The Experts below are selected from a list of 126 Experts worldwide ranked by ideXlab platform
Rhonda Cardin - One of the best experts on this subject based on the ideXlab platform.
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The Coxsackie Virus and Adenovirus Receptor (Car) is a Required Host Factor for Recovirus Infection : a Putative Enteric Calicivirus Receptor.
Journal of Virology, 2019Co-Authors: Tibor Farkas, Kui Yang, Jacques Le Pendu, Joel Baines, Rhonda CardinAbstract:Human norovirus (HuNoV) is a leading cause of acute gastroenteritis in both developed and developing countries. Studies of HuNoV host Cell interactions are limited by the lack of a simple, robust Cell culture system. Due to their diverse HuNoV-like biological features, including histo-blood group antigen (HBGA) binding, rhesus enteric caliciviruses (ReCVs) are viable surrogate models for HuNoVs. In addition, several ReCVs strains can be propagated to high titers in standard non-human Primate Cell lines while causing lytic infection and Cell death. To identify the ReCV entry receptor, we performed CRISPR/Cas9 library screening in Vero Cells which identified the coxsackie virus and adenovirus receptor (CAR) as a candidate ReCV entry receptor. We showed that siRNA, anti-hCAR Mab RmcB treatment, and recombinant hCAR ectodomain blocked ReCV replication in LLC-MK2 Cells. CRISPR/Cas9 targeted knockout of CAR in LLC-MK2 and Vero Cells made these Cell lines resistant to ReCV infection and susceptibility to infection could be restored by transient expression of CAR. CHO Cells do not express CAR or HBGAs and are resistant to ReCV infection. Recombinant CHO Cells stably expressing hCAR or the type B HBGA alone did not support ReCV infection. However, CHO Cells expressing both hCAR and the type B HBGA were susceptible to ReCV infection. In summary, we have demonstrated that CAR is required for ReCV infection and most likely is a functional ReCV receptor, but HBGAs are also necessary for infection.IMPORTANCE Because of the lack of a simple and robust human norovirus (HuNoV) Cell culture system surrogate caliciviruses still represent valuable research tools for norovirus research. Due to their remarkable biological similarities to HuNoVs, including the utilization of HBGAs as putative attachment receptors, we used rhesus enteric caliciviruses (ReCVs) to study enteric calicivirus host Cell interactions. Using CRISPR/Cas9 library screening and functional assays we identified and validated the coxsackie virus and adenovirus receptor (CAR) as a functional proteinaceous receptor for ReCVs. Our work demonstrated that CAR and HBGAs are both necessary to convert a non-susceptible Cell line susceptible to ReCV infection. Follow up studies to evaluate the involvement of CAR in HuNoV infections are ongoing.
Tibor Farkas - One of the best experts on this subject based on the ideXlab platform.
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The Coxsackie Virus and Adenovirus Receptor (Car) is a Required Host Factor for Recovirus Infection : a Putative Enteric Calicivirus Receptor.
Journal of Virology, 2019Co-Authors: Tibor Farkas, Kui Yang, Jacques Le Pendu, Joel Baines, Rhonda CardinAbstract:Human norovirus (HuNoV) is a leading cause of acute gastroenteritis in both developed and developing countries. Studies of HuNoV host Cell interactions are limited by the lack of a simple, robust Cell culture system. Due to their diverse HuNoV-like biological features, including histo-blood group antigen (HBGA) binding, rhesus enteric caliciviruses (ReCVs) are viable surrogate models for HuNoVs. In addition, several ReCVs strains can be propagated to high titers in standard non-human Primate Cell lines while causing lytic infection and Cell death. To identify the ReCV entry receptor, we performed CRISPR/Cas9 library screening in Vero Cells which identified the coxsackie virus and adenovirus receptor (CAR) as a candidate ReCV entry receptor. We showed that siRNA, anti-hCAR Mab RmcB treatment, and recombinant hCAR ectodomain blocked ReCV replication in LLC-MK2 Cells. CRISPR/Cas9 targeted knockout of CAR in LLC-MK2 and Vero Cells made these Cell lines resistant to ReCV infection and susceptibility to infection could be restored by transient expression of CAR. CHO Cells do not express CAR or HBGAs and are resistant to ReCV infection. Recombinant CHO Cells stably expressing hCAR or the type B HBGA alone did not support ReCV infection. However, CHO Cells expressing both hCAR and the type B HBGA were susceptible to ReCV infection. In summary, we have demonstrated that CAR is required for ReCV infection and most likely is a functional ReCV receptor, but HBGAs are also necessary for infection.IMPORTANCE Because of the lack of a simple and robust human norovirus (HuNoV) Cell culture system surrogate caliciviruses still represent valuable research tools for norovirus research. Due to their remarkable biological similarities to HuNoVs, including the utilization of HBGAs as putative attachment receptors, we used rhesus enteric caliciviruses (ReCVs) to study enteric calicivirus host Cell interactions. Using CRISPR/Cas9 library screening and functional assays we identified and validated the coxsackie virus and adenovirus receptor (CAR) as a functional proteinaceous receptor for ReCVs. Our work demonstrated that CAR and HBGAs are both necessary to convert a non-susceptible Cell line susceptible to ReCV infection. Follow up studies to evaluate the involvement of CAR in HuNoV infections are ongoing.
G Kaplan - One of the best experts on this subject based on the ideXlab platform.
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Susceptibility of nonPrimate Cell lines to hepatitis A virus infection.
Journal of virology, 1994Co-Authors: A Dotzauer, S M Feinstone, G KaplanAbstract:Abstract Hepatitis A virus (HAV) has been adapted to grow in Primate Cell cultures. We investigated replication of HAV in nonPrimate Cells by inoculating 20 Cell lines from different species with the tissue culture-adapted HM175 strain. Slot blot hybridization and immunofluorescence analysis revealed that HAV replicated in GPE, SP 1K, and IB-RS-2 D10 Cells of guinea pig, dolphin, and pig origin, respectively. Studies in IB-RS-2 D10 Cells were discontinued because cultures were contaminated with classical swine fever virus. A growth curve showed that HAV grew poorly in GPE Cells and intermediately in SP 1K Cells compared with growth in FRhK-4 Cells. Therefore, the Cell surface receptor(s) and other host factor(s) required for HAV replication are present in nonPrimate as well as Primate Cells.
Greg J Towers - One of the best experts on this subject based on the ideXlab platform.
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restriction of multiple divergent retroviruses by lv1 and ref1
The EMBO Journal, 2003Co-Authors: Theodora Hatziioannou, Simone Cowan, Stephen P Goff, Paul D Bieniasz, Greg J TowersAbstract:The mouse gene Fv1 encodes a saturable restriction factor that selectively blocks infection by N-tropic or B-tropic murine leukemia virus (MLV) strains. Despite the absence of an Fv1 gene, a similar activity is present in humans that blocks N-MLV infection (Ref1). Moreover, some non-human Primate Cell lines express a potentially related inhibitor of HIV-1 and/or SIVmac infection (Lv1). Here, we examine the spectrum of retrovirus-restricting activities expressed by human and African green monkey Cell lines. Human Cells restrict N-MLV and equine infectious anemia virus (EIAV), but not HIV-1, HIV-2, SIVmac or SIVagm, whilst AGM Cells restrict N-MLV, EIAV, HIV-1, HIV-2 and SIVmac. Remarkably, in each example examined, restriction of infection by a given retrovirus can be abrogated at least partially by saturation with another retrovirus, provided that it is also restricted but regardless of whether it is closely related. These data suggest that restriction factors in human and non-human Primate Cells are able to recognize and block infection by multiple, widely divergent retroviruses and that the factors themselves may be related.
Joseph S. M. Peiris - One of the best experts on this subject based on the ideXlab platform.
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Recurrent mutations associated with isolation and passage of SARS coronavirus in Cells from non-human Primates.
Journal of medical virology, 2005Co-Authors: Leo L.m. Poon, Cynthia S. W. Leung, Kwok H. Chan, Kwok-yung Yuen, Yi Guan, Joseph S. M. PeirisAbstract:Four clinical isolates of SARS coronavirus were serially passaged in two Primate Cell lines (FRhK4 and Vero E6). Viral genetic sequences encoding for structural proteins and open reading frames 6–8 were determined in the original clinical specimen, the initial virus isolate (passage 0) and at passages 5, 10, and 15. After 15 passages, a total of 15 different mutations were identified and 12 of them were non-synonymous mutations. Seven of these mutations were recurrent mutation and all located at the spike, membrane, and Orf 8a protein encoding sequences. Mutations in the membrane protein and a deletion in ORF 6–8 were already observed in passage 0, suggesting these amino acid substitutions are important in the adaptation of the virus isolate in Primate Cell culture. A mutation in the spike gene (residue 24079) appeared to be unique to adaptation in FRhK4 Cells. It is important to be aware of Cell culture associated mutations when interpreting data on molecular evolution of SARS coronavirus. J. Med. Virol. 76:435–440, 2005. © 2005 Wiley-Liss, Inc.