Inhibition of the HCV Core Protein on the Immune Response to HBV Surface Antigen and on HBV Gene Expression and Replication In Vivo

Background/Objectives: In the Middle East, people consume camel milk regularly as it is believed to improve immunity against diseases and decrease the risk for cancer. Recently, it was noted that most of the beneficial effects of milk come from their nanoparticles, especially exosomes. Herein, we evaluated the anticancer potential of camel milk and its exosomes on MCF7 breast cancer cells (in vitro and in vivo) and investigated the possible underlying molecular mechanism of action. Methods/Results: Administration of camel milk (orally) and its exosomes (orally and by local injection) decreased breast tumor progression as evident by ( a) higher apoptosis (indicated by higher DNA fragmentation, caspase-3 activity, Bax gene expression, and lower Bcl2 gene expression), ( b) remarkable inhibition of oxidative stress (decrease in MDA levels and iNOS gene expression); ( c) induction of antioxidant status (increased activities of SOD, CAT, and GPX), ( d) notable reduction in expression of inflammation-( IL1b, NFκB), angiogenesis-( VEGF) and metastasis-( MMP9, ICAM1) related genes; and ( e) higher immune response (high number of CD+4, CD+8, NK1.1 T cells in spleen). Conclusions: Overall, administration of camel milk–derived exosomes showed better anticancer effect, but less immune response, than treatment by camel milk. Moreover, local injection of exosomes led to better improvement than oral administration. These findings suggest that camel milk and its exosomes have anticancer effect possibly through induction of apoptosis and inhibition of oxidative stress, inflammation, angiogenesis and metastasis in the tumor microenvironment. Thus, camel milk and its exosomes could be used as an anticancer agent for cancer treatment.

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In Vivo Modulation of Angiogenesis and Immune Response on a Collagen Matrix via Extracorporeal Shockwaves

International Journal of Molecular Sciences ◽

10.3390/ijms21207574 ◽

2020 ◽

Vol 21 (20) ◽

pp. 7574 ◽

Cited By ~ 2

Author(s):

Diana Heimes ◽

Nadine Wiesmann ◽

Jonas Eckrich ◽

Juergen Brieger ◽

Stefan Mattyasovszky ◽

...

Keyword(s):

Gene Expression ◽

Immune Response ◽

Chorioallantoic Membrane ◽

Extracorporeal Shock Wave Therapy ◽

Collagen Matrix ◽

Cam Assay ◽

Immunological Responses ◽

Tissue Integration ◽

Extracorporeal Shock Wave

The effective management of tissue integration and immunological responses to transplants decisively co-determines the success of soft and hard tissue reconstruction. The aim of this in vivo study was to evaluate the eligibility of extracorporeal shock wave therapy (ESWT) with respect to its ability to modulate angiogenesis and immune response to a collagen matrix (CM) for tissue engineering in the chorioallantoic membrane (CAM) assay, which is performed with fertilized chicken eggs. CM were placed on the CAM on embryonic development day (EDD) 7; at EDD-10, ESWT was conducted at 0.12 mJ/mm2 with 500 impulses each. One and four days later, angiogenesis represented by vascularized area, vessel density, and vessel junctions as well as HIF-1α and VEGF gene expression were evaluated. Furthermore, immune response (iNOS2, MMP-9, and MMP-13 via qPCR) was assessed and compared between ESWT- and non-ESWT-groups. At EDD-14, the vascularized area (+115% vs. +26%) and the increase in vessel junctions (+751% vs. +363%) were significantly higher in the ESWT-group. ESWT significantly increased MMP-9 gene expression at EDD-11 and significantly decreased MMP-13 gene expression at EDD-14 as compared to the controls. Using the CAM assay, an enhanced angiogenesis and neovascularization in CM after ESWT were observed. Furthermore, ESWT could reduce the inflammatory activity after a latency of four days.

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Changes in gene expression profile by HCV core protein in cultured liver cells: analysis by DNA array assay

Hepatology Research ◽

10.1016/s1386-6346(03)00017-2 ◽

2003 ◽

Vol 25 (4) ◽

pp. 396-408 ◽

Cited By ~ 7

Author(s):

K Ohkawa

Keyword(s):

Gene Expression ◽

Gene Expression Profile ◽

Expression Profile ◽

Core Protein ◽

Liver Cells ◽

Dna Array ◽

Hcv Core Protein ◽

Cultured Liver Cells

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Effect of IFN-γ on the immune response in vivo and on gene expression in vitro

Nature ◽

10.1038/307381a0 ◽

1984 ◽

Vol 307 (5949) ◽

pp. 381-382 ◽

Cited By ~ 103

Author(s):

Masataka Nakamura ◽

Tim Manser ◽

Gregory D. N. Pearson ◽

Michael J. Daley ◽

Malcolm L. Gefter

Keyword(s):

Gene Expression ◽

Immune Response ◽

Ifn Γ

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CD40 Ligand-Dependent Activation of Cytotoxic T Lymphocytes by Adeno-Associated Virus Vectors In Vivo: Role of Immature Dendritic Cells

Journal of Virology ◽

10.1128/jvi.74.17.8003-8010.2000 ◽

2000 ◽

Vol 74 (17) ◽

pp. 8003-8010 ◽

Cited By ~ 81

Author(s):

Yi Zhang ◽

Narendra Chirmule ◽

Guang-ping Gao ◽

James Wilson

Keyword(s):

Gene Expression ◽

Immune Response ◽

Dendritic Cells ◽

Cd40 Ligand ◽

Broad Host Range ◽

Mouse Bone Marrow ◽

Adeno Associated Virus ◽

Ctl Response ◽

Cell Immunity

ABSTRACT Recombinant adeno-associated virus type 2 (rAAV) is being explored as a vector for gene therapy because of its broad host range, good safety profile, and persistent transgene expression in vivo. However, accumulating evidence indicates that administration of AAV vector may initiate a detectable cellular and humoral immune response to its transduced neo-antigen in vivo. To elucidate the cellular basis of the AAV-mediated immune response, C57BL/6 mouse bone marrow-derived immature and mature dendritic cells (DCs) were infected with AAV encoding β-galactosidase (AAV-lacZ) and adoptively transferred into mice that had received an intramuscular injection of AAV-lacZ 10 days earlier. Unexpectedly, C57BL/6 mice but not CD40 ligand-deficient (CD40L−/−) mice adoptively transferred with AAV-lacZ-infected immature DCs developed a β-galactosidase-specific cytotoxic T-lymphocyte (CTL) response that markedly diminished AAV-lacZ-transduced gene expression in muscle fibers. In contrast, adoptive transfer of AAV-lacZ-infected mature DCs failed to elicit a similar CTL response in vivo. Our findings indicate, for the first time, that immature DCs may be able to elicit a CD40L-dependent T-cell immunity to markedly diminish AAV-lacZ transduced gene expression in vivo when a sufficient number of DCs capturing rAAV vector and/or its transduced gene products is recruited.

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E6AP Ubiquitin Ligase Mediates Ubiquitylation and Degradation of Hepatitis C Virus Core Protein

Journal of Virology ◽

10.1128/jvi.01684-06 ◽

2006 ◽

Vol 81 (3) ◽

pp. 1174-1185 ◽

Cited By ~ 86

Author(s):

Masayuki Shirakura ◽

Kyoko Murakami ◽

Tohru Ichimura ◽

Ryosuke Suzuki ◽

Tetsu Shimoji ◽

...

Keyword(s):

Ubiquitin Ligase ◽

Core Protein ◽

Protein Levels ◽

The Core ◽

Hcv Core Protein ◽

Ubiquitin Proteasome ◽

Proteasome Pathway ◽

Viral Nucleocapsid

ABSTRACT Hepatitis C virus (HCV) core protein is a major component of viral nucleocapsid and a multifunctional protein involved in viral pathogenesis and hepatocarcinogenesis. We previously showed that the HCV core protein is degraded through the ubiquitin-proteasome pathway. However, the molecular machinery for core ubiquitylation is unknown. Using tandem affinity purification, we identified the ubiquitin ligase E6AP as an HCV core-binding protein. E6AP was found to bind to the core protein in vitro and in vivo and promote its degradation in hepatic and nonhepatic cells. Knockdown of endogenous E6AP by RNA interference increased the HCV core protein level. In vitro and in vivo ubiquitylation assays showed that E6AP promotes ubiquitylation of the core protein. Exogenous expression of E6AP decreased intracellular core protein levels and supernatant HCV infectivity titers in the HCV JFH1-infected Huh-7 cells. Furthermore, knockdown of endogenous E6AP by RNA interference increased intracellular core protein levels and supernatant HCV infectivity titers in the HCV JFH1-infected cells. Taken together, our results provide evidence that E6AP mediates ubiquitylation and degradation of HCV core protein. We propose that the E6AP-mediated ubiquitin-proteasome pathway may affect the production of HCV particles through controlling the amounts of viral nucleocapsid protein.

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Hepatitis C Virus Core Protein Interacts with 14-3-3 Protein and Activates the Kinase Raf-1

Journal of Virology ◽

10.1128/jvi.74.4.1736-1741.2000 ◽

2000 ◽

Vol 74 (4) ◽

pp. 1736-1741 ◽

Cited By ~ 112

Author(s):

Hiroshi Aoki ◽

Junpei Hayashi ◽

Mitsuhiko Moriyama ◽

Yasuyuki Arakawa ◽

Okio Hino

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Growth Regulation ◽

Core Protein ◽

Dependent Manner ◽

Kinase Activation ◽

Virus Core ◽

Hcv Core Protein

ABSTRACT Persistent hepatitis C virus (HCV) infection is a major cause of chronic liver dysfunction in humans and is epidemiologically closely associated with the development of human hepatocellular carcinoma. Among HCV components, core protein has been reported to be implicated in cell growth regulation both in vitro and in vivo, although mechanisms explaining those effects are still unclear. In the present study, we identified that members of the 14-3-3 protein family associate with HCV core protein. 14-3-3 protein bound to HCV core protein in a phosphoserine-dependent manner. Introduction of HCV core protein caused a substantial increase in Raf-1 kinase activity in HepG2 cells and in a yeast genetic assay. Furthermore, the HCV core–14-3-3 interaction was essential for Raf-1 kinase activation by HCV core protein. These results suggest that HCV core protein may represent a novel type of Raf-1 kinase-activating protein through its interaction with 14-3-3 protein and may contribute to hepatocyte growth regulation.

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Nanosized Particles Assembled by a Recombinant Virus Protein Are Able to Encapsulate Negatively Charged Molecules and Structured RNA

Polymers ◽

10.3390/polym13060858 ◽

2021 ◽

Vol 13 (6) ◽

pp. 858

Author(s):

Hemalatha Mani ◽

Yi-Cheng Chen ◽

Yen-Kai Chen ◽

Wei-Lin Liu ◽

Shih-Yen Lo ◽

...

Keyword(s):

Self Assembly ◽

Rna Binding ◽

Core Protein ◽

Average Diameter ◽

Virus Protein ◽

Force Microscopy ◽

Atomic Force ◽

Hcv Core Protein

RNA-based molecules have recently become hot candidates to be developed into therapeutic agents. However, successful applications of RNA-based therapeutics might require suitable carriers to protect the RNA from enzymatic degradation by ubiquitous RNases in vivo. Because of their better biocompatibility and biodegradability, protein-based nanoparticles are considered to be alternatives to their synthetic polymer-based counterparts for drug delivery. Hepatitis C virus (HCV) core protein has been suggested to be able to self-assemble into nucleocapsid-like particles in vitro. In this study, the genomic RNA-binding domain of HCV core protein consisting of 116 amino acids (p116) was overexpressed with E. coli for investigation. The recombinant p116 was able to assemble into particles with an average diameter of approximately 27 nm, as visualized by electron microscopy and atomic force microscopy. Measurements with fluorescence spectroscopy, flow cytometry, and fluorescence quenching indicated that the p116-assembled nanoparticles were able to encapsulate small anionic molecules and structured RNA. This study demonstrates methods that exploit the self-assembly nature of a virus-derived protein for nanoparticle production. This study also suggests that the virus-derived protein-assembled particles could possibly be developed into potential carriers for anionic molecular drugs and structured RNA-based therapeutics.

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