Cxcl8b and Cxcr2 Regulate Neutrophil Migration through Bloodstream in Zebrafish

Neutrophils play an essential role during an inflammatory response, which is dependent on their rapid recruitment from the bone marrow to the vasculature. However, there is no information about the molecular signals that regulate neutrophil entry to circulation during an inflammatory process in humans. This is mainly due to the lack of a suitable model of study that contains similar set of molecules and that allows in vivo analyses. In this study, we used the zebrafish to assess the role of Cxcl8a, Cxcl8b, and Cxcr2 in neutrophil migration to blood circulation after injury. Using Tg(BACmpx:GFP)i114 transgenic embryos and two damage models (severe and mild), we developed in vivo lack of function assays. We found that the transcription levels of cxcl8a, cxcl8b, and cxcr2 were upregulated in the severe damage model. In contrast, only cxcr2 and cxcl8a mRNA levels were increased during mild damage. After knocking down Cxcl8a, neutrophil quantity decreased at the injury site, while Cxcl8b decreased neutrophils in circulation. When inhibiting Cxcr2, we observed a decrease in neutrophil entry to the bloodstream. In conclusion, we identified different functions for both Cxcl8 paralogues, being the Cxcl8b/Cxcr2 axis that regulates neutrophil entry to the bloodstream, while Cxcl8a/Cxcr2 regulates the migration to the affected area.

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Involvement of the Nonneuronal Cholinergic System in Bone Remodeling in Rat Midpalatal Suture after Rapid Maxillary Expansion

BioMed Research International ◽

10.1155/2016/8106067 ◽

2016 ◽

Vol 2016 ◽

pp. 1-8 ◽

Cited By ~ 2

Author(s):

Xiaoxia Che ◽

Jie Guo ◽

Lue Wang ◽

Cong Miao ◽

Lihua Ge ◽

...

Keyword(s):

Bone Formation ◽

Bone Remodeling ◽

Cholinergic System ◽

Dominant Role ◽

Suitable Model ◽

Mrna Levels ◽

Maxillary Expansion ◽

Expansion Model

Few studies sought to analyze the expression and function of the nonneuronal acetylcholine system in bone remodelingin vivodue to the lack of suitable models. We established a rat maxilla expansion model in which the midline palatine suture of the rat was rapidly expanded under mechanical force application, inducing tissue remodeling and new bone formation, which could be a suitable model to investigate the role of the nonneuronal acetylcholine system in bone remodelingin vivo.During the expansion, the expression pattern changes of the nonneuronal cholinergic system components and the mRNA levels of OPG/RANKL were detected by immunohistochemistry or real-time PCR. The value of the RANKL/OPG ratio significantly increased after 1 day of expansion, indicating dominant bone resorption induced by the mechanical stimulation; however after 3 days of expansion, the value of the RANKL/OPG ratio significantly decreased, suggesting a dominant role of the subsequent bone formation process. Increasing expression of Ach was detected after 3 days of expansion which indicated that ACh might play a role in bone formation. The mRNA expression levels of other components also showed observable changes during the expansion which confirmed the involvement of the nonneuronal cholinergic system in the process of bone remodelingin vivo. Further researches are still needed to figure out the detailed functions of the nonneuronal cholinergic system and its components.

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A Yes-Associated Protein (YAP) and Insulin-Like Growth Factor 1 Receptor (IGF-1R) Signaling Loop Is Involved in Sorafenib Resistance in Hepatocellular Carcinoma

Cancers ◽

10.3390/cancers13153812 ◽

2021 ◽

Vol 13 (15) ◽

pp. 3812

Author(s):

Mai-Huong T. Ngo ◽

Sue-Wei Peng ◽

Yung-Che Kuo ◽

Chun-Yen Lin ◽

Ming-Heng Wu ◽

...

Keyword(s):

Nuclear Translocation ◽

Combination Index ◽

Specific Inhibitor ◽

In Vivo Model ◽

Mrna Levels ◽

Margin Distribution ◽

Related Proteins ◽

Emt Markers

The role of a YAP-IGF-1R signaling loop in HCC resistance to sorafenib remains unknown. Method: Sorafenib-resistant cells were generated by treating naïve cells (HepG2215 and Hep3B) with sorafenib. Different cancer cell lines from databases were analyzed through the ONCOMINE web server. BIOSTORM–LIHC patient tissues (46 nonresponders and 21 responders to sorafenib) were used to compare YAP mRNA levels. The HepG2215_R-derived xenograft in SCID mice was used as an in vivo model. HCC tissues from a patient with sorafenib failure were used to examine differences in YAP and IGF-R signaling. Results: Positive associations exist among the levels of YAP, IGF-1R, and EMT markers in HCC tissues and the levels of these proteins increased with sorafenib failure, with a trend of tumor-margin distribution in vivo. Blocking YAP downregulated IGF-1R signaling-related proteins, while IGF-1/2 treatment enhanced the nuclear translocation of YAP in HCC cells through PI3K-mTOR regulation. The combination of YAP-specific inhibitor verteporfin (VP) and sorafenib effectively decreased cell viability in a synergistic manner, evidenced by the combination index (CI). Conclusion: A YAP-IGF-1R signaling loop may play a role in HCC sorafenib resistance and could provide novel potential targets for combination therapy with sorafenib to overcome drug resistance in HCC.

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Quantitative assessment of CYP11B1 and CYP11B2 expression in aldosterone-producing adenomas

Acta Endocrinologica ◽

10.1530/eje.0.1470795 ◽

2002 ◽

pp. 795-802 ◽

Cited By ~ 35

Author(s):

F Fallo ◽

V Pezzi ◽

L Barzon ◽

P Mulatero ◽

F Veglio ◽

...

Keyword(s):

Real Time ◽

Secretory Activity ◽

Zona Glomerulosa ◽

Zona Fasciculata ◽

Mrna Levels ◽

Rt Pcr ◽

Pathophysiological Role ◽

Aldosterone Production

BACKGROUND: The presence and pathophysiological role of CYP11B1 (11beta-hydroxylase) gene in the zona glomerulosa of human adrenal cortex is still controversial. METHODS: In order to specifically quantify CYP11B1, CYP11B2 (aldosterone synthase) and CYP17(17alpha-hydroxylase) mRNA levels, we developed a real-time RT-PCR assay and examined the expression in a series of adrenal tIssues, including six normal adrenals from patients adrenalectomized for renal cancer and twelve aldosterone-producing adenomas (APA) from patients with primary aldosteronism. RESULTS: CYP11B1 mRNA levels were clearly detected in normal adrenals, which comprised both zona glomerulosa and fasciculata/reticularis cells, but were also measured at a lower range (P<0.05) in APA. The levels of CYP11B2 mRNA were lower (P<0.005) in normal adrenals than in APA. CYP17 mRNAlevels were similar in normal adrenals and in APA. In patients with APA, CYP11B2 and CYP11B1 mRNA levels were not correlated either with basal aldosterone or with the change from basal aldosterone in response to posture or to dexamethasone. No correlation between CYP11B1 mRNA or CYP11B2 mRNA and the percentage of zona fasciculata-like cells was observed in APA. CONCLUSIONS: Real-time RT-PCR can be reliably used to quantify CYP11B1 and CYP11B2 mRNA levels in adrenal tIssues. Expression of CYP11B1 in hyperfunctioning zona glomerulosa suggests an additional formation of corticosterone via 11beta-hydroxylase, providing further substrate for aldosterone biosynthesis. CYP11B1 and CYP11B2 mRNA levels in APA are not related to the in vivo secretory activity of glomerulosa cells, where post-transcriptional factors might ultimately regulate aldosterone production.

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Targeted disruption of the murine junD gene results in multiple defects in male reproductive function

Development ◽

10.1242/dev.127.1.143 ◽

2000 ◽

Vol 127 (1) ◽

pp. 143-153 ◽

Cited By ~ 2

Author(s):

D. Thepot ◽

J.B. Weitzman ◽

J. Barra ◽

D. Segretain ◽

M.G. Stinnakre ◽

...

Keyword(s):

Reproductive Function ◽

Postnatal Growth ◽

Mrna Levels ◽

Targeted Disruption ◽

Male Reproductive Function ◽

Developing Heart ◽

Redundant Functions ◽

Transcription Factor Complex

JunD is one of three mammalian Jun proteins that contribute to the AP-1 transcription factor complex. Distinct regulation and functions have been proposed for each Jun member, but less is known about the biological functions of each of these proteins in vivo. To investigate the role of JunD, we have inactivated the murine gene by replacement with a bacterial lacZ reporter gene. Embryonic JunD expression was initially detected in the developing heart and cardiovascular system. Subsequent broadening phases of JunD expression were observed during embryonic development and expression in the adult was widespread in many tissues and cell lineages. Mutant animals lack JunD mRNA and protein and showed no evidence of upregulation of c-Jun and JunB mRNA levels. In contrast to the other two Jun members, homozygous JunD−/− mutant animals were viable and appeared healthy. However, homozygous JunD−/− animals showed a reduced postnatal growth. Furthermore, JunD−/− males exhibited multiple age-dependent defects in reproduction, hormone imbalance and impaired spermatogenesis with abnormalities in head and flagellum sperm structures. No defects in fertility were observed in JunD−/− female animals. These results provide evidence for redundant functions for members of the Jun family during development and specific functions for JunD in male reproductive function.

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Proof-of-concept clinical trial of etokimab shows a key role for IL-33 in atopic dermatitis pathogenesis

Science Translational Medicine ◽

10.1126/scitranslmed.aax2945 ◽

2019 ◽

Vol 11 (515) ◽

pp. eaax2945 ◽

Cited By ~ 29

Author(s):

Yi-Ling Chen ◽

Danuta Gutowska-Owsiak ◽

Clare S. Hardman ◽

Melanie Westmoreland ◽

Teena MacKenzie ◽

...

Keyword(s):

Atopic Dermatitis ◽

Therapeutic Potential ◽

Neutrophil Migration ◽

Neutrophil Infiltration ◽

Severity Index ◽

Fundamental Biology ◽

Targeted Inhibition

Targeted inhibition of cytokine pathways provides opportunities to understand fundamental biology in vivo in humans. The IL-33 pathway has been implicated in the pathogenesis of atopy through genetic and functional associations. We investigated the role of IL-33 inhibition in a first-in-class phase 2a study of etokimab (ANB020), an IgG1 anti–IL-33 monoclonal antibody, in patients with atopic dermatitis (AD). Twelve adult patients with moderate to severe AD received a single systemic administration of etokimab. Rapid and sustained clinical benefit was observed, with 83% achieving Eczema Area and Severity Index 50 (EASI50), and 33% EASI75, with reduction in peripheral eosinophils at day 29 after administration. We noted significant reduction in skin neutrophil infiltration after etokimab compared with placebo upon skin challenge with house dust mite, reactivity to which has been implicated in the pathogenesis of AD. We showed that etokimab also inhibited neutrophil migration to skin interstitial fluid in vitro. Besides direct effects on neutrophil migration, etokimab revealed additional unexpected CXCR1-dependent effects on IL-8–induced neutrophil migration. These human in vivo findings confirm an IL-33 upstream role in modulating skin inflammatory cascades and define the therapeutic potential for IL-33 inhibition in human diseases, including AD.

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Differential attenuation of β2 integrin–dependent and –independent neutrophil migration by Ly6G ligation

Blood Advances ◽

10.1182/bloodadvances.2018026732 ◽

2019 ◽

Vol 3 (3) ◽

pp. 256-267 ◽

Cited By ~ 2

Author(s):

Pierre Cunin ◽

Pui Y. Lee ◽

Edy Kim ◽

Angela B. Schmider ◽

Nathalie Cloutier ◽

...

Keyword(s):

Interleukin 1 ◽

Neutrophil Migration ◽

Surface Protein ◽

Joint Inflammation ◽

Selective Inhibition ◽

Β2 Integrin ◽

Β2 Integrins ◽

Neutrophil Surface

Abstract Antibody ligation of the murine neutrophil surface protein Ly6G disrupts neutrophil migration in some contexts but not others. We tested whether this variability reflected divergent dependence of neutrophil migration on β2 integrins, adhesion molecules that interact with Ly6G at the neutrophil surface. In integrin-dependent murine arthritis, Ly6G ligation attenuated joint inflammation, even though mice lacking Ly6G altogether developed arthritis normally. By contrast, Ly6G ligation had no impact on integrin-independent neutrophil migration into inflamed lung. In peritoneum, the role of β2 integrins varied with stimulus, proving dispensable for neutrophil entry in Escherichia coli peritonitis but contributory in interleukin 1 (IL-1)–mediated sterile peritonitis. Correspondingly, Ly6G ligation attenuated only IL-1 peritonitis, disrupting the molecular association between integrins and Ly6G and inducing cell-intrinsic blockade restricted to integrin-dependent migration. Consistent with this observation, Ly6G ligation impaired integrin-mediated postadhesion strengthening for neutrophils arresting on activated cremaster endothelium in vivo. Together, these findings identify selective inhibition of integrin-mediated neutrophil emigration through Ly6G ligation, highlighting the marked site and stimulus specificity of β2 integrin dependence in neutrophil migration.

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Differential Consequences of Bmp9 Deletion on Sinusoidal Endothelial Cell Differentiation and Liver Fibrosis in 129/Ola and C57BL/6 Mice

Cells ◽

10.3390/cells8091079 ◽

2019 ◽

Vol 8 (9) ◽

pp. 1079 ◽

Cited By ~ 6

Author(s):

Agnès Desroches-Castan ◽

Emmanuelle Tillet ◽

Nicolas Ricard ◽

Marie Ouarné ◽

Christine Mallet ◽

...

Keyword(s):

Endothelial Cell ◽

Liver Fibrosis ◽

Premature Mortality ◽

Suitable Model ◽

Mrna Levels ◽

Liver Sinusoidal Endothelial Cells ◽

Differentiation Markers ◽

Liver Endothelial Cell ◽

Endothelial Cell Differentiation

The aim of the present work was to address the role of BMP9 in different genetic backgrounds (C57BL/6, BALB/c, and 129/Ola) of mice deleted for Bmp9. We found that Bmp9 deletion led to premature mortality only in the 129/Ola strain. We have previously shown that Bmp9 deletion led to liver sinusoidal endothelial cells (LSEC) capillarization and liver fibrosis in the 129/Ola background. Here, we showed that this is not the case in the C57BL/6 background. Analysis of LSEC from Wild-type (WT) versus Bmp9-KO mice in the C57BL/6 background showed no difference in LSEC fenestration and in the expression of differentiation markers. Comparison of the mRNA expression of LSEC differentiation markers between WT C57BL/6 and 129/Ola mice showed a significant decrease in Stabilin2, Plvap, and CD209b, suggesting a more capillary-like phenotype in WT C57BL/6 LSECs. C57BL/6 mice also had lower BMP9 circulating concentrations and hepatic Vegfr2 mRNA levels, compared to the 129/Ola mice. Taken together, our observations support a role for BMP9 in liver endothelial cell fenestration and prevention of fibrosis that is dependent on genetic background. It also suggests that 129/Ola mice are a more suitable model than C57BL/6 for the study of liver fibrosis subsequent to LSEC capillarization.

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Regulation of dendritic cell migration and adaptive immune response by leukotriene B4 receptors: a role for LTB4 in up-regulation of CCR7 expression and function

Blood ◽

10.1182/blood-2006-02-003665 ◽

2006 ◽

Vol 109 (2) ◽

pp. 626-631 ◽

Cited By ~ 83

Author(s):

Annalisa Del Prete ◽

Wen-Hai Shao ◽

Stefania Mitola ◽

Giuseppe Santoro ◽

Silvano Sozzani ◽

...

Keyword(s):

Gene Array ◽

Adaptive Immune Response ◽

Leukotriene B4 ◽

Contact Hypersensitivity ◽

Mrna Levels ◽

Peripheral Tissues ◽

Ccr7 Expression ◽

Adaptive Immune

Abstract Trafficking of dendritic cells (DCs) to peripheral tissues and to secondary lymphoid organs depends on chemokines and lipid mediators. Here, we show that bone marrow–derived DCs (BM-DCs) express functional leukotriene B4 (LTB4) receptors as observed in dose-dependent chemotaxis and calcium mobilization responses. LTB4, at low concentrations, promoted the migration of immature and mature DCs to CCL19 and CCL21, which was associated with a rapid (30-minute) increase of CCR7 expression at the membrane level. At longer incubation times (6 hours), gene array analysis revealed a promoting role of LTB4, showing a significant increase of CCR7 and CCL19 mRNA levels. BM-DCs cultured from BLT1−/− or BLT1/2−/− mice showed a normal phenotype, but in vivo BLT1/2−/−DCs showed dramatic decrease in migration to the draining lymph nodes relative to wild-type (WT) DCs. Consistent with these observations, BLT1/2−/− mice showed a reduced response in a model of 2,4-dinitro-fluorobenzene (DNFB)–induced contact hypersensitivity. Adoptive transfer of 2,4-dinitrobenzene sulfonic acid (DNBS)–pulsed DCs directly implicated the defect in DC migration to lymph node with the defect in contact hypersensitivity. These results provide strong evidence for a role of LTB4 in regulating DC migration and the induction of adaptive immune responses.

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Discovery of methylfarnesoate as the annelid brain hormone reveals an ancient role of sesquiterpenoids in reproduction

eLife ◽

10.7554/elife.17126 ◽

2016 ◽

Vol 5 ◽

Cited By ~ 9

Author(s):

Sven Schenk ◽

Christian Krauditsch ◽

Peter Frühauf ◽

Christopher Gerner ◽

Florian Raible

Keyword(s):

Egg Production ◽

Target Species ◽

Hormone Activity ◽

Transcript Levels ◽

Reproductive Maturation ◽

Central Energy ◽

Molecular Signals ◽

Bristle Worms

Animals require molecular signals to determine when to divert resources from somatic functions to reproduction. This decision is vital in animals that reproduce in an all-or-nothing mode, such as bristle worms: females committed to reproduction spend roughly half their body mass for yolk and egg production; following mass spawning, the parents die. An enigmatic brain hormone activity suppresses reproduction. We now identify this hormone as the sesquiterpenoid methylfarnesoate. Methylfarnesoate suppresses transcript levels of the yolk precursor Vitellogenin both in cell culture and in vivo, directly inhibiting a central energy–costly step of reproductive maturation. We reveal that contrary to common assumptions, sesquiterpenoids are ancient animal hormones present in marine and terrestrial lophotrochozoans. In turn, insecticides targeting this pathway suppress vitellogenesis in cultured worm cells. These findings challenge current views of animal hormone evolution, and indicate that non-target species and marine ecosystems are susceptible to commonly used insect larvicides.

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Role of Untranslated Regions of the Hemagglutinin-Neuraminidase Gene in Replication and Pathogenicity of Newcastle Disease Virus

Journal of Virology ◽

10.1128/jvi.00188-09 ◽

2009 ◽

Vol 83 (11) ◽

pp. 5943-5946 ◽

Cited By ~ 22

Author(s):

Yongqi Yan ◽

Subrat N. Rout ◽

Shin-Hee Kim ◽

Siba K. Samal

Keyword(s):

Newcastle Disease Virus ◽

Disease Virus ◽

Newcastle Disease ◽

Untranslated Regions ◽

Mrna Levels ◽

Recombinant Viruses ◽

Virus Particles

ABSTRACT To determine the role of untranslated regions (UTRs) in replication and pathogenesis of Newcastle disease virus (NDV), we generated recombinant viruses with deletions in 5′ and 3′ UTRs of the HN mRNA. Deletion of any HN UTR did not noticeably affect in vitro replication of these viruses. However, complete deletion of the 5′ UTR of the HN gene decreased the HN mRNA levels and HN protein contents in virus particles, resulting in attenuation of the virus in chickens. This indicates that the 5′ UTR of HN mRNA plays an important role in replication and pathogenicity of NDV in vivo.

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