Journal: Cancer Immunology, Immunotherapy : CII

Article Title: NLRP3 in tumor-associated macrophages predicts a poor prognosis and promotes tumor growth in head and neck squamous cell carcinoma

doi: 10.1007/s00262-022-03357-4

Figure Lengend Snippet: Induction of HNSCC in Nlrp3-deficient mice by 4NQO. a Haematoxylin and eosin (H&E) staining showed the pathological changes in mouse tongues after 4NQO exposure. The dotted line indicates the epithelial basement membrane. Bar, 200 μm. b Schematic of the 4NQO tumorigenesis protocol. c Representative pictures of oral lesions in control and Nlrp3−/− mice at 24 weeks. d Representative H&E histological sections of 4NQO induced HNSCC. Bar, 50 μm. e Tumor incidence in the Nlrp3−/− (n = 10) and control (n = 10) groups. ns, no significant difference. f The proportion of oral lesion types in the two groups at week 24. g Representative image of tongue SCC in the two groups of mice. The red dotted circle denotes the lesion area. h Quantitative analysis of tumor size and i number in the Nlrp3−/− (n = 7) and control groups (n = 8) at the experimental endpoint. *, P < 0.05. j The body weights of 4NQO-treated control and Nlrp3−/− mice from the beginning to the end of this experiment. *, P < 0.05; ***, P < 0.001. Error bar, SD

Article Snippet: Flow Cytometry : PE anti-human/mouse NLRP3 (IC7578P) was obtained from R&D Systems.

Techniques: Staining, Membrane, Control

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: NLRP3 in tumor-associated macrophages predicts a poor prognosis and promotes tumor growth in head and neck squamous cell carcinoma

doi: 10.1007/s00262-022-03357-4

Figure Lengend Snippet: NLRP3 is overexpressed in the stromal cells of human HNSCC. a Representative immunofluorescence images of NLRP3 (red) and CK-14 (green) in human HNSCC tissue. Bar, 50 μm. b Flow cytometry to detect NLRP3 expression (mean ± SD) in HNSCC TILs, and cells were gated by CD14+. c, d Representative IHC images and corresponding quantification data of NLRP3 expression in oral mucosa (n = 20), dysplasia (n = 61) and HNSCC (n = 157) (***, P < 0.001). Bar, 50 μm. e Representative IHC staining of NLRP3 in HPV-negative (n = 142) and HPV-positive HNSCC tissue (n = 15) and f quantification analysis (**, P < 0.01). Bar, 50 μm. G Kaplan–Meier analysis using overall survival (OS) of NLRP3 expression in HNSCC patients. median cut-off, n = 148. Error bar, SD

Article Snippet: Flow Cytometry : PE anti-human/mouse NLRP3 (IC7578P) was obtained from R&D Systems.

Techniques: Immunofluorescence, Flow Cytometry, Expressing, Immunohistochemistry

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: NLRP3 in tumor-associated macrophages predicts a poor prognosis and promotes tumor growth in head and neck squamous cell carcinoma

doi: 10.1007/s00262-022-03357-4

Figure Lengend Snippet: Increased NLRP3 expression in TAMs predicts poor prognosis in HNSCC. a The ratio of M1 (CD64, CD68) and M2-like macrophage (MRC1, CD163) signatures in the low-expression (n = 259) and high-expression NLRP3 groups (n = 259) in the TCGA HNSCC database (***, P < 0.001). b Co-immunofluorescence staining of NLRP3 (red) and CD206 (green) in human HNSCC tissue. The white arrow shows co-expressed cell (yellow). T, tumor area; S, stromal area. Bar, 50 μm. c Flow cytometry to detect NLRP3 expression (mean ± SD) in CD206+ TAMs in human HNSCC PBMCs (n = 6) and TILs (n = 5). d Quantitative statistical analysis of NLRP3- and CD206-positive populations on gated cells (**, P < 0.01; ***, P < 0.001). e Correlation analysis of CD14+/CD206+/NLRP3+ and CD14+/CD206+ cell proportions in HNSCC patient PBMC and TIL, n = 11. f Representative IHC staining of NLRP3, CD206 and CD163 in human HNSCC using serial sections. Bar, 50 μm. g Correlation analysis between NLRP3, CD206 and CD163 protein expression in HNSCC tissue microarray using histoscore, n = 157. h Correlation analysis between NLRP3, MRC1 (CD206 encoding gene) and CD163 RNA expression in the TCGA HNSCC database. unit, expression (RSEM, Log2(Val + 1)), n = 520. i Hierarchical clustering plot of NLRP3, CD11b, CD206 and CD163 in HNSCC based on histoscore. n = 74. j Survival curves of high NLRP3 and CD206 expression vs. low NLRP3 and CD206 expression patients; high NLRP3 and CD163 expression vs. low NLRP3 and CD163 expression patients; low NLRP3 on high CD206 expression vs. high NLRP3 on CD206 high expression patients; and low NLRP3 on high CD163 expression vs. high NLRP3 on CD163 high expression patients. Error bar, SD

Article Snippet: Flow Cytometry : PE anti-human/mouse NLRP3 (IC7578P) was obtained from R&D Systems.

Techniques: Expressing, Immunofluorescence, Staining, Flow Cytometry, Immunohistochemistry, Microarray, RNA Expression

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: NLRP3 in tumor-associated macrophages predicts a poor prognosis and promotes tumor growth in head and neck squamous cell carcinoma

doi: 10.1007/s00262-022-03357-4

Figure Lengend Snippet: NLRP3 overexpression is positively correlated with TAMs in mouse HNSCC. a Representative IHC images of NLRP3, CD206 and F4/80 in Tgfbr1/Pten 2cKO and 4NQO-induced mouse HNSCC tumor tissues. Bar, 50 μm. b Correlation analysis between NLRP3, CD206 and F4/80 expression histoscores in mouse HNSCC tissues. n = 27. c Co-immunofluorescence staining of CD206 (green) and NLRP3 (red) in mouse HNSCC tissue. Bar, 50 μm. d, e Representative flow cytometry plots and quantification analysis showed the population change of NLRP3+ TAMs in the blood of normal wild-type and tumor-bearing individuals in two mouse models (n = 6 in 4NQO; n = 4 in 2cKO) (***, P < 0.001). Error bar, SD

Article Snippet: Flow Cytometry : PE anti-human/mouse NLRP3 (IC7578P) was obtained from R&D Systems.

Techniques: Over Expression, Expressing, Immunofluorescence, Staining, Flow Cytometry

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: NLRP3 in tumor-associated macrophages predicts a poor prognosis and promotes tumor growth in head and neck squamous cell carcinoma

doi: 10.1007/s00262-022-03357-4

Figure Lengend Snippet: Knockdown of NLRP3 in THP-1 cells inhibits M2-like macrophage polarization. a THP-1 cells were treated with CAL27 cultured medium (CM) or IL-4 and IL-13 after treatment with PMA; NLRP3 and Caspase-1 expression were analyzed by western blot, and b IL-1β amounts in the supernatant were detected by ELISA (n = 3 per group; ***, P < 0.001). c Western blotting analysis of NLRP3 expression in NLRP3-knockout or control THP-1 cells. d, e Flow cytometry plots and quantification analysis of CD206 in NLRP3-deficient and control THP-1 macrophages after CAL27 CM culture (n = 3 per group; ***, P < 0.001). f ELISA results of IL-1β amounts in control and siNLRP3 THP-1 macrophage cultured medium (n = 3 per group; ***, P < 0.001). g, h NLRP3-deficient THP-1 macrophages were exposed to IL-1β (20 ng/ml) for 48 h after CAL27 CM treatment, and CD206 was analyzed by flow cytometry (n = 3 per group; ***, P < 0.001). i Growth curves of CAL27 cells in different groups as measured by a CCK-8 assay (n = 3 per group; ***, P < 0.001). Error bar, SD

Article Snippet: Flow Cytometry : PE anti-human/mouse NLRP3 (IC7578P) was obtained from R&D Systems.

Techniques: Knockdown, Cell Culture, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Knock-Out, Control, Flow Cytometry, CCK-8 Assay

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: NLRP3 in tumor-associated macrophages predicts a poor prognosis and promotes tumor growth in head and neck squamous cell carcinoma

doi: 10.1007/s00262-022-03357-4

Figure Lengend Snippet: NLRP3 deficiency reduced the proportion of TAMs and MDSCs. a Representative IHC images and quantification of IL-1β in tumor tissues from the two groups of mice. ***, P < 0.001, Bar, 50 μm. b ELISA results of IL-1β amounts in the blood of normal WT (n = 5), 4NQO-treated control (n = 8) and Nlrp3−/− (n = 7) tumor burden mice. **, P < 0.01; ***, P < 0.001; ns, no significant difference. c The Luminex liquid suspension chip analysis result. *, P < 0.05; **, P < 0.01, three mice in each group. d Flow cytometry to detect the percentage of CD11b+/F4/80+/CD206+ TAMs in the spleen of the two groups and e quantitative statistical analysis. *, P < 0.05. f Representative IHC images and quantification of CD206 in control and Nlrp3−/− mouse HNSCC tissues. **, P < 0.01, Bar, 50 μm. g Flow cytometry to detect the percentage of M-MDSCs and PMN-MDSCs in the spleen of the two groups and h quantitative statistical analysis. *, P < 0.05. i Representative IHC images and quantification of Gr-1 in control and Nlrp3−/− mouse HNSCC tissues. **, P < 0.01, Bar, 50 μm

Article Snippet: Flow Cytometry : PE anti-human/mouse NLRP3 (IC7578P) was obtained from R&D Systems.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Luminex, Suspension, Flow Cytometry

Journal: Frontiers in Immunology

Article Title: Distinct SARS-CoV-2 specific NLRP3 and IL-1β responses in T cells of aging patients during acute COVID-19 infection

doi: 10.3389/fimmu.2023.1231087

Figure Lengend Snippet: SPIKE-specific T cell-IL-1β expression is reduced in COVID+ patients aged ≥ 61. PBMCs from age ≤ 60 (n=20) and aged ≥ 61 (n=16) COVID+ patients were stimulated with SPIKE peptides in vitro for 12 h to assess COVID-specific responses in T cells. Representative contour plots showing % of PD-1 and IFN-γ expressing (A, B) , T-bet (D) , NLRP3 (E) , of CD4+ (above) and CD8+ (below) cells. Statistics showing % IFN-γ/ %PD-1 ratios (C) and NLRP3 expression (F) , in CD4+ (top) and CD8+ cells (bottom) in PBMCs from subjects aged ≤ 60 and ≥ 61 in vitro. Median values ± SEM are plotted. (Mann-Whitney U test; Two-tailed). ***<0.0005, ****<0.00005.

Article Snippet: NLRP3-PE (bs-10021R-PE) is from Bioss (MN, US).

Techniques: Expressing, In Vitro, MANN-WHITNEY, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Distinct SARS-CoV-2 specific NLRP3 and IL-1β responses in T cells of aging patients during acute COVID-19 infection

doi: 10.3389/fimmu.2023.1231087

Figure Lengend Snippet: IL-1β expression positively correlates with IFN-γ and NLRP3 expression in SPIKE-activated T cells in COVID+ patients. Spearman Correlation (r) analysis showing positive correlation between IL-1β and IFN-γ in CD4 + T cells (A) and CD8 + T cells (B) , and NLRP3 and IFN-γ in CD4 + T cells (C) , and CD8 + T cells (D) . Negative correlation between cytokines and age in CD4 + T cells (E, G) and CD8 + T cells (F, H) . *P<0.05, **<0.005, ***<0.0005, ****<0.00005.

Article Snippet: NLRP3-PE (bs-10021R-PE) is from Bioss (MN, US).

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: Distinct SARS-CoV-2 specific NLRP3 and IL-1β responses in T cells of aging patients during acute COVID-19 infection

doi: 10.3389/fimmu.2023.1231087

Figure Lengend Snippet: Cytokine dysregulation is not observed in aged monocytes (aged ≥ 61) in SPIKE-activated PBMCs from COVID-infected individuals. PBMCs from COVID+ patients aged ≤ 60 and ≥ 61 were stimulated with SPIKE for 12 h as in Figure 2 . Representative contour plots (A) showing % of IL-1β (x-axis) and IFN-γ (y-axis) gating on CD14 + monocytes and statistics of % IL-1β + cells in COVID+ patients aged ≤ 60 and ≥ 61 (B) . Histogram plots showing NLRP3 expression (C) and statistical analysis of % of NLRP3 + cells in COVID+ patients (D) . Median values ± SEM are plotted. (Mann-Whitney U test; Two-tailed). NS, non significant.

Article Snippet: NLRP3-PE (bs-10021R-PE) is from Bioss (MN, US).

Techniques: Infection, Expressing, MANN-WHITNEY, Two Tailed Test