Journal: Microorganisms

Article Title: Transcriptomic and Immunopathological Profiles of Inflammasomes in Different Clinical Forms of American Cutaneous Leishmaniasis

doi: 10.3390/microorganisms13050980

Figure Lengend Snippet: Boxplots showing normalized gene expression data from RNA-seq of lesions in patients with different clinical forms of American Cutaneous Leishmaniasis caused by L. (L.) amazonensis and L. (V.) braziliensis. ADCL: Anergic Diffuse Cutaneous Leishmaniasis, caused by L. (L.) amazonensis ; LCL (La): Localized Cutaneous Leishmaniasis, caused by L. (L.) amazonensis ; LCL (Lb): Localized Cutaneous Leishmaniasis, caused by L. (V.) braziliensis ; and MCL: Mucocutaneous Leishmaniasis caused by L. (V.) braziliensis. A: NLRP3; B: AIM2; C: NLRP12; D: NLRC4; E: CASP1; F: CASP5; G: GSDMD; H: IL-18; I: IL-1 β. * = p < 0.05; ** = p < 0.001 and *** = p < 0.0001.

Article Snippet: The sections were incubated with primary antibodies anti-NLRP3 (polyclonal, E-AB-70161, Elabscience, Houston, TX, USA, 1:600), anti-AIM-2 (polyclonal, E-AB-10974, Elabscience, 1:100), anti-NLRC4 (polyclonal, E-AB-67860, Elabscience, 1:100), anti-NLRP12 (polyclonal, E-AB-66298, Elabscince, 1:500), anti-caspase-1 (monoclonal, G6231-3D2, Sigma Aldrich, St. Louis, MO, USA, 1:500), anti-caspase-5 (monoclonal, MAA770Hu22, Cloud-clone Corp., Katy, TX, USA, 1:100), anti-GSDMD (monoclonal, E-AB-81473, Elabscience, 1:200), anti-IL-1β (polyclonal, ab2105, Abcam, Cambridge, MA, USA, 1:800), and anti-IL-18 (polyclonal, ab68435, Abcam, 1:2000).

Techniques: Gene Expression, RNA Sequencing

Journal: Microorganisms

Article Title: Transcriptomic and Immunopathological Profiles of Inflammasomes in Different Clinical Forms of American Cutaneous Leishmaniasis

doi: 10.3390/microorganisms13050980

Figure Lengend Snippet: Immunohistochemistry reactions on histological sections of biopsies from patients with different clinical forms of ACL using anti-NLRP3, anti-AIM-2, anti-NLRP12, anti-NLRC4, anti-caspase-1, anti-caspase-5, anti-GSDMD, anti-IL-1β, and anti-IL-18 antibodies. ADCL: Anergic Diffuse Cutaneous Leishmaniasis caused by L. (L.) amazonensis ; LCL (La): Localized Cutaneous Leishmaniasis caused by L. (L.) amazonensis ; LCL (Lb): Localized Cutaneous Leishmaniasis caused by L. (V.) braziliensis ; MCL: Mucocutaneous Leishmaniasis caused by L. (V.) braziliensis . Red arrows indicate immunostained cells.

Article Snippet: The sections were incubated with primary antibodies anti-NLRP3 (polyclonal, E-AB-70161, Elabscience, Houston, TX, USA, 1:600), anti-AIM-2 (polyclonal, E-AB-10974, Elabscience, 1:100), anti-NLRC4 (polyclonal, E-AB-67860, Elabscience, 1:100), anti-NLRP12 (polyclonal, E-AB-66298, Elabscince, 1:500), anti-caspase-1 (monoclonal, G6231-3D2, Sigma Aldrich, St. Louis, MO, USA, 1:500), anti-caspase-5 (monoclonal, MAA770Hu22, Cloud-clone Corp., Katy, TX, USA, 1:100), anti-GSDMD (monoclonal, E-AB-81473, Elabscience, 1:200), anti-IL-1β (polyclonal, ab2105, Abcam, Cambridge, MA, USA, 1:800), and anti-IL-18 (polyclonal, ab68435, Abcam, 1:2000).

Techniques: Immunohistochemistry

Journal: Scientific Reports

Article Title: Significance of elevated serum and hepatic NOD-like receptor pyrin domain containing 3 (NLRP3) in hepatitis C virus-related liver disease

doi: 10.1038/s41598-022-22022-5

Figure Lengend Snippet: Serum nucleotide-binding and oligomerization domain-like receptor pyrin-domain-containing 3 (NLRP3) level (pg/ml) in patients with HCV-related liver disease and healthy controls [Data are expressed as median (interquartile range), 1040 (395) pg/ml vs 695 (183) pg/ml respectively, χ 2 = 23.888, P < 0.001, the Mood’s median test].

Article Snippet: Liver sections were deparaffinized and incubated with the following primary antibodies at 4 °C overnight in a humid chamber: anti-NLRP3 antibody (CSB-PA015871LA01HU)—rabbit polyclonal antibody at a dilution 1:20 (CUSABIO, Wuhan, Hubei Province, China); and anti-CASP1 ( cleaved ) antibody (MBS301007)—rabbit polyclonal antibody at a dilution 1:50 (MyBioSource, San Diego, CA, USA).

Techniques: Binding Assay

Journal: Scientific Reports

Article Title: Significance of elevated serum and hepatic NOD-like receptor pyrin domain containing 3 (NLRP3) in hepatitis C virus-related liver disease

doi: 10.1038/s41598-022-22022-5

Figure Lengend Snippet: Immunohistochemical staining of liver tissues from patients with hepatitis C virus-related liver disease. ( A ) METAVIR stage F1 with mild activity (A1), and no steatosis. Moderate brown positive NLRP3 (nucleotide-binding and oligomerization domain-like receptor pyrin-domain-containing 3) staining of hepatocytes is seen (staining score 4, low expression) (anti-NLRP3 × 200); ( B ) High power view demonstrating brown cytoplasmic NLRP3 expression in hepatocytes (anti-NLRP3 × 400); ( C ) METAVIR stage F4 (cirrhosis) with moderate activity (A2), and steatosis. Strong NLRP3 expression is seen (staining score 12, high expression) (anti-NLRP3 × 200); ( D ) METAVIR stage F2, with moderate activity (A2). Weak brown positive Caspase-1 (CASP1) staining of hepatocytes is observed (staining score 1) (anti-CASP1 × 200). ( E ) METAVIR stage F3 with moderate activity (A2), and no steatosis. Moderate brown positive CASP1 staining of hepatocytes is seen (staining score 2) (anti-CASP1 × 200); and ( F ) METAVIR stage F4 (cirrhosis). Strong brown positive CASP1 staining of hepatocytes (staining score 3) (anti-CASP1 × 200).

Article Snippet: Liver sections were deparaffinized and incubated with the following primary antibodies at 4 °C overnight in a humid chamber: anti-NLRP3 antibody (CSB-PA015871LA01HU)—rabbit polyclonal antibody at a dilution 1:20 (CUSABIO, Wuhan, Hubei Province, China); and anti-CASP1 ( cleaved ) antibody (MBS301007)—rabbit polyclonal antibody at a dilution 1:50 (MyBioSource, San Diego, CA, USA).

Techniques: Immunohistochemical staining, Staining, Virus, Activity Assay, Binding Assay, Expressing

Journal: Scientific Reports

Article Title: Significance of elevated serum and hepatic NOD-like receptor pyrin domain containing 3 (NLRP3) in hepatitis C virus-related liver disease

doi: 10.1038/s41598-022-22022-5

Figure Lengend Snippet: Statistical correlations between ( A ) hepatic nucleotide-binding and oligomerization domain-like receptor pyrin-domain-containing 3 (NLRP3) and Caspase-1 (CASP1) staining scores (r = 0.810, P < 0.001), ( B ) serum NLRP3 levels (pg/ml) and hepatic NLRP3 final staining score (r = 0.749, P < 0.001), and ( C ) serum NLRP3 levels (pg/ml) and hepatic CASP1 staining score (r = 0.577, P = 0.001) in patients with hepatitis C virus-related liver disease (n = 34).

Article Snippet: Liver sections were deparaffinized and incubated with the following primary antibodies at 4 °C overnight in a humid chamber: anti-NLRP3 antibody (CSB-PA015871LA01HU)—rabbit polyclonal antibody at a dilution 1:20 (CUSABIO, Wuhan, Hubei Province, China); and anti-CASP1 ( cleaved ) antibody (MBS301007)—rabbit polyclonal antibody at a dilution 1:50 (MyBioSource, San Diego, CA, USA).

Techniques: Binding Assay, Staining, Virus

Journal: Scientific Reports

Article Title: Significance of elevated serum and hepatic NOD-like receptor pyrin domain containing 3 (NLRP3) in hepatitis C virus-related liver disease

doi: 10.1038/s41598-022-22022-5

Figure Lengend Snippet: Statistical correlations between serum nucleotide-binding and oligomerization domain-like receptor pyrin-domain-containing 3 (NLRP3) levels (pg/ml), hepatic NLRP3 expression, and other parameters in patients with hepatitis C virus (HCV)-related liver disease.

Article Snippet: Liver sections were deparaffinized and incubated with the following primary antibodies at 4 °C overnight in a humid chamber: anti-NLRP3 antibody (CSB-PA015871LA01HU)—rabbit polyclonal antibody at a dilution 1:20 (CUSABIO, Wuhan, Hubei Province, China); and anti-CASP1 ( cleaved ) antibody (MBS301007)—rabbit polyclonal antibody at a dilution 1:50 (MyBioSource, San Diego, CA, USA).

Techniques: Expressing, Virus, Staining, Activity Assay

Journal: Scientific Reports

Article Title: Significance of elevated serum and hepatic NOD-like receptor pyrin domain containing 3 (NLRP3) in hepatitis C virus-related liver disease

doi: 10.1038/s41598-022-22022-5

Figure Lengend Snippet: Serum levels (pg/ml) and hepatic expression of nucleotide-binding and oligomerization domain-like receptor pyrin-domain-containing 3 (NLRP3) in: ( A & B ) patients with severe necroinflammation (A3) vs patients with mild/moderate necroinflammation (A1-A2) ( P < 0.001 and P = 0.013 respectively), ( C & D ) patients with advanced fibrosis/cirrhosis (F3-F4) vs patients with early fibrosis (F1-F2) ( P < 0.001 for both), and ( E & F ) patients with significant steatosis (grade 2–3) vs patients with non-significant steatosis (grade 0–1) ( P < 0.001 and P = 0.001 respectively).

Article Snippet: Liver sections were deparaffinized and incubated with the following primary antibodies at 4 °C overnight in a humid chamber: anti-NLRP3 antibody (CSB-PA015871LA01HU)—rabbit polyclonal antibody at a dilution 1:20 (CUSABIO, Wuhan, Hubei Province, China); and anti-CASP1 ( cleaved ) antibody (MBS301007)—rabbit polyclonal antibody at a dilution 1:50 (MyBioSource, San Diego, CA, USA).

Techniques: Expressing, Binding Assay

Journal: Scientific Reports

Article Title: Significance of elevated serum and hepatic NOD-like receptor pyrin domain containing 3 (NLRP3) in hepatitis C virus-related liver disease

doi: 10.1038/s41598-022-22022-5

Figure Lengend Snippet: The diagnostic performance of serum nucleotide-binding and oligomerization domain-like receptor pyrin-domain-containing 3 (NLRP3) (pg/ml) in determining the severity of liver necroinflammation, fibrosis, and steatosis.

Article Snippet: Liver sections were deparaffinized and incubated with the following primary antibodies at 4 °C overnight in a humid chamber: anti-NLRP3 antibody (CSB-PA015871LA01HU)—rabbit polyclonal antibody at a dilution 1:20 (CUSABIO, Wuhan, Hubei Province, China); and anti-CASP1 ( cleaved ) antibody (MBS301007)—rabbit polyclonal antibody at a dilution 1:50 (MyBioSource, San Diego, CA, USA).

Techniques: Diagnostic Assay

Journal: Scientific Reports

Article Title: Significance of elevated serum and hepatic NOD-like receptor pyrin domain containing 3 (NLRP3) in hepatitis C virus-related liver disease

doi: 10.1038/s41598-022-22022-5

Figure Lengend Snippet: Receiver operating characteristic curve showing the sensitivity and specificity of ( A ) serum nucleotide-binding and oligomerization domain-like receptor pyrin-domain-containing 3 (NLRP3) (pg/ml), serum aspartate aminotransferase (AST) (U/L), and serum alanine aminotransferase (ALT) (U/L) in discriminating severe liver necroinflammation (A3) from mild/moderate liver necroinflammation (A1-A2) (area under the curve (AUC) = 0.951 vs 0.676, P = 0.032 and 0.692, P = 0.034 respectively), ( B ) serum NLRP3 (pg/ml), aspartate aminotransferase to platelet ratio index (APRI), and Fibrosis-4 index (FIB-4) in discriminating advanced liver fibrosis/cirrhosis (F3-F4) from early liver fibrosis (F1-F2) (AUC = 0.971 vs 0.754, P = 0.036 and 0.806, P = 0.075 respectively), ( C ) serum NLRP3 (pg/ml) in discriminating significant steatosis (grade 2–3) from non-significant steatosis (grade 0–1) (AUC = 0.917, P < 0.001).

Article Snippet: Liver sections were deparaffinized and incubated with the following primary antibodies at 4 °C overnight in a humid chamber: anti-NLRP3 antibody (CSB-PA015871LA01HU)—rabbit polyclonal antibody at a dilution 1:20 (CUSABIO, Wuhan, Hubei Province, China); and anti-CASP1 ( cleaved ) antibody (MBS301007)—rabbit polyclonal antibody at a dilution 1:50 (MyBioSource, San Diego, CA, USA).

Techniques: 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