Journal: iScience

Article Title: Chronic HDM exposure shows time-of-day and sex-based differences in inflammatory response associated with lung circadian clock disruption

doi: 10.1016/j.isci.2023.107580

Figure Lengend Snippet: Serum levels of total and HDM-specific immunoglobulins show a time-of-day response and sex-based differences to chronic HDM exposure Total IgE, Total IgG, HDM-specific IgE, HDM-specific IgG, HDM-specific IgG1, HDM-specific IgG2b, HDM-specific IgM, and HDM-specific IgA in the serum of chronic PBS- and HDM-exposed females and males at ZT0 and ZT12 were determined by ELISA. Data were expressed as ng/ml and mg/ml for total IgE and total IgG, respectively. HDM-specific immunoglobulins (IgE, IgG, IgG1, IgG2b, IgM, and IgA) in the serum were determined by commercially available ELISA kits (Chondrex, Inc.). Unable to detect HDM-specific IgG2a and IgG3 levels in PBS- and HDM-exposed mice. Data for the HDM-specific immunoglobulins were expressed as absorbance at 450 nm or μg/ml. Data are shown as mean ± SEM, Two-way ANOVA followed by Tukey’s multiple comparison test (n = 5/group). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, compared to respective control (PBS) at ZT0 or ZT12; # p < 0.05, compared to HDM at ZT0 vs. ZT12; # # p < 0.01, compared to HDM at ZT0 vs. ZT12. Summary statistics for interaction between Treatment x Sex x Time were analyzed using generalized linear modeling using R (see Table S2 ).

Article Snippet: Mouse Anti-House Dust Mite (HDM) IgG2b Antibody Assay Kit , Chondrex, Inc. , Cat# 3035.

Techniques: Enzyme-linked Immunosorbent Assay, Comparison, Control

Journal: iScience

Article Title: Chronic HDM exposure shows time-of-day and sex-based differences in inflammatory response associated with lung circadian clock disruption

doi: 10.1016/j.isci.2023.107580

Figure Lengend Snippet:

Article Snippet: Mouse Anti-House Dust Mite (HDM) IgG2b Antibody Assay Kit , Chondrex, Inc. , Cat# 3035.

Techniques: Purification, Recombinant, Staining, Enzyme-linked Immunosorbent Assay, Software

Journal: Toxicological Sciences

Article Title: Natural Killer Cells Mediate Severe Liver Injury in a Murine Model of Halothane Hepatitis

doi: 10.1093/toxsci/kfr005

Figure Lengend Snippet: HAL hepatotoxicity and the induced inflammatory response depend on ovarian hormones. (A) For each mouse, the stage of estrous cycle was determined by vaginal cytology analysis before treatment with HAL (5 mmol/kg, ip). Plasma ALT activity was measured 12 h after HAL administration (n = 3–5 per group). P, proestrus; E/M, estrus/metestrus; D, diestrus. *significantly different from other groups. (B) Plasma ALT activity was evaluated 12 h after vehicle (VEH) or HAL (15 mmol/kg, ip) administration in OVX or SHAM mice (n = 3–5 per group). *significantly different from respective VEH control; #significantly different from HAL-treated SHAM mice. (C) Plasma IFN-γ concentration was evaluated in at various times after HAL administration (n = 5–6 per group). #significantly different from time-matched male group; *significantly different from sex-matched 6 h group. (D) IFN-γ concentration was evaluated 12 h after HAL treatment in SHAM and OVX mice (n = 4 per group). *significantly different from SHAM group.

Article Snippet: The plasma concentration of IFN-γ was measured using a BD OpEIA mouse IFN-γ ELISA kit purchased from BD Biosciences (San Diego, CA).

Techniques: Activity Assay, Concentration Assay

Journal: Toxicological Sciences

Article Title: Natural Killer Cells Mediate Severe Liver Injury in a Murine Model of Halothane Hepatitis

doi: 10.1093/toxsci/kfr005

Figure Lengend Snippet: IFN-γ KO mice are protected from developing severe HAL hepatotoxicity. Female WT BALB/cJ (WT) and IFN-γ KO mice were treated with HAL (15 mmol/kg, ip), and plasma and liver samples were collected at various times. (A) Plasma ALT activity was evaluated 8 and 12 h after HAL treatment (n = 5–6 per group). *significantly different from HAL-treated WT mice. (B) Immunoblot detection of TFA protein adducts in liver homogenates (n = 3 per group). (C) Hematoxylin and eosin liver sections from HAL-treated WT and IFN-γ KO mice 30 h after treatment. Labeled in picture are central vein (CV) and portal triad (PT). Images were photographed at ×200 magnification.

Article Snippet: The plasma concentration of IFN-γ was measured using a BD OpEIA mouse IFN-γ ELISA kit purchased from BD Biosciences (San Diego, CA).

Techniques: Activity Assay, Western Blot, Labeling

Journal: Toxicological Sciences

Article Title: Natural Killer Cells Mediate Severe Liver Injury in a Murine Model of Halothane Hepatitis

doi: 10.1093/toxsci/kfr005

Figure Lengend Snippet: HMGB-1 release and the response to HAL in Tlr4Lps-d mice. (A) Plasma HMGB-1 concentration was evaluated at various times after HAL treatment (15 mmol/kg, ip) in male and female mice (n = 6 per group). VEH-treated animals had plasma HMGB1 concentrations < 5 pg/ml. #significantly different from sex-matched 6 h time point. *significantly different from time-matched male and all other female groups. (B and C) Female WT BALB/cBYJ (WT) mice and Tlr4Lps-d mice were treated with HAL (15 mmol/kg, ip). Plasma ALT activity and IFN-γ concentration were evaluated 24 h after HAL treatment (n = 4–5 per group). *significantly different from WT controls.

Article Snippet: The plasma concentration of IFN-γ was measured using a BD OpEIA mouse IFN-γ ELISA kit purchased from BD Biosciences (San Diego, CA).

Techniques: Concentration Assay, Activity Assay

Journal: Toxicological Sciences

Article Title: Natural Killer Cells Mediate Severe Liver Injury in a Murine Model of Halothane Hepatitis

doi: 10.1093/toxsci/kfr005

Figure Lengend Snippet: KC-depleted mice and CD1d KO and RAGNULL mice develop severe HAL-induced liver injury. Control- or clodronate liposome-pretreated mice were given VEH or HAL (15 mmol/kg, ip), and plasma and liver samples were collected 24 h later. Plasma ALT activity (A) and IFN-γ concentration (B) were evaluated (n = 4–6 per group). WT BALB/cJ (WT), NKT-deficient mice (CD1d KO), or T- and B cell–deficient mice (RAGNULL) were treated with HAL (15 mmol/kg, ip). (C) Plasma ALT activity was evaluated in WT and CD1d KO mice 12 and 24 h after HAL administration (n = 5 per group). (D) Plasma ALT activity was evaluated in HAL-treated WT and RAGNULL mice 12 h after HAL administration (n = 5 per group). (E and F) Hematoxylin and eosin liver sections taken 24 h after HAL treatment of CD1d KO and RAGNULL mice. Labeled in picture are central vein (CV) and portal triad (PT). Arrowheads identify areas of necrosis. Images were photographed at ×200X magnification.

Article Snippet: The plasma concentration of IFN-γ was measured using a BD OpEIA mouse IFN-γ ELISA kit purchased from BD Biosciences (San Diego, CA).

Techniques: Activity Assay, Concentration Assay, Labeling

Journal: Toxicological Sciences

Article Title: Natural Killer Cells Mediate Severe Liver Injury in a Murine Model of Halothane Hepatitis

doi: 10.1093/toxsci/kfr005

Figure Lengend Snippet: HAL-induced hepatotoxicity depends on NK cell activity. Mice treated with IgG or anti-AsGM1 were given HAL (15 mmol/kg, ip) as described in Methods section, and plasma samples were collected at 12 and 24 h. Plasma ALT activity (A) and IFN-γ concentration (B) (n = 4–6 per group). *significantly different from time-matched controls. Plasma ALT activity (C) and IFN-γ concentration (D) in WT and BALBPrf1 mice 12 h after HAL administration (n = 4 per group). *significantly different from WT mice.

Article Snippet: The plasma concentration of IFN-γ was measured using a BD OpEIA mouse IFN-γ ELISA kit purchased from BD Biosciences (San Diego, CA).

Techniques: Activity Assay, Concentration Assay

Journal: Toxicological Sciences

Article Title: Natural Killer Cells Mediate Severe Liver Injury in a Murine Model of Halothane Hepatitis

doi: 10.1093/toxsci/kfr005

Figure Lengend Snippet: Proposed mechanism of innate immune-mediated severe HAL-induced liver injury. (A) In the absence of stress stimulation, self-proteins such as H2Dd are expressed on the plasma membrane of hepatocytes and RAE-1 is not; this condition keeps NK cells quiescent. (B) When hepatocytes are exposed to HAL, intracellular TFA adducts form (1). This induces a stress response in hepatocytes (2) that alters the surface NK receptor ligands (3) and activates NK cells. Activated NK cells release IFN-γ as well as the contents of cytotoxic granules (4), such as perforin and granzyme B, which contribute to hepatocellular necrosis (5). Damaged hepatocytes release endogenous danger signals, such as HMGB-1 (6). These endogenous danger signals are ligands for TLR4 (7), and the resultant signals are involved in a positive feedback loop that further activates NK cells as well as recruits polymorphonuclear leukocytes (PMNs) (Scaffidi et al., 2002) that participate (8) in the progression of injury (Dugan et al., 2010).

Article Snippet: The plasma concentration of IFN-γ was measured using a BD OpEIA mouse IFN-γ ELISA kit purchased from BD Biosciences (San Diego, CA).

Techniques:

Journal: Cell metabolism

Article Title: Low-Dose Sorafenib Acts as a Mitochondrial Uncoupler and Ameliorates Nonalcoholic Steatohepatitis

doi: 10.1016/j.cmet.2020.04.011

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse TNF ELISA kit , Cloud-Clone Co. , Cat#SEA133Mu.

Techniques: Recombinant, Protease Inhibitor, Western Blot, SYBR Green Assay, Modification, Bicinchoninic Acid Protein Assay, Cholesterol Assay, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Gene Knockout, Knock-Out, Plasmid Preparation, Software

Journal: Cell metabolism

Article Title: Low-Dose Sorafenib Acts as a Mitochondrial Uncoupler and Ameliorates Nonalcoholic Steatohepatitis

doi: 10.1016/j.cmet.2020.04.011

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse TNF ELISA kit , Cloud-Clone Co. , Cat#SEA133Mu.

Techniques: Recombinant, Protease Inhibitor, Western Blot, SYBR Green Assay, Modification, Bicinchoninic Acid Protein Assay, Cholesterol Assay, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Gene Knockout, Knock-Out, Plasmid Preparation, Software

Journal: Cell Reports Medicine

Article Title: A CSF-1R-blocking antibody/IL-10 fusion protein increases anti-tumor immunity by effectuating tumor-resident CD8 + T cells

doi: 10.1016/j.xcrm.2023.101154

Figure Lengend Snippet: Transcriptomic and TCR repertoire analysis of the samples from mice treated with BF10 versus its subcomponents (A) An illustration of mice receiving the indicated treatment followed by tumor isolation for bulk RNA-seq and gene set enrichment analysis (GSEA). (B–F) Enrichment plots of genes sets in BF10 versus control in mouse tumor samples. Significant pathways were identified by GSEA gene set of IFN-γ response (B), IFN-α response (C), inflammatory response (D), complement (E), and genes upregulated in CD8 T cells (GEO: GSE41867 ) (F). (G) The heatmap of GSEA enriched pathways in treatment groups. The normalized enrichment score (NES) and p value are shown. (H) Schema of bioinformatic strategy to study the effect of BF10 within tumor microenvironment. Tumors derived from two syngeneic tumor models (HNSC/Q1-2 and BRCA/4T1) treated with BF10, IL-10-Fc, αCSF-1R, or control were collected for bulk RNA-seq to investigate the differentially expressed genes (DEGs; >1.5-fold and p < 0.05 or <0.5-fold and p < 0.05). DEGs were analyzed using 2 modules: (1) Bioinformatics Database for Annotation, Visualization, and Integrated Discovery (DAVID; https://david.ncifcrf.gov/ ) and (2) Ingenuity Pathway Analysis (IPA). (I) DAVID functional Gene Ontology analysis. (J) IPA. (K) Immune repertoire TCR sequencing of CD8 + T cells. Tumor-bearing mice received treatments of Ctrl-IgG, IL-10, anti-CSF1R, or BF10 for 3 doses, followed by RNA extraction of isolated CD8 + T cells for TCR immune repertoire analysis. The data are shown as clonotype diversity and distribution of both TRAC and TRBC CDR3 sequencing from tumor and spleen. The number of the bracket indicates observed diversity in the enrichment metrics of QIAseq-RNA Immune Repertoire Application. (L) Examination of the TCR immune repertoire using Morisita-Horn index.

Article Snippet: The same amount RNA (100 ng and RIN >8) of indicated group was applied to construct RNA-seq libraries using QIAseq Mouse TCR Sequencing kit (#333705, Qiagen).

Techniques: Isolation, RNA Sequencing, Control, Derivative Assay, Functional Assay, Sequencing, RNA Extraction

Journal: Cell Reports Medicine

Article Title: A CSF-1R-blocking antibody/IL-10 fusion protein increases anti-tumor immunity by effectuating tumor-resident CD8 + T cells

doi: 10.1016/j.xcrm.2023.101154

Figure Lengend Snippet:

Article Snippet: The same amount RNA (100 ng and RIN >8) of indicated group was applied to construct RNA-seq libraries using QIAseq Mouse TCR Sequencing kit (#333705, Qiagen).

Techniques: Control, Flow Cytometry, RNA Sequencing, Sequencing, Recombinant, Labeling, Enzyme-linked Immunosorbent Assay, Binding Assay, Polymer, Cell Isolation, Selection, Software