Study of the interaction of Macrobrachium rosenbergii Nodavirus and its virus-like particles (MrNV-VLPs) in Sf9 insect cells: the dependence upon caveolin-mediated endocytosis and the possible existence of a capsid ligand domain

Macrobrachium rosenbergii nodavirus (MrNv) poses a major threat to the prawn industry. Currently, no effective vaccine and treatment are available to prevent the spread of MrNv. Its infection mechanism and localisation in a host cell are also not well characterised. The MrNv capsid protein (MrNvc) produced in Escherichia coli self-assembled into virus-like particles (VLPs) resembling the native virus. Thus, fluorescein labelled MrNvc VLPs were employed as a model to study the virus entry and localisation in Spodoptera frugiperda, Sf9 cells. Through fluorescence microscopy and sub-cellular fractionation, the MrNvc was shown to enter Sf9 cells, and eventually arrived at the nucleus. The presence of MrNvc within the cytoplasm and nucleus of Sf9 cells was further confirmed by the Z-stack imaging. The presence of ammonium chloride (NH4Cl), genistein, methyl-β-cyclodextrin or chlorpromazine (CPZ) inhibited the entry of MrNvc into Sf9 cells, but cytochalasin D did not inhibit this process. This suggests that the internalisation of MrNvc VLPs is facilitated by caveolae- and clathrin-mediated endocytosis. The whole internalisation process of MrNvc VLPs into a Sf9 cell was recorded with live cell imaging. We have also identified a potential nuclear localisation signal (NLS) of MrNvc through deletion mutagenesis and verified by classical-NLS mapping. Overall, this study provides an insight into the journey of MrNvc VLPs in insect cells.

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The key molecular events during Macrobrachium rosenbergii nodavirus (MrNV) infection and replication in Sf9 insect cells

Virus Research ◽

10.1016/j.virusres.2016.06.012 ◽

2016 ◽

Vol 223 ◽

pp. 1-9 ◽

Cited By ~ 12

Author(s):

Monsicha Somrit ◽

Atthaboon Watthammawut ◽

Charoonroj Chotwiwatthanakun ◽

Wattana Weerachatyanukul

Keyword(s):

Insect Cells ◽

Macrobrachium Rosenbergii ◽

Sf9 Insect Cells ◽

Molecular Events

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Peer Review #2 of "Tracking the virus-like particles of Macrobrachium rosenbergii nodavirus in insect cells (v0.1)"

10.7287/peerj.2947v0.1/reviews/2 ◽

2017 ◽

Author(s):

R Tuma

Keyword(s):

Peer Review ◽

Insect Cells ◽

Macrobrachium Rosenbergii ◽

Virus Like Particles

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Faculty Opinions recommendation of Protection against lethal challenge by Ebola virus-like particles produced in insect cells.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1158496.618669 ◽

2009 ◽

Author(s):

Grant McFadden ◽

Kim Van Vliet

Keyword(s):

Ebola Virus ◽

Insect Cells ◽

Lethal Challenge ◽

Virus Like Particles

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Production of Japanese encephalitis virus-like particles in insect cells

Bioengineered ◽

10.4161/bioe.24514 ◽

2013 ◽

Vol 4 (6) ◽

pp. 438-442 ◽

Cited By ~ 13

Author(s):

Hideki Yamaji ◽

Eiji Konishi

Keyword(s):

Japanese Encephalitis Virus ◽

Japanese Encephalitis ◽

Insect Cells ◽

Encephalitis Virus ◽

Virus Like Particles

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Expression and Immunogenicity of Two Recombinant Fusion Proteins Comprising Foot-and-Mouth Disease Virus Structural Protein VP1 and DC-SIGN-Binding Glycoproteins

BioMed Research International ◽

10.1155/2017/7658970 ◽

2017 ◽

Vol 2017 ◽

pp. 1-9 ◽

Cited By ~ 1

Author(s):

Xinsheng Liu ◽

Jianliang Lv ◽

Yuzhen Fang ◽

Peng Zhou ◽

Yanzhen Lu ◽

...

Keyword(s):

Immune Responses ◽

Disease Virus ◽

Fusion Proteins ◽

Insect Cells ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Vp1 Protein ◽

Sf9 Insect Cells ◽

Mouth Disease Virus ◽

Foot And Mouth

Improving vaccine immunogenicity by targeting antigens to dendritic cells has recently emerged as a new design strategy in vaccine development. In this study, the VP1 gene of foot-and-mouth disease virus (FMDV) serotype A was fused with the gene encoding human immunodeficiency virus (HIV) membrane glycoprotein gp120 or C2-V3 domain of hepatitis C virus (HCV) envelope glycoprotein E2, both of which are DC-SIGN-binding glycoproteins. After codon optimization, the VP1 protein and the two recombinant VP1-gp120 and VP1-E2 fusion proteins were expressed in Sf9 insect cells using the insect cell-baculovirus expression system. Western blotting showed that the VP1 protein and two recombinant VP1-gp120 and VP1-E2 fusion proteins were correctly expressed in the Sf9 insect cells and had good reactogenicity. Guinea pigs were then immunized with the purified proteins, and the resulting humoral and cellular immune responses were analyzed. The VP1-gp120 and VP1-E2 fusion proteins induced significantly higher specific anti-FMDV antibody levels than the VP1 protein and stronger cell-mediated immune responses. This study provides a new perspective for the development of novel FMDV subunit vaccines.

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Sesquiterpene Lactones from Artemisia absinthium. Biotransformation and Rearrangement of the Insect Antifeedant 3α-hydroxypelenolide

Plants ◽

10.3390/plants10050891 ◽

2021 ◽

Vol 10 (5) ◽

pp. 891

Author(s):

Braulio M. Fraga ◽

Carmen E. Díaz ◽

María Bailén ◽

Azucena González-Coloma

Keyword(s):

Cytotoxic Activity ◽

Leptinotarsa Decemlineata ◽

Spodoptera Littoralis ◽

Culture Medium ◽

Insect Cells ◽

Sesquiterpene Lactones ◽

Enzymatic System ◽

Artemisia Absinthium ◽

Sf9 Insect Cells ◽

New Compounds

Three new compounds, the sesquiterpenes absilactone and hansonlactone and the acetophenone derivative ajenjol, have been isolated from a cultivated variety of Artemisia absinthium. In addition, the major lactone isolated, 3α-hydroxypelenolide, was biotransformed by the fungus Mucor plumbeus affording the corresponding 1β, 10α-epoxide. A cadinane derivative was formed by an acid rearrangement produced in the culture medium, but not by the enzymatic system of the fungus. Furthermore, 3α-hydroxypelenolide showed strong antifeedant effects against Leptinotarsa decemlineata and cytotoxic activity to Sf9 insect cells, while the biotransformed compounds showed antifeedant postingestive effects against Spodoptera littoralis.

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