Journal: PNAS Nexus

Article Title: Soluble CD14 promotes Th17 expansion and differentiation through gamma-aminobutyric acid and expands infidel innate lymphoid cells

doi: 10.1093/pnasnexus/pgaf406

Figure Lengend Snippet: The supernatant of sCD14-treated PBMCs promotes Th17 phenotype. A) The schematic plot of the experimental design. B) Volcano plot of differentially expressed genes in isolated T cells stimulated with anti-CD3/CD28, with and without supernatant from sCD14-treated PBMCs. C) Heat map comparing differentially expressed genes associated with Th17 cells in isolated T cells stimulated with anti-CD3/CD28 plus supernatant from unstimulated autologous PBMCs, and treated with the supernatant from sCD14-treated PBMCs. D) Abundance (TPM) of various genes associated with Th17 cells in isolated T cells, with or without treatment with the supernatant from sCD14-treated PBMCs. E) Bar plot showing the up- and down-regulated transcriptional factors in isolated T cells after treatment with the supernatant from sCD14-treated PBMCs. F) Fold change in IL-17A gene expression in isolated T cells after treatment with the supernatant from sCD14-treated PBMCs. G) IL-17A levels in culture supernatants of isolated T cells treated with the supernatant from sCD14-treated PBMCs, as quantified by ELISA. H) Isolated T cells (1 × 10 5 ) were stimulated in the presence of the supernatant from sCD14-treated PBMCs, and IL-17A-secreting cells were visualized and enumerated by ELISpot. I) Box plot showing the expression of CTLA-4 gene in T cells stimulated with anti-CD3/CD28, in the presence and absence of the supernatant from sCD14-treated PBMCs. P values were calculated using two-tailed Mann–Whitney U test (D, F, I) and Wilcoxon matched-pairs signed-rank tests (G, H); * P < 0.05, **<0.01, ****<0.0001. Each dot represents sample from a study subject.

Article Snippet: The concentrations of IL-17A and sCD14 were measured by DuoSet ELISA kit (R&D systems, Cat# DY317 and Cat# DY383-05, respectively) following the manufacturer's protocols.

Techniques: Isolation, Gene Expression, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot, Expressing, Two Tailed Test, MANN-WHITNEY

Journal: PNAS Nexus

Article Title: Soluble CD14 promotes Th17 expansion and differentiation through gamma-aminobutyric acid and expands infidel innate lymphoid cells

doi: 10.1093/pnasnexus/pgaf406

Figure Lengend Snippet: Increased GABA in the supernatant of sCD14-treated monocytes promotes IL-17A secretion. A) Volcano plot illustrating the magnitude and significance of differences in cytokine/chemokine concentrations in the supernatant of isolated monocytes from HCs ( n = 10) with and without overnight treatment with sCD14. B) Normalized and calculated concentrations of shown cytokines in the supernatant of isolated monocytes from HCs with and without overnight treatment with sCD14. C) Metabolic pathway enrichment analysis plot showing significantly altered pathways in the supernatant of monocytes isolated from 12 HCs (six males, six females) after treatment with sCD14, with box plots showing the metabolites altered in each pathway. D) Metabolic pathway enrichment analysis plot showing significantly altered pathways in the supernatant of monocytes isolated from male HCs ( n = 6) and E) female HCs ( n = 6) after treatment with sCD14, with box plots showing the metabolites altered in each pathway. F) Culture supernatant concentrations of IL-17A in isolated T cells treated with the supernatant from sCD14-treated PBMCs, GABA, hydroxyproline, arginine, and Th17-polarizing conditions, as quantified by ELISA. G) Fold change in IL17A gene expression in isolated T cells after treatment with the supernatant from sCD14-treated PBMCs and GABA. P values were calculated using two-tailed Mann–Whitney U test (C, D, E), Kruskal–Wallis analysis with Dunn's multiple comparisons test (G); * P < 0.05, **<0.01, ***<0.001. Each dot represents sample from a human study subject.

Article Snippet: The concentrations of IL-17A and sCD14 were measured by DuoSet ELISA kit (R&D systems, Cat# DY317 and Cat# DY383-05, respectively) following the manufacturer's protocols.

Techniques: Isolation, Enzyme-linked Immunosorbent Assay, Gene Expression, Two Tailed Test, MANN-WHITNEY