Journal: PLoS ONE

Article Title: The ROS Scavenger, NAC, Regulates Hepatic Vα14 i NKT Cells Signaling during Fas mAb-Dependent Fulminant Liver Failure

doi: 10.1371/journal.pone.0038051

Figure Lengend Snippet: ( a ) Serum ALT levels of naïve WT mice, Fas mAb-treated WT and Jα18 −/− mice at 4.5 h. ( b–d ) H & E staining of liver sections from naïve WT mice, Fas mAb-treated WT and Jα18 −/− mice at 4.5 h. Livers from Fas mAb-treated WT mice ( c ) showed extensive damage with destruction of hepatocytes and distortion of normal liver architecture. The hepatocytes show hemorrhagic necrosis (white arrows) and characteristic signs of apoptosis (black arrows) including chromatin condensation and cellular shrinkage. In comparison, livers from Fas mAb-treated Jα18 −/− mice showed only minimal damage and retained the normal architecture ( d ). Liver from a naïve WT mouse is illustrated in ( b ) for comparison. ( e ) Western blot analysis of active caspase 3, T-bet, pSTAT-1, nitrotyrosine and GAPDH expression in the liver of PBS-treated WT mice and agonistic Fas mAb-treated WT and Jα18 −/− mice at 4.5 h. ( f ) TUNEL staining of liver sections from WT and Jα18 −/− mice at 4.5 h after Fas mAb injection in which WT mice showed intense TUNEL staining characteristic of apoptosis whereas Jα18 −/− mice showed less/reduced TUNEL staining. ( g ) HPLC measurement of hepatic GSH levels in PBS-treated WT mice and Fas mAb-treated WT and Jα18 −/− mice at 4.5 h. in a and g are presented as mean ± s.e.m with n = 5 mice/group; * P <0.05 by one-way analysis of variance followed by Newman-Kuels post hoc test. All experiments were conducted twice.

Article Snippet: Active caspase 3 Ab (clone 269518) was obtained from R & D systems (Minneapolis, MN) whereas pSTAT-1 (Tyr701) Ab was supplied by BD Pharmingen.

Techniques: Staining, Western Blot, Expressing, TUNEL Assay, Injection

Journal: PLoS ONE

Article Title: The ROS Scavenger, NAC, Regulates Hepatic Vα14 i NKT Cells Signaling during Fas mAb-Dependent Fulminant Liver Failure

doi: 10.1371/journal.pone.0038051

Figure Lengend Snippet: ( a ) Serum ALT levels in WT and IFN-γ −/− mice after PBS or NAC treatment during agonistic Fas mAb-induced FLF. ( b ) H & E staining of liver sections of WT and IFN-γ −/− mice after PBS or NAC treatment during agonistic Fas mAb-induced FLF. As shown in top panel , livers from Fas mAb-treated WT and IFN-γ −/− mice displayed widespread hepatocyte damage including hemorrhagic necrosis (white arrows) and apoptosis (black arrows) that distorted normal liver architecture. In contrast, liver sections of WT and IFN-γ −/− mice treated with NAC during Fas mAb-induced FLF ( bottom panel ) showed reduced hepatocyte damage and retained near normal architecture. ( c & e ) Western blot analysis of hepatic active caspase 3, T-bet, pSTAT-1 expression levels and nitrotyrosine formation in WT and IFN-γ −/− mice after PBS or NAC treatment during Fas mAb-induced FLF. ( d ) TUNEL staining of liver sections from WT and IFN-γ −/− mice treated with PBS during Fas mAb-induced FLF showed intense TUNEL staining characteristic of apoptosis whereas WT and IFN-γ −/− mice treated with NAC mice showed minimal TUNEL staining. in a are presented as mean ± s.e.m with n = 3–6 mice/group. * P <0.05, ≠ P <0.05 by one-way analysis of variance followed by Newman-Kuels post hoc test. All experiments were performed twice.

Article Snippet: Active caspase 3 Ab (clone 269518) was obtained from R & D systems (Minneapolis, MN) whereas pSTAT-1 (Tyr701) Ab was supplied by BD Pharmingen.

Techniques: Staining, Western Blot, Expressing, TUNEL Assay

Journal: PLoS ONE

Article Title: The ROS Scavenger, NAC, Regulates Hepatic Vα14 i NKT Cells Signaling during Fas mAb-Dependent Fulminant Liver Failure

doi: 10.1371/journal.pone.0038051

Figure Lengend Snippet: An endogenous mediator inhibited by NAC (possibly ROS) mediates Fas mAb-dependent FLF by promoting intrahepatic Vα14 i NKT cells signaling, upregulation of pSTAT-1 and pSTAT-1-regulated genes, caspase 3 and T-bet, induction of hepatocyte damage and fatal/lethal immunopathological events in the liver that ultimately leads to FLF.

Article Snippet: Active caspase 3 Ab (clone 269518) was obtained from R & D systems (Minneapolis, MN) whereas pSTAT-1 (Tyr701) Ab was supplied by BD Pharmingen.

Techniques:

Journal: Frontiers in Medicine

Article Title: Ross River Virus Immune Evasion Strategies and the Relevance to Post-viral Fatigue, and Myalgic Encephalomyelitis Onset

doi: 10.3389/fmed.2021.662513

Figure Lengend Snippet: Examples of ADE-mediated virus infections and the impact on subsequent cellular pathways and cytokine expression.

Article Snippet: Flaviviridae ( Flavivirus ) , Dengue - ( Fever, Shock, Myalgia, Hemorrhage ) , Monocyte, Macrophage - Post ADE suppression of IL-12, IFN-γ TNF; Increased IL-6, IL-10 (0-5 days p.i) with pSTAT-1, IRF-1 impacted; Increased IL-10 expression SOCS3, Syk-regulated; Early NOS2 via RLR-MAVS (without IFN); Autophagy role (ATG5) in IFN restriction; Early Syk - ERK1/2 IL-1β stimulation independent of DEN replication. Mast cell/Basophil - Post-ADE (72 hrs) significant increases for IL-1β, IL-6, not GM-CSF , ( ) ( ) ( ) ( ) ( ) ( ) ( ) .

Techniques: Virus, Expressing, Phospho-proteomics, Infection

Journal: Frontiers in Immunology

Article Title: Pellino-1 Regulates Immune Responses to Haemophilus influenzae in Models of Inflammatory Lung Disease

doi: 10.3389/fimmu.2019.01721

Figure Lengend Snippet: Pellino-1 regulates macrophage STAT-1 signaling. MDMs were transfected with siRNA targeting Pellino-1 or scrambled siRNA (control). (A,B,D) 48 h following transfection, cells were stimulated with IFN-γ [20 ng/ml], IL-4 [20 ng/ml], IL-10 [20 ng/ml] or LPS [1 ng/ml] for 24 h. Whole cell lysates were analyzed by Western blot using antibodies specific to STAT-1, pSTAT-6, Pellino-1 or actin for 4 independent donors (A) . pSTAT-1 bands for IFN-γ-treated cells were analyzed by densitometry (B) . In separate experiments cells were challenged with NTHi (MOI 1 and 10) for 24 h and lysates probed for Pellino-1 and actin (C) . RNA was purified from cell lysates and transcribed to cDNA. CD206 gene product was measured by qPCR (D) . Fold changes are shown relative to media treated MDMs transfected with scrambled siRNA (first open bar). Data are presented as mean ± SEM of n = 4 independent experiments. Significant differences are indicated by ** P < 0.01.

Article Snippet: Membranes were probed against antibodies to Pellino-1, actin (Santa Cruz, Santa Cruz, CA), pStat-1 or Stat-6 (Cell-Signaling Technology, Leiden, The Netherlands).

Techniques: Transfection, Western Blot, Purification