A) In Transwell migration assay, 1*10 6 cells were applied in the upper well of the Transwell plate, and in the bottom of the plate, it contains RPMI supplemented with CCL19 and CCL21. Migration of the cells was measured with flow cytometry. Data are presented as individual values with mean ± SEM. Statistical significance was determined by Two-way ANOVA followed by Tukey’s multiple comparison test. Normality of residuals was confirmed by QQ plot analysis. n=8 biological replicates per group (B) A chemokine array was performed using supernatants from BNP treated and TLR activated cells. Symbols with different colors represent individual donors. Data is presented as Mean ± SEM. One-way ANOVA with Tukey’s multiple comparison test was used for statistical analysis. n=2 biological replicates per group *P<0.05, **P<0.01, *** P<0.001, **** P< 0.0001. BNP, B-type natriuretic peptide; CL075; TLR7/8 agonist, thiazoquinoline compound; CTRL, Control; moLC, monocyte-derived Langerhans cells; Poly(I:C), TLR3 agonist, polyinosinic:polycytidylic acid.

Article Snippet: Proteome Profiler Human Chemokine Array Kit (R&D Systems) was performed according to the manufacturer’s instructions.

Techniques: Migration, Cell Culture, Transwell Migration Assay, Flow Cytometry, Comparison, Control, Derivative Assay