cd4 t cell isolation kit  (Miltenyi Biotec)

Journal: Cell Death Discovery

Article Title: Epithelial pyroptosis-induced TREM1 + macrophages activate Th17 cells to accelerate oral mucosal inflammation

doi: 10.1038/s41420-025-02853-7

Figure Lengend Snippet: a Top left, UMAP plot of 9542 cell colors by myeloid cell types. Gray circle: Mφ. Bottom left, UMAP plot displayed the differential abundance (DA) myeloid cells of the inflammatory development of oral mucosal epithelium. Red: DA myeloid cells in REOLP. Blue: DA myeloid cells in NREOLP. Gray circle: DA Mφ in REOLP. Top right, UMAP plot of 6,680 cell colors by Mφ types. Gray circle: DA Mφ in REOLP. Bottom right, violin plot of the expression of IL1B in different Mφ subtypes. b Scatterplot and network plot showing the active transcription factors of TREM1 + Mφ and the PPI of active transcription factors and IL-1β. c Ridge plots showed the differential expression of signaling pathways in TREM1 + Mφ and TREM1 - Mφ. d Circle plot of the intercellular communication (IC) from pyroptotic epithelial cells (top) and TREM1 + Mφ (bottom) to other cells in the inflammatory development of oral mucosal epithelium. Line width, the IC strength. Red arrow, the IC strength in REOLP was higher than in NREOLP. Blue arrow, the IC strength in NREOLP was higher than in REOLP. e Top left, UMAP plot of 75,336 cells colors by NK/T cell types. Gray circle: Th17 cells. Bottom left, UMAP plot displayed the DA NK/T cells of the inflammatory development of oral mucosal epithelium. Red: DA NK/T cells in REOLP. Blue: DA NK/T cells in NREOLP. Gray circle: DA Th17 cells in REOLP. Right, UMAP plot of 10,954 cell colors by Th17 cell types. Gray circle: DA Th17 cells in REOLP. f Left, dot plot of the IC from TREM1 + Mφ to naïve CD4 + T cells in IL1 signaling pathway in the inflammatory development of oral mucosal epithelium. Dot size, p-value; color scale, IC probability. Right, the developmental trajectory of T cell subtypes. g Bar plot exhibiting the upregulated signaling pathways of pathogenic Th17 cells in the inflammatory development of oral mucosal epithelium.

Article Snippet: Murine naïve CD4 + cells were obtained from mouse spleen and lymph nodes through grinding and passed through a 70 μM filter membrane (Biosharp, China, #23116232), lysed using red cell lysis buffer (Biosharp, China, #BL503B), and sorted using magnetic beads of CD62L + CD4 + T cells (Miltenyi Biotec, German, #130-106-643).

Techniques: Expressing, Quantitative Proteomics, Protein-Protein interactions

Journal: Cell Death Discovery

Article Title: Epithelial pyroptosis-induced TREM1 + macrophages activate Th17 cells to accelerate oral mucosal inflammation

doi: 10.1038/s41420-025-02853-7

Figure Lengend Snippet: a Flow chart of cytological experiments. b Flow cytometry showing the TREM1 expression of the TREM1 + Mφ and TREM1 NC Mφ. c Elisa assay depicting the concentration of IL-1β in TREM1 + Mφ and TREM1 NC Mφ with/without the stimulation of recombinant human HMGB1 (rhHMGB1), respectively. Note: ** p < 0.01, NS not statistically significant. Flow cytometry ( d ) and statistical analysis ( e ) of the c e ll proportion of CD4 + IL-17A + cells, CD4 + IL-17F + cells, and CD4 + IL-22 + cells after murine naïve CD4 + T cells co - cultured with the supernatants of TREM1 + Mφ, IL-6 and IL-23, with/without neutralizing anti-IL-1β (α-IL-1β). Note: * p < 0.05, ** p < 0.01, NS not statistically significant. f Representative images of mIHC in NREOLP (top) and REOLP (bottom). Scale bars: 100 µM (top) and 50 µM (bottom). Yellow arrows pointed to LPS + GSDMD + E-cad + cells. Red arrows pointed to TREM1 + CD68 + cells. Green arrows pointed to IL-17A + CD3 + cells.

Article Snippet: Murine naïve CD4 + cells were obtained from mouse spleen and lymph nodes through grinding and passed through a 70 μM filter membrane (Biosharp, China, #23116232), lysed using red cell lysis buffer (Biosharp, China, #BL503B), and sorted using magnetic beads of CD62L + CD4 + T cells (Miltenyi Biotec, German, #130-106-643).

Techniques: Flow Cytometry, Expressing, Enzyme-linked Immunosorbent Assay, Concentration Assay, Recombinant, Cell Culture