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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) , <t>NLRP3</t> (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.

Nlrp3 Pe, supplied by Bioss, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Distinct SARS-CoV-2 specific NLRP3 and IL-1β responses in T cells of aging patients during acute COVID-19 infection"

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

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2023.1231087

Figure Legend 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.

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

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.

Figure Legend 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.

Techniques Used: Expressing

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 <xref ref-type=

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. " title="... and ≥ 61 (B) . Histogram plots showing NLRP3 expression (C) and statistical analysis of % of ..." property="contentUrl" width="100%" height="100%"/>
Figure Legend 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.

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

Image Search Results

Influence of a black carrot extract (BCE) with high amounts of acylated anthocyanins and their related phenolic acids on ASC and NLRP3 protein expression in THP‐1 monocytes. THP‐1 monocytes were preincubated with the indicated concentrations of the BCE, C3G, or phenolic acids before cells were primed with LPS. A, C) ASC and B, D) NLRP3 protein expression were assessed as median fluorescence intensity (MFI) by intracellular flow cytometry. Significant differences to LPS primed cells were calculated using one‐way ANOVA with Dunnett's multiple comparisons test (** p < 0.01). ANOVA, analysis of variance; ASC, apoptosis‐associated speck‐like protein containing a caspase recruitment domain; cyanidin‐3‐glucoside (C3G); p ‐coumaric acid (CA); ferulic acid (FA); LPS, lipopolysaccharide; NLRP3, nucleotide‐binding oligomerization domain‐like receptor pyrin domain‐containing protein 3; sinapinic acid (SA).

Journal: Molecular Nutrition & Food Research

Article Title: Acylated Anthocyanins From Black Carrots and Their Related Phenolic Acids Diminish Priming and Activation of the NLRP3 Inflammasome in THP‐1 Monocytes

doi: 10.1002/mnfr.202400356

Figure Lengend Snippet: Influence of a black carrot extract (BCE) with high amounts of acylated anthocyanins and their related phenolic acids on ASC and NLRP3 protein expression in THP‐1 monocytes. THP‐1 monocytes were preincubated with the indicated concentrations of the BCE, C3G, or phenolic acids before cells were primed with LPS. A, C) ASC and B, D) NLRP3 protein expression were assessed as median fluorescence intensity (MFI) by intracellular flow cytometry. Significant differences to LPS primed cells were calculated using one‐way ANOVA with Dunnett's multiple comparisons test (** p < 0.01). ANOVA, analysis of variance; ASC, apoptosis‐associated speck‐like protein containing a caspase recruitment domain; cyanidin‐3‐glucoside (C3G); p ‐coumaric acid (CA); ferulic acid (FA); LPS, lipopolysaccharide; NLRP3, nucleotide‐binding oligomerization domain‐like receptor pyrin domain‐containing protein 3; sinapinic acid (SA).

Article Snippet: On the next day, cells were intracellularly stained with fluorescence‐labeled monoclonal antibodies against ASC (BioLegend) and NLRP3 (Miltenyi Biotec B.V. & Co. KG, Bergisch Gladbach, Germany) or the corresponding isotype controls.

Techniques: Expressing, Fluorescence, Flow Cytometry, Binding Assay

Influence of a black carrot extract (BCE) with high amounts of acylated anthocyanins and their related phenolic acids on ASC speck formation in THP‐1 monocytes. THP‐1 monocytes were preincubated with the indicated concentrations of the BCE, cyanidin‐3‐glucoside, or phenolic acids before the NLRP3 inflammasome was activated. Significant differences to A, C) LPS and nigericin stimulated cells or B, D) only nigericin treated cells were calculated using one‐way ANOVA with Dunnett's multiple comparisons test (* p < 0.05 and ** p < 0.01). ANOVA, analysis of variance; ASC, apoptosis‐associated speck‐like protein containing a caspase recruitment domain; C3G; CA; FA; LPS, lipopolysaccharide; NLRP3, nucleotide‐binding oligomerization domain‐like receptor pyrin domain‐containing protein 3; SA.

Journal: Molecular Nutrition & Food Research

Article Title: Acylated Anthocyanins From Black Carrots and Their Related Phenolic Acids Diminish Priming and Activation of the NLRP3 Inflammasome in THP‐1 Monocytes

doi: 10.1002/mnfr.202400356

Figure Lengend Snippet: Influence of a black carrot extract (BCE) with high amounts of acylated anthocyanins and their related phenolic acids on ASC speck formation in THP‐1 monocytes. THP‐1 monocytes were preincubated with the indicated concentrations of the BCE, cyanidin‐3‐glucoside, or phenolic acids before the NLRP3 inflammasome was activated. Significant differences to A, C) LPS and nigericin stimulated cells or B, D) only nigericin treated cells were calculated using one‐way ANOVA with Dunnett's multiple comparisons test (* p < 0.05 and ** p < 0.01). ANOVA, analysis of variance; ASC, apoptosis‐associated speck‐like protein containing a caspase recruitment domain; C3G; CA; FA; LPS, lipopolysaccharide; NLRP3, nucleotide‐binding oligomerization domain‐like receptor pyrin domain‐containing protein 3; SA.

Techniques: Binding Assay

Influence of a black carrot extract (BCE) with high amounts of acylated anthocyanins on caspase‐1 activity in THP‐1 monocytes. THP‐1 monocytes were preincubated with the indicated concentrations of the BCE or C3G before the NLRP3 inflammasome was activated. Caspase‐1 activity was measured by using a bioluminescent assay and luminescence was measured as relative light unit (RLU). Significant differences to (A) LPS and nigericin stimulated cells or (B) only nigericin treated cells were calculated using one‐way ANOVA with Dunnett's multiple comparisons test (** p < 0.01, *** p < 0.001, and **** p < 0.0001). ANOVA, analysis of variance; C3G, cyanidin‐3‐glucoside; LPS, lipopolysaccharide; NLRP3, nucleotide‐binding oligomerization domain‐like receptor pyrin domain‐containing protein 3.

Journal: Molecular Nutrition & Food Research

Article Title: Acylated Anthocyanins From Black Carrots and Their Related Phenolic Acids Diminish Priming and Activation of the NLRP3 Inflammasome in THP‐1 Monocytes

doi: 10.1002/mnfr.202400356

Figure Lengend Snippet: Influence of a black carrot extract (BCE) with high amounts of acylated anthocyanins on caspase‐1 activity in THP‐1 monocytes. THP‐1 monocytes were preincubated with the indicated concentrations of the BCE or C3G before the NLRP3 inflammasome was activated. Caspase‐1 activity was measured by using a bioluminescent assay and luminescence was measured as relative light unit (RLU). Significant differences to (A) LPS and nigericin stimulated cells or (B) only nigericin treated cells were calculated using one‐way ANOVA with Dunnett's multiple comparisons test (** p < 0.01, *** p < 0.001, and **** p < 0.0001). ANOVA, analysis of variance; C3G, cyanidin‐3‐glucoside; LPS, lipopolysaccharide; NLRP3, nucleotide‐binding oligomerization domain‐like receptor pyrin domain‐containing protein 3.

Techniques: Activity Assay, Binding Assay

Influence of a black carrot extract (BCE) with high amounts of acylated anthocyanins on proinflammatory cytokine release in THP‐1 monocytes. THP‐1 monocytes were preincubated with the indicated concentrations of BCE or C3G before the NLRP3 inflammasome was activated. Release of (A) IL‐1β and (B) IL‐18 into the cell culture supernatant was measured by ELISA. Significant differences to LPS and nigericin‐stimulated cells were calculated using one‐way ANOVA with Dunnett's multiple comparisons test (* p < 0.05 and ** p < 0.01). ANOVA, analysis of variance; C3G, cyanidin‐3‐glucoside; CA, p ‐coumaric acid; ELISA, enzyme‐linked immunosorbent assays; FA, ferulic acid; LPS, lipopolysaccharide; NLRP3, nucleotide‐binding oligomerization domain‐like receptor pyrin domain‐containing protein 3; SA, sinapinic acid.

Journal: Molecular Nutrition & Food Research

Article Title: Acylated Anthocyanins From Black Carrots and Their Related Phenolic Acids Diminish Priming and Activation of the NLRP3 Inflammasome in THP‐1 Monocytes

doi: 10.1002/mnfr.202400356

Figure Lengend Snippet: Influence of a black carrot extract (BCE) with high amounts of acylated anthocyanins on proinflammatory cytokine release in THP‐1 monocytes. THP‐1 monocytes were preincubated with the indicated concentrations of BCE or C3G before the NLRP3 inflammasome was activated. Release of (A) IL‐1β and (B) IL‐18 into the cell culture supernatant was measured by ELISA. Significant differences to LPS and nigericin‐stimulated cells were calculated using one‐way ANOVA with Dunnett's multiple comparisons test (* p < 0.05 and ** p < 0.01). ANOVA, analysis of variance; C3G, cyanidin‐3‐glucoside; CA, p ‐coumaric acid; ELISA, enzyme‐linked immunosorbent assays; FA, ferulic acid; LPS, lipopolysaccharide; NLRP3, nucleotide‐binding oligomerization domain‐like receptor pyrin domain‐containing protein 3; SA, sinapinic acid.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Binding Assay

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

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

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

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

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

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

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

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

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

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

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

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

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

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