Testosterone promotes Coxsackievirus B3 pathogenesis in an oral inoculation mouse model

Enteroviruses initiate infection in the gastrointestinal tract, and sex is often a biological variable that impacts infection. The role of sex hormones on enterovirus pathogenesis, however, is unclear. Previous data indicate that sex hormones can influence intestinal replication of Coxsackievirus B3 (CVB3), an enterovirus in the Picornavirus family. To determine if testosterone promotes CVB3 infection, male mice were castrated and provided placebo or testosterone-filled capsules. We found that testosterone-treated mice shed significantly more CVB3 in the feces and succumbed to CVB3-induced disease at a higher rate than castrated mice given a placebo. Treatment of male mice with an androgen receptor antagonist, flutamide, protected male mice from CVB3-induced lethality, further confirming the role of testosterone in viral pathogenesis. We also observed higher viral loads in peripheral tissues of testosterone-treated mice and an increase in the cytokine and chemokine response. Finally, we found that testosterone treatment in female mice increased fecal CVB3 shedding but had no impact on viral lethality. Overall, these data indicate that testosterone and androgen receptor signaling can promote CVB3 replication in the intestine and enhance CVB3 lethality in a sex-dependent manner.

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Androgen Receptor-Dependent and Independent Atheroprotection by Testosterone in Male Mice

Endocrinology ◽

10.1210/en.2010-0663 ◽

2010 ◽

Vol 151 (11) ◽

pp. 5428-5437 ◽

Cited By ~ 58

Author(s):

Johan Bourghardt ◽

Anna S. K. Wilhelmson ◽

Camilla Alexanderson ◽

Karel De Gendt ◽

Guido Verhoeven ◽

...

Keyword(s):

Androgen Receptor ◽

Apolipoprotein E ◽

High Fat Diet ◽

Atherosclerotic Lesion ◽

Male Mice ◽

Wild Type ◽

High Fat ◽

Lesion Area ◽

Major Pathway

The atheroprotective effect of testosterone is thought to require aromatization of testosterone to estradiol, but no study has adequately addressed the role of the androgen receptor (AR), the major pathway for the physiological effects of testosterone. We used AR knockout (ARKO) mice on apolipoprotein E-deficient background to study the role of the AR in testosterone atheroprotection in male mice. Because ARKO mice are testosterone deficient, we sham operated or orchiectomized (Orx) the mice before puberty, and Orx mice were supplemented with placebo or a physiological testosterone dose. From 8 to 16 wk of age, the mice consumed a high-fat diet. In the aortic root, ARKO mice showed increased atherosclerotic lesion area (+80%, P < 0.05). Compared with placebo, testosterone reduced lesion area both in Orx wild-type (WT) mice (by 50%, P < 0.001) and ARKO mice (by 24%, P < 0.05). However, lesion area was larger in testosterone-supplemented ARKO compared with testosterone-supplemented WT mice (+57%, P < 0.05). In WT mice, testosterone reduced the presence of a necrotic core in the plaque (80% among placebo-treated vs. 12% among testosterone-treated mice; P < 0.05), whereas there was no significant effect in ARKO mice (P = 0.20). In conclusion, ARKO mice on apolipoprotein E-deficient background display accelerated atherosclerosis. Testosterone treatment reduced atherosclerosis in both WT and ARKO mice. However, the effect on lesion area and complexity was more pronounced in WT than in ARKO mice, and lesion area was larger in ARKO mice even after testosterone supplementation. These results are consistent with an AR-dependent as well as an AR-independent component of testosterone atheroprotection in male mice.

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Protective Role of Naturally Occurring Interleukin-17A-Producing γδ T Cells in the Lung at the Early Stage of Systemic Candidiasis in Mice

Infection and Immunity ◽

10.1128/iai.05799-11 ◽

2011 ◽

Vol 79 (11) ◽

pp. 4503-4510 ◽

Cited By ~ 55

Author(s):

Takashi Dejima ◽

Kensuke Shibata ◽

Hisakata Yamada ◽

Hiromitsu Hara ◽

Yoichiro Iwakura ◽

...

Keyword(s):

T Cells ◽

Early Stage ◽

Γδ T Cells ◽

Protective Role ◽

Dependent Manner ◽

Peripheral Tissues ◽

Content Type ◽

Naturally Occurring ◽

Interleukin 17A

ABSTRACTInterleukin-17A (IL-17A)-producing γδ T cells differentiate in the fetal thymus and reside in the peripheral tissues, such as the lungs of naïve adult mice. We show here that naturally occurring γδ T cells play a protective role in the lung at a very early stage after systemic infection withCandida albicans.Selective depletion of neutrophils byin vivoadministration of anti-Ly6G monoclonal antibody (MAb) impaired fungal clearance more prominently in the lung than in the kidney 24 h after intravenous infection withC. albicans.Rapid and transient production of IL-23 was detected in the lung at 12 h, preceding IL-17A production and the influx of neutrophils, which reached a peak at 24 h after infection. IL-17A knockout (KO) mice showed reduced infiltration of neutrophils concurrently with impaired fungal clearance in the lung after infection. The major source of IL-17A was the γδ T cell population in the lung, and Cδ KO mice showed little IL-17A production and reduced neutrophil infiltration after infection. Early IL-23 production in a TLR2/MyD88-dependent manner and IL-23-triggered tyrosine kinase 2 (Tyk2) signaling were essential for IL-17A production by γδ T cells. Thus, our study demonstrated a novel role of naturally occurring IL-17A-producing γδ T cells in the first line of host defense againstC. albicansinfection.

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CD11b is involved in coxsackievirus B3-induced viral myocarditis in mice by inducing Th17 cells

Open Life Sciences ◽

10.1515/biol-2020-0085 ◽

2020 ◽

Vol 15 (1) ◽

pp. 1024-1032

Author(s):

Heng Wei ◽

Chong-Kai Lin ◽

Sheng-Jian Lu ◽

Yu-Xin Wen ◽

Shuai Yuan ◽

...

Keyword(s):

Immune Responses ◽

Th17 Cells ◽

Cytotoxic Effect ◽

Viral Myocarditis ◽

Coxsackievirus B3 ◽

Rt Pcr ◽

Cardiac Damage ◽

Cvb3 Infection ◽

Life Threatening

AbstractViral myocarditis (VMC) caused by coxsackievirus B3 (CVB3) infection is a life-threatening disease. The cardiac damage during VMC is not mainly due to the direct cytotoxic effect of the virus on cardiomyocytes but mostly involves the induction of immune responses. Integrin CD11b plays an important role in immune response, for instance, in the induction of Th17 cells. However, the role of CD11b in the pathogenesis of VMC remains largely unknown. In the present study, a mouse model of VMC was established by CVB3 infection and CD11b was knocked down in the VMC mice by transfection with siRNA-CD11b. The expression of CD11b and IL-17 in heart tissues, frequency of Th17 cells in spleen tissues and serum IL-17 levels were measured using quantitative RT-PCR, Western blot, immunohistochemistry, flow cytometry and ELISA. Results showed that CVB3 infection caused the pathological changes in heart tissues with the increases in the following indexes: expression of CD11b and IL-17 in heart tissues, frequency of Th17 cells in spleen tissues and serum IL-17 levels. The expression of CD11b was positively correlated with IL-17 expression in heart tissues. Depletion of CD11b attenuated the damage caused by CVB3 and decreased the frequency of Th17 cells in spleen tissues as well as in IL-17, IL-23 and STAT3 expression in heart tissues. In summary, our findings reveal that disruption of CD11b function reduced CVB3-induced myocarditis, suggesting that CD11b may be a novel therapeutic target for VMC.

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Role of nitric oxide-based immunotherapy in augmenting prostate cancer progression by targeting androgen receptor heterogeneity.

Journal of Clinical Oncology ◽

10.1200/jco.2020.38.15_suppl.e17537 ◽

2020 ◽

Vol 38 (15_suppl) ◽

pp. e17537-e17537

Author(s):

Himanshu Arora ◽

Anastasia Vedenko ◽

Derek J. Van Booven ◽

Manish Narasimman

Keyword(s):

Prostate Cancer ◽

Nitric Oxide ◽

Cell Proliferation ◽

Androgen Receptor ◽

Significant Proportion ◽

Dependent Manner ◽

Independent State ◽

M1 Macrophage ◽

Androgen Independent

e17537 Background: A significant proportion of men with Prostate Cancer (PCa) develop castration resistant prostate cancer (CRPC) and do not respond to hormonal agents that decrease androgens. In trying to understand the causes of androgen resistance that develop in CRPC, it is considered most relevant to study the role of Androgen receptor (AR) in the development and progression of PCa from androgen dependent to androgen independent state. Recent studies have highlighted the significance of tumor microenvironment (TME) in regulation of PCa progression in addition to AR. A key molecule in the regulation of TME interactions is nitric oxide (NO). We have shown in our recent study, the critical association of NO with the TME in CRPC. However, the effects of NO to modulate the progression of PCa to CRPC with respect to AR still remains largely unexplored. Methods: 22RV1, LNCaP, LNCaPAPIPC(cells expressing no AR), and LNCaPshAR/pATK (cells expressing low AR), cells were used for the study. Cell proliferation was first assessed by MTT assay. The castrated SCID mice were grafted with 22RV1 cells and were treated with GSNO at the dosage of 10mg/kg/day IP. After treatment, animals were humanely sacrificing. Tumor RNA and proteins were analysed for markers that are important for PCa progression using qPCR, western blot and cytokine antibody array. Animal experiments were carried out in compliance with the IACUC of University of Miami. GraphPad Prism (GraphPad Software) was used for statistical analysis. Results: In addition to reducing the tumor burden, the expression of anti-inflammatory (M2) macrophages (CD206 and Arginase1) is decreased and that of the pro-inflammatory (M1) macrophage (iNOS) is increased in mice which received increased NO levels. Furthermore, to study the effects of NO on progression of PCa from androgen dependent to androgen independent stage, we characterized the LNCAP cell models with differential extent of AR knockdown (LNCaP, LNCaPshAR/pATK and LNCaPAPIPC) for the effects of increased NO levels. Results showed that NO had significant impact on cell proliferation on androgen dependent PCa cells however the effects were negligible in cells expressing low or no AR, suggesting that effects of NO on PCa cell proliferation are AR dependent. Conclusions: Our results suggest that during PCa progression, NO suppresses TAMs to target the TME in an AR dependent manner. Further studies are undergoing to establish the impacts of NO in PCa progression.

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Autophagosome Supports Coxsackievirus B3 Replication in Host Cells

Journal of Virology ◽

10.1128/jvi.00641-08 ◽

2008 ◽

Vol 82 (18) ◽

pp. 9143-9153 ◽

Cited By ~ 238

Author(s):

Jerry Wong ◽

Jingchun Zhang ◽

Xiaoning Si ◽

Guang Gao ◽

Ivy Mao ◽

...

Keyword(s):

Viral Replication ◽

Protein Degradation ◽

Viral Myocarditis ◽

Membrane Vesicles ◽

Coxsackievirus B3 ◽

Host Cells ◽

Small Interfering Rnas ◽

Autophagosome Formation ◽

Cvb3 Infection

ABSTRACT Recent studies suggest a possible takeover of host antimicrobial autophagy machinery by positive-stranded RNA viruses to facilitate their own replication. In the present study, we investigated the role of autophagy in coxsackievirus replication. Coxsackievirus B3 (CVB3), a picornavirus associated with viral myocarditis, causes pronounced intracellular membrane reorganization after infection. We demonstrate that CVB3 infection induces an increased number of double-membrane vesicles, accompanied by an increase of the LC3-II/LC3-I ratio and an accumulation of punctate GFP-LC3-expressing cells, two hallmarks of cellular autophagosome formation. However, protein expression analysis of p62, a marker for autophagy-mediated protein degradation, showed no apparent changes after CVB3 infection. These results suggest that CVB3 infection triggers autophagosome formation without promoting protein degradation by the lysosome. We further examined the role of the autophagosome in CVB3 replication. We demonstrated that inhibition of autophagosome formation by 3-methyladenine or small interfering RNAs targeting the genes critical for autophagosome formation (ATG7, Beclin-1, and VPS34 genes) significantly reduced viral replication. Conversely, induction of autophagy by rapamycin or nutrient deprivation resulted in increased viral replication. Finally, we examined the role of autophagosome-lysosome fusion in viral replication. We showed that blockage of the fusion by gene silencing of the lysosomal protein LAMP2 significantly promoted viral replication. Taken together, our results suggest that the host's autophagy machinery is activated during CVB3 infection to enhance the efficiency of viral replication.

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Protein Kinase B/Akt Regulates Coxsackievirus B3 Replication through a Mechanism Which Is Not Caspase Dependent

Journal of Virology ◽

10.1128/jvi.78.8.4289-4298.2004 ◽

2004 ◽

Vol 78 (8) ◽

pp. 4289-4298 ◽

Cited By ~ 81

Author(s):

Mitra Esfandiarei ◽

Honglin Luo ◽

Bobby Yanagawa ◽

Agripina Suarez ◽

Darya Dabiri ◽

...

Keyword(s):

Protein Kinase ◽

Protein Expression ◽

Hela Cells ◽

Protein Kinase B ◽

Coxsackievirus B3 ◽

Dominant Negative ◽

Viral Rna ◽

Dominant Negative Mutant ◽

Cvb3 Infection

ABSTRACT The role of signaling pathways including the mitogen-activated protein kinases (MAPKs) and phosphatidylinositol 3-kinase (PI3K) during viral infection has gained much recent attention. Our laboratory reported on an important regulatory role for extracellular signal-regulated kinases (ERK1/2), subfamily members of the MAPKs, during coxsackievirus B3 (CVB3) infection. However, the role of the PI3K pathway in CVB3 infection has not been well characterized. CVB3 is the most common known viral infectant of heart muscle that directly injures and kills infected cardiac myocytes during the myocarditic process. In the present study, we investigated the role of protein kinase B (PKB) (also known as Akt), a general downstream mediator of survival signals through the PI3K cascade, in regulating CVB3 replication and virus-induced apoptosis in a well-established HeLa cell model. We have demonstrated that CVB3 infection leads to phosphorylation of PKB/Akt on both Ser-473 and Thr-308 residues through a PI3K-dependent mechanism. Transfection of HeLa cells with a dominant negative mutant of Akt1 or pretreatment of wild-type HeLa cells with the specific PI3K inhibitor LY294002 significantly suppresses viral RNA expression, as reflected in diminished viral capsid protein expression and viral release. Dominant negative Akt1 and LY294002 also increase apoptosis in infected cells, which can be reversed by addition of the general caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (zVAD.fmk). Interestingly, blocking of apoptosis by zVAD.fmk does not reverse the viral RNA translation blockade, indicating that the inhibitory effect of dominant negative Akt1 on viral protein expression is not caspase dependent. In addition, we showed that the attachment of virus to its receptor-coreceptor complex is not sufficient for PKB/Akt activation and that postentry viral replication is required for Akt phosphorylation. Taken together, these data illustrate a new and imperative role for Akt in CVB3 infection in HeLa cells and show that the PI3K/Akt signaling is beneficial to CVB3 replication.

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Effect of Hydroalcoholic Extract of Satchys Lavandulifolia on Pentylenetetrazole-Induced Seizures in Male Mice: The Role of Gabaergic and Opioidergic Systems

Basic and Clinical Neuroscience Journal ◽

10.32598/bcn.2021.2299.1 ◽

2021 ◽

Author(s):

Hamid Behzad Nia ◽

◽

Amin Naseri ◽

Mohammadreza Emamhadi ◽

Shervin Ghadarjani ◽

...

Keyword(s):

Normal Saline ◽

Saline Group ◽

Anticonvulsant Effect ◽

Dependent Manner ◽

Male Mice ◽

Hydroalcoholic Extract ◽

Pretreated Group ◽

Pre Treatment ◽

The Impact

Introduction: Epilepsy is one of the most common neurological disorders. Though there are effective medications available for treatment of epilepsy, the use of most drugs is associated with many side effects and drug interactions. Stachys lavandulifolia (SL) used in Iranian traditional medicine show anti-anxiety and sedative actions. The objective of the current study was to evaluate the anticonvulsant effect of hydroalcoholic extract of SL on the pentylenetetrazole (PTZ)-induced seizure in male mice and the role of benzodiazepine and opioid receptors. Methods: This study was conducted on 100 male mice randomly categorized into 10 groups: Normal Saline, Diazepam groups (0.025 and 0.1 mg/kg), SL extract groups (50, 100 and 200 mg/kg), Diazepam 0.025 mg/kg + SL extract 50mg/kg and three groups that pre-treated with NS, Flumazenil or Naloxone, 5 min before injection of 200 mg/kg extract. After 30 min, PTZ (80 mg/kg) was injected to animals and seizure indices were evaluated. Results: The SL extract attenuated the PTZ-induced seizures in a dose dependent manner and pre-treatment with flumazenil reversed this effect of SL extract but pre-treatment with naloxone could not reverse this effect, because seizure indices on naloxone pretreated group was still lower than normal saline group. Combination of ineffective dose of diazepam and SL extract decrease PTZ-induced seizures. Discussion: The results of our study showed the anticonvulsant properties of hydroalcoholic extract of SL. These effects might be due to the impact of the components of this extract on the central benzodiazepine system.

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The role of nesfatin-1 in metabolism regulation: an overview / Rola nesfatyny-1 w regulacji metabolizmu: artykuł przeglądowy

Annals of Animal Science ◽

10.2478/aoas-2013-0002 ◽

2013 ◽

Vol 13 (2) ◽

pp. 195-205

Author(s):

Michał Bulc ◽

Sławomir Gonkowski ◽

Jarosław Całka

Keyword(s):

Body Weight Gain ◽

Wide Distribution ◽

Dark Phase ◽

Dependent Manner ◽

Peripheral Tissues ◽

Functional Studies ◽

Metabolism Regulation ◽

Dose Dependent ◽

The Brain

Abstract The hypothalamus synthesizes molecules involved in the regulation of feeding behaviour. Nesfatin- 1 is a recently discovered substance expressed in both the brain and peripheral tissues and exerts a strong anorectic action. Nesfatin-1-immunoreactive cell bodies are distributed in arcuate (ARC), paraventricular (PVN) and supraoptic (SON) nuclei, where the peptide has been found to be co-expressed with pro-opiomelanocortin (POMC) and cocaine- and amphetamine-regulated transcript (CART), oxytocin (OX) and vasopressin (VP). More detailed studies have shown a wide distribution of nesfatin-1-positive neurons in several brain areas, such as the forebrain, hindbrain, brainstem and spinal cord. Moreover, nesfatin-1 has been also expressed in peripheral tissues, colocalizing with ghrelin in the gastric mucosa and insulin in β-cells of the endocrine pancreas and adipose tissue. Functional studies have revealed that exogenous nesfatin-1 administered into the brain ventricles, subcutaneously or intraperitoneally, was able to decrease both food intake in the dark phase as well as body weight gain in a dose-dependent manner. In addition, recent findings suggest the involvement of nesfatin-1 in the control of insulin secretion as well as immune and stress-related responses. However, since there is still a deficiency of data concerning the nesfatin-1 receptor, the possible implementation of nesfatin-1 analogs during human metabolic disorders requires further study.

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