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Generation of recombinant rabies virus ERA strain applied to virus tracking in cell infection
ZHAO Dan-dan, SHUAI Lei, GE Jin-ying, WANG Jin-liang, WEN Zhi-yuan, LIU Ren-qiang, WANG Chong, WANG Xi-jun, BU Zhi-gao
2019, 18 (10): 2361-2368.   DOI: 10.1016/S2095-3119(19)62717-6
Abstract156)      PDF in ScienceDirect      
The mechanism of rabies virus (RABV) infection still needs to be further characterized.  RABV particle with self-fluorescent is a powerful viral model to visualize the viral infection process in cells.  Herein, based on a reverse genetic system of the Evelyn-Rokitnicki-Abelseth (rERA) strain, we generated a recombinant RABV rERA-N/mCherry strain that stably expresses an additional ERA nucleoprotein that fuses with the red fluorescent protein mCherry (N/mCherry).  The rERA-N/mCherry strain retained growth property similar to the parent strain rERA in vitro.  The N/mCherry expression showed genetic stability during passage into mouse neuroblastoma (NA) cells and did not change the virulence of the vector.  The rERA-N/mCherry strain was then utilized as a visual viral model to study the RABV-cell binding and internalization.  We directly observed the red self-fluorescence of rERA-N/mCherry particles binding to the cell surface, and further co-localizing with clathrin in the early stage of infection in NA cells by fluorescence microscopy.  Our results showed that the rERA-N/mCherry strain uses clathrin-dependent endocytosis to enter cells, which is consistent with the well-known mechanism of RABV invasion.  The recombinant RABV rERA-N/mCherry thus appears to have the potential to be an effective viral model to further explore the fundamental molecular mechanism of rabies neuropathogenesis.
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Newcastle disease virus-based MERS-CoV candidate vaccine elicits high-level and lasting neutralizing antibodies in Bactrian camels
Liu Ren-qiang, Ge Jin-ying, Wang Jin-ling, Shao Yu, Zhang Hui-lei, Wang Jin-liang, Wen Zhi-yuan, Bu Zhi-gao
2017, 16 (10): 2264-2273.   DOI: 10.1016/S2095-3119(17)61660-5
Abstract578)      PDF in ScienceDirect      
Middle East respiratory syndrome coronavirus (MERS-CoV), a member of the Coronaviridae family, is the causative pathogen for MERS that is characterized by high fever, pneumonia, acute respiratory distress syndrome (ARDS), as well as extrapulmonary manifestations.  Currently, there are no approved treatment regimens or vaccines for MERS.  Here, we generated recombinant nonvirulent Newcastle disease virus (NDV) LaSota strain expressing MERS-CoV S protein (designated as rLa-MERS-S), and evaluated its immunogenicity in mice and Bactrian camels.  The results revealed that rLa-MERS-S showed similar growth properties to those of LaSota in embryonated chicken eggs, while animal immunization studies showed that rLa-MERS-S induced MERS-CoV neutralizing antibodies in mice and camels.  Our findings suggest that recombinant rLa-MERS-S may be a potential MERS-CoV veterinary vaccine candidate for camels and other animals affected by MERS.
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