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: medRxiv

Article Title: Polymorphism in IFNAR contributes to glucocorticoid response and outcome in ARDS and COVID-19

doi: 10.1101/2022.03.10.22272123

Figure Lengend Snippet: (A ) STAT1 expression in the lung after 4-day culture in the presence of IFN beta with or without hydrocortisone (HC). ( B) pSTAT1 expression in the same specimens as in A. ( C) Example photomicrographs showing higher STAT2 expression in a TT patient than in a CT patient and the effect of HC on its nuclear translocation. Most STAT2 remains in the cytoplasm of the CT patients, whereas nuclear expression is prominent in the TT patient. Indicated insets are shown in the bottom row. Arrows. ( D ) Combined results of all patients noting that two CT samples are excluded in the data as the patients were already under glucocorticoid treatment at the time of sample acquisition. Ns, not significant; *P<0.05; **P<0.01; and ***P<0.001

Article Snippet: The first stage antibodies were anti-alpha chain of the IFN alpha/beta receptor (St John’s Laboratory STJ112765) that was used 1:2000 and 1:5000 and anti-Stat1 (1:400, 9175S), anti-pStat1(1:100, 9167S) and anti-Stat2 (1:200, 72604S) all from Cell Signalling.

Techniques: Expressing, Translocation Assay

Journal: Microbiology Spectrum

Article Title: Condition-dependent effects of Elexacaftor/Tezacaftor/Ivacaftor (Trikafta) on Aspergillus fumigatus growth

doi: 10.1128/spectrum.02275-24

Figure Lengend Snippet: High concentration of CFTR modulators (Elexacaftor [VX445], Tezacaftor [VX661], and Ivacaftor [VX770]) impairs macrophage control of A. fumigatus growth. ( A ) Measurement of A. fumigatus fungal length (Strain: DAL-DSred) infecting bone marrow-derived macrophages (BMDM) at a multiplicity of infection (MOI) of 5, using the IncuCyte live-cell analysis system in the presence of 3VX (VX445/VX661/VX770) treatment at [3VX] S , [3VX] E , or [3VX] 10μM , as well as with [Veh]. * P < 0.05, ** P < 0.01 (two-way ANOVA with Dunnett’s multiple comparisons test); data are presented as mean ± SEM, n = 3. ( B ) Release of TNF⍺ in BMDMs was assessed after 20 h with A. fumigatus (MOI of 15). ** P < 0.01 (one-way ANOVA with Dunnett’s multiple comparisons test); data are presented as mean ± SEM, n = 4. ( C ) Normalized phagocytosis to vehicle control ([Veh]) at 20 h time point. **P < 0.01 , ****P < 0.005 (one sample t test, theoretical mean of 1); data are presented as mean ± SEM, n = 4. ( D ) Representative measurement of zymosan phagocytosis by primed BMDMs in the presence of 3VX treatment at concentrations [3VX] S , [3VX] E or [3VX] 10μM , as well as in the presence of cytochalasine D or [Veh]. ( E ) Quantification of viable conidia following 4 h of BMDM infection in the presence of 3VX treatment at [3VX] S , [3VX] E , or [3VX] 10μM , as well as with [Veh]. ( F ) Immunoblot analysis of pro-caspase-1 ( P45 ), the caspase-1 subunit p20 ( P20 ), and β-actin of primed BMDMs left untreated (medium alone [Med]) or in the presence of 3VX treatment at [3VX] S , [3VX] E , or [3VX] 10μM , as well as with [Veh] during 20 h after infection with A. fumigatus (MOI of 15). ( G ) Release of IL-1β in BMDMs assessed after 20 h infection with A. fumigatus (MOI of 15) left untreated (medium alone [Med]) or in the presence of 3VX treatment at [3VX] S , [3VX] E , or [3VX] 10μM , as well as with [Veh]. *P < 0.05 , **P < 0.01 (one-way ANOVA with Dunnett’s multiple comparisons test); data are presented as mean ± SEM, n = 3.

Article Snippet: Cytokine levels were determined by ELISA using kits according to the manufacturer’s instructions: TNF⍺ DuoSet ELISA (DY410, R&D Systems) and IL-1β (DY401, R&D Systems).

Techniques: Concentration Assay, Control, Derivative Assay, Infection, Cell Analysis, Western Blot

Journal: Journal of Orthopaedic Translation

Article Title: The CD163/TWEAK/Fn14 axis: A potential therapeutic target for alleviating inflammatory bone loss

doi: 10.1016/j.jot.2024.09.002

Figure Lengend Snippet: TWEAK mediates the phenotype of osteoporosis in CD163 −/− mice . (A, B) Representative images and quantification of TWEAK from CD163 −/− male mice or their WT male littermates. Scale bar: 50 μm (top), 10 μm (bottom). (C, D) Representative immunoblotting and quantification of CD163 and TWEAK from CD163 −/− male mice or their WT male littermates. (E) Quantification of sTWEAK level in the peripheral circulation using ELISA. (F, G) Representative images and quantification of TWEAK from mice treated with or without LPS. Scale bar: 50 μm (top), 10 μm (bottom). (H, I) Representative images and quantification for proportion of TWEAK + cells in BMCs from CD163 −/− male mice or their WT male littermates. The numbers in the upper right corner represent the average proportion of TWEAK + cells. (J) Representative images of TWEAK + (red), CD68 + (green), and F4/80 + (green) cells from CD163 −/− mice. White arrow heads: the CD68-and TWEAK- double positive cells. Nuclei were stained with DAPI (blue). Scale: 50 μm (left), 10 μm (right). (K, L) Representative μCT images and quantification of femora from CD163 −/− mice treated with or without MTW-1. (M, N) Representative μCT images and quantification of femora from WT mice treated with 0.1%BSA (Vehicle), rTWEAK, or rTWEAK plus rCD163. Dots represent individual mice. The data are presented as mean ± s. e.m., ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The concentration of sTWEAK in serum samples were measured using a commercial mouse TWEAK ELISA kit (EL023987MO, CUSABIO, Wuhan, China) according to the manufacturer's directions.

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Staining