scholarly journals Tracking the virus-like particles of Macrobrachium rosenbergii nodavirus in insect cells

PeerJ ◽  
2017 ◽  
Vol 5 ◽  
pp. e2947 ◽  
Author(s):  
Ummi Fairuz Hanapi ◽  
Chean Yeah Yong ◽  
Zee Hong Goh ◽  
Noorjahan Banu Alitheen ◽  
Swee Keong Yeap ◽  
...  
Keyword(s):  
Spodoptera Frugiperda ◽  
Live Cell Imaging ◽  
Insect Cells ◽  
Macrobrachium Rosenbergii ◽  
Effective Vaccine ◽  
Sf9 Cells ◽  
Nuclear Localisation Signal ◽  
Virus Like Particles ◽  
Internalisation Process ◽  
Insight Into

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.

scholarly journals Immune Responses in Mice Vaccinated with Virus-Like Particles of Western Equine Encephalitis Virus from an Insect Cell-Based Expression System

2021 ◽  
Author(s):  
Jinzhu Ma ◽  
Hualei Wang ◽  
Xuexing Zheng ◽  
Hongxia Wu Wu ◽  
Songtao Yang ◽  
...  
Keyword(s):  
Insect Cells ◽  
High Titer ◽  
Expression System ◽  
Encephalitis Virus ◽  
Effective Vaccine ◽  
Virus Like Particles ◽  
Equine Encephalitis Virus ◽  
Important Challenge ◽  
Western Equine Encephalitis Virus ◽  
Western Equine Encephalitis

Western equine encephalitis virus (WEEV) can cause lethal encephalitis in humans and equines and represents a serious public health threat in many countries. Therefore, development of efficient vaccines against WEEV remains an important challenge in the field of disease control. This study described for the first time successful production of WEEV virus-like particles (VLPs) in insect cells using recombinant baculoviruses. This well-established expression system is very suitable for production of WEEV VLPs. The immune experiment herein in mice showed that the VLPs formulated with 206-adjuvant were responsible for the stronger-VLP-specific cellular immune response, and were able to induce the secretion of IL-2, IL-4, IFN-γ and production of high titer antibodies that can effectively neutralize the WEEV pseudoviruses. The WEEV VLPs from insect cells could provide a new, safe, non-replicating and effective vaccine candidate against WEEV infections.

scholarly journals Co-expression of the capsid proteins of Cowpea mosaic virus in insect cells leads to the formation of virus-like particles

2000 ◽  
Vol 81 (12) ◽  
pp. 3093-3097 ◽  
Author(s):  
Michael Shanks ◽  
George P. Lomonossoff
Keyword(s):  
Mosaic Virus ◽  
Spodoptera Frugiperda ◽  
Insect Cells ◽  
Gradient Centrifugation ◽  
Coat Proteins ◽  
Cowpea Mosaic Virus ◽  
Virion Assembly ◽  
Virus Like Particles ◽  
Assembly Pathway ◽  
The Individual

The regions of RNA-2 of Cowpea mosaic virus (CPMV) that encode the Large (L) and Small (S) coat proteins were expressed either individually or together in Spodoptera frugiperda (sf21) cells using baculovirus vectors. Co-expression of the two coat proteins from separate promoters in the same construct resulted in the formation of virus-like particles whose morphology closely resembled that of native CPMV virions. No such particles were formed when the individual L and S proteins were expressed. Sucrose gradient centrifugation of the virus-like particles showed that they had the sedimentation characteristics of empty (protein-only) shells. The results confirm that the 60 kDa L–S fusion is not an obligate intermediate in the virion assembly pathway and indicate that expression of the coat proteins in insect cells will provide a fruitful route for the study of CPMV morphogenesis.

Critical Insight into the Attributes of Emerging Novel Coronavirus (COVID-19) in India and Across the World

Coronaviruses ◽  
2020 ◽  
Vol 1 (1) ◽  
pp. 49-56
Author(s):  
Gaurav M. Doshi ◽  
Hemen S. Ved ◽  
Ami P. Thakkar
Keyword(s):  
Review Article ◽  
World Health ◽  
Effective Vaccine ◽  
Human Contact ◽  
The World ◽  
Critical Insight ◽  
Novel Coronavirus ◽  
Health Organization ◽  
Insight Into ◽  
Old Drugs

The World Health Organization (WHO) has recently announced the spread of novel coronavirus (nCoV) globally and has declared it a pandemic. The probable source of transmission of the virus, which is from animal to human and human to human contact, has been established. As per the statistics reported by the WHO on 11th April 2020, data has shown that more than sixteen lakh confirmed cases have been identified globally. The reported cases related to nCoV in India have been rising substantially. The review article discusses the characteristics of nCoV in detail with the probability of potentially effective old drugs that may inhibit the virus. The research may further emphasize and draw the attention of the world towards the development of an effective vaccine as well as alternative therapies. Moreover, the article will help to bridge the gap between the new researchers since it’s the current thrust area of research.

scholarly journals Cell adaptive response to extracellular matrix density is controlled by ICAP-1–dependent β1-integrin affinity

2008 ◽  
Vol 180 (2) ◽  
pp. 427-441 ◽  
Author(s):  
Angélique Millon-Frémillon ◽  
Daniel Bouvard ◽  
Alexei Grichine ◽  
Sandra Manet-Dupé ◽  
Marc R. Block ◽  
...  
Keyword(s):  
Conformational Changes ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Β1 Integrin ◽  
Embryonic Fibroblasts ◽  
Matrix Surface ◽  
Extracellular Microenvironment ◽  
And Migration ◽  
Integrated Response ◽  
Insight Into

Cell migration is an integrated process requiring the continuous coordinated assembly and disassembly of adhesion structures. How cells orchestrate adhesion turnover is only partially understood. We provide evidence for a novel mechanistic insight into focal adhesion (FA) dynamics by demonstrating that integrin cytoplasmic domain–associated protein 1 (ICAP-1) slows down FA assembly. Live cell imaging, which was performed in both Icap-1–deficient mouse embryonic fibroblasts and cells expressing active β1 integrin, shows that the integrin high affinity state favored by talin is antagonistically controlled by ICAP-1. This affinity switch results in modulation in the speed of FA assembly and, consequently, of cell spreading and migration. Unexpectedly, the ICAP-1–dependent decrease in integrin affinity allows cell sensing of matrix surface density, suggesting that integrin conformational changes are important in mechanotransduction. Our results clarify the function of ICAP-1 in cell adhesion and highlight the central role it plays in the cell's integrated response to the extracellular microenvironment.

Diagnostic aptitude of West Nile virus-like particles expressed in insect cells

2018 ◽  
Vol 91 (3) ◽  
pp. 233-238 ◽  
Author(s):  
Belén Rebollo ◽  
Javier Sarraseca ◽  
Mª José Rodríguez ◽  
Antonio Sanz ◽  
Miguel Ángel Jiménez-Clavero ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Insect Cells ◽  
West Nile ◽  
Virus Like Particles
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