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Masataka Tsuge, Nobuhiko Hiraga, Yizhou Zhang, Misa Yamashita, Ojiro Sato, Naoya Oka, Kanma Shiraishi, Yu Izaki, Grace Naswa Makokha, Takuro Uchida, Mio Kurihara, Motonobu Nomura, Ken Tsushima, Takashi Nakahara, Eisuke Murakami, Hiromi Abe-Chayama, Tomokazu Kawaoka, Daiki Miki, Michio Imamura, Yoshiiku Kawakami, Hiroshi Aikata, Hidenori Ochi, C Nelson Hayes, Takashi Fujita, Kazuaki Chayama
The events in the immune response to hepatitis B virus (HBV) remain unclear. We analyzed the direct influence of HBV on gene expression in human hepatocytes under immunodeficient conditions using a human hepatocyte chimeric mouse model. HBV-infected or non-infected chimeric mouse livers were collected, and gene expression profiles were compared. Since IL-8 was the most significantly up-regulated gene at 8 weeks after HBV infection, we focused on IL-8 and found that HBx and the large HBs (L-HBs) protein induce transcription of IL-8 via endoplasmic reticulum stress...
September 18, 2018: Virology
Liang Cui, Madina Mahesutihan, Weinan Zheng, Lijun Meng, Wenhui Fan, Jing Li, Xin Ye, Wenjun Liu, Lei Sun
Cell division cycle 25 B (CDC25B) is a member of the CDC25 phosphatase family. It can dephosphorylate cyclin-dependent kinases and regulate the cell division cycle. Moreover, siRNA knockdown of CDC25B impairs influenza A virus (IAV) replication. Here, to further understand the regulatory mechanism of CDC25B for IAV replication, a CDC25B-knockout (KO) 293T cell line was constructed using CRISPR/Cas9. The present data indicated that the replication of IAV was decreased in CDC25B-KO cells. Additionally, CDC25B deficiency damaged viral polymerase activity, nucleoprotein (NP) self-oligomerization, and NP nuclear export...
September 18, 2018: Virology
Lei Li, Thomas J DeLiberto, Mary L Killian, Mia K Torchetti, Xiu-Feng Wan
In March 2017, a novel highly pathogenic avian influenza A(H7N9) virus was detected at two commercial broiler breeder facilities in Tennessee, United States. In this study, a wild bird low pathogenic avian influenza A virus, A/blue-winged teal/Wyoming/AH0099021/2016(H7N9), was shown to be the probable precursor of the novel H7N9 virus; this low pathogenic virus has eight possible progenitor genes sharing > 99% sequence identity with the novel H7N9 virus. Phylogeographic analyses showed that viral gene constellations that formed and circulated among dabbling ducks contributed to the emergence of the novel H7N9 virus...
September 17, 2018: Virology
Yurong Qu, Yuan Zhan, Shen Yang, Shanhui Ren, Xusheng Qiu, Zaib Ur Rehamn, Lei Tan, Yingjie Sun, Chunchun Meng, Cuiping Song, Shengqing Yu, Chan Ding
High mobility group box 1 (HMGB1) is a key member of the "danger associated molecular patterns" (DAMPs) which plays important roles in systemic inflammation and has a pathogenic role in infectious diseases like viral or bacterial infections. Newcastle disease virus (NDV) infection is proved to cause intense inflammatory responses that result in excessive cellular apoptosis and tissue damage, but the function of HMGB1 in NDV-induced cytokine storm has not been elucidated. Here, we report that HMGB1 is a significant inflammation factor in NDV infection...
September 11, 2018: Virology
Francisco Javier Del Toro, Eva Mencía, Emmanuel Aguilar, Francisco Tenllado, Tomas Canto
Native and amino acid (aa) substitution mutants of HCPro from potato virus Y (PVY) were transiently expressed in Nicotiana benthamiana leaves. Properties of those HCPro variants with regard to silencing suppression activities, mediation of viral transmission by aphids, and subcellular localization dynamics, were determined. One mutant failed to suppress silencing in agropatch assays, but could efficiently mediate the transmission by aphids of purified virions. This mutant also retained the ability to translocate to microtubules (MTs) in stressed cells...
September 10, 2018: Virology
Yongli Xiao, Jacqueline M Nolting, Zong-Mei Sheng, Tyler Bristol, Li Qi, Andrew S Bowman, Jeffery K Taubenberger
Influenza virus infections in humans and animals are major public health concerns. In the current study, a set of universal influenza enrichment probes was developed to increase the sensitivity of sequence-based virus detection and characterization for all influenza viruses. This universal influenza enrichment probe set contains 46,953 120nt RNA biotin-labeled probes designed based on all available influenza viral sequences and it can be used to enrich for influenza sequences without prior knowledge of type or subtype...
September 10, 2018: Virology
Amornrat O'Brien, Robert C Mettelman, Aaron Volk, Nicole M André, Gary R Whittaker, Susan C Baker
Investigating type I feline coronaviruses (FCoVs) in tissue culture is critical for understanding the basic virology, pathogenesis, and virus-host interactome of these important veterinary pathogens. This has been a perennial challenge as type I FCoV strains do not easily adapt to cell culture. Here we characterize replication kinetics and plaque formation of a model type I strain FIPV Black in Fcwf-4 cells established at Cornell University (Fcwf-4 CU). We determined that maximum virus titers (>107 pfu/mL) were recoverable from infected Fcwf-4 CU cell-free supernatant at 20 h post-infection...
September 8, 2018: Virology
Ki-Hye Kim, Young-Man Kwon, Young-Tae Lee, Hye Suk Hwang, Min-Chul Kim, Eun-Ju Ko, Bao-Zhong Wang, Fu-Shi Quan, Sang-Moo Kang
Current licensed adjuvants including aluminum hydroxide (alum) bias immune responses toward T helper type 2 (Th2) immune responses. We tested whether virus-like particles presenting flagellin (Flag VLP) exhibit adjuvant effects on eliciting Th1 type immune responses and improving the efficacy of poor immunogenic tandem repeat M2e (M2e5x) protein vaccine against influenza virus. Co-immunization of mice with Flag VLP and M2e5x protein vaccine induced significantly higher levels of IgG2a isotype (Th1) antibodies in sera and mucosal sites, effector CD4+ T cells secreting IFN-γ and granzyme B, and more effective lung viral clearance and protection compared to alum adjuvant...
September 7, 2018: Virology
Khulud Bukhari, Geraldine Mulley, Anastasia A Gulyaeva, Lanying Zhao, Guocheng Shu, Jianping Jiang, Benjamin W Neuman
Transcriptomics has the potential to discover new RNA virus genomes by sequencing total intracellular RNA pools. In this study, we have searched publicly available transcriptomes for sequences similar to viruses of the Nidovirales order. We report two potential nidovirus genomes, a highly divergent 35.9 kb likely complete genome from the California sea hare Aplysia californica, which we assign to a nidovirus named Aplysia abyssovirus 1 (AAbV), and a coronavirus-like 22.3 kb partial genome from the ornamented pygmy frog Microhyla fissipes, which we assign to a nidovirus named Microhyla alphaletovirus 1 (MLeV)...
September 7, 2018: Virology
M Mura, C Ruffié, E Billon-Denis, C Combredet, J N Tournier, F Tangy
Measles virus has been successfully attenuated on chicken embryo cells to obtain a highly efficient and safe live attenuated vaccine, administered thus far to billions of children. Measles virus attenuation has long been described to involve a modification of cellular tropism with the use of human CD46 ubiquitous receptor. Nevertheless, the use of this receptor in vivo is not obvious. In this study we use four different mouse models to decipher the respective part of hCD46 receptor and type-I interferon response in measles host restriction...
September 7, 2018: Virology
Yingfen Lei, Kazuyo Takeda, Li Yu
Flavivirus RNA replication starts at 3'-end, where it folds into a highly conserved stem-loop structure. We attempted to identify the viral non-structural proteins (NSPs) that might specifically interact with the 3'-stemloop (3'SL) through a genetic approach. WNV/DENV2 chimeric recombinants that contain Dengue2 (DENV2) gene(s) in West Nile virus (WNV) backbone were tested for replication competence. Three of seven recombinant viruses, containing the DENV2 NS1, NS2A, or NS4B gene and terminated with a mutated 3'SL (MutC 3'SL), were viable...
September 5, 2018: Virology
Dinesh Babu Paudel, Basudev Ghoshal, Sushma Jossey, Marta Ludman, Karoly Fatyol, Hélène Sanfaçon
ARGONAUTEs (notably AGO1 and AGO2) are effectors of plant antiviral RNA silencing. AGO1 was shown to be required for the temperature-dependent symptom recovery of Nicotiana benthamiana plants infected with tomato ringspot virus (isolate ToRSV-Rasp1) at 27 °C. In this study, we show that symptom recovery from isolate ToRSV-GYV shares similar hallmarks of antiviral RNA silencing but occurs at a wider range of temperatures (21-27 °C). At 21 °C, an early spike in AGO2 mRNAs accumulation was observed in plants infected with either ToRSV-Rasp1 or ToRSV-GYV but the AGO2 protein was only consistently detected in ToRSV-GYV infected plants...
September 5, 2018: Virology
Xiaoyun Liu, Yi Liu, Qiuyan Zhang, Bo Zhang, Han Xia, Zhiming Yuan
Subgenomic flavivirus RNA (sfRNA) is essential for flavivirus-induced cytopathicity. sfRNA-deficient mutant dengue virus (DENV) M8 harboring a deleted domain II in the 3' untranslated region (UTR) has severely decreased cytopathicity and replication efficiency. Here, recovered viruses acquired from M8 by serial passages replicated similarly to the wild type (WT) but produced two types of sfRNA, whereas only one was detected in WT infected cells, resulting in mildly decreased cytopathicity. Sequencing and structure prediction showed that recovered viruses acquired a novel additional partial domain I sequence duplication and formed analogous RNA secondary structures...
August 30, 2018: Virology
S M Bywaters, S A Brendle, J Biryukov, J W Wang, J Walston, J Milici, R B Roden, C Meyers, N D Christensen
The major capsid protein of HPV, L1, assembles into pentamers that form a T = 7 icosahedral particle, but the location of the co-assembled minor capsid protein, L2, remains controversial. Several researchers have developed useful monoclonal antibodies targeting L2, but most react with linear epitopes toward the N-terminus. As a means to better define the virus capsid and better assess the localization and exposure of L2 epitopes in the context of assembled HPV, we have developed a panel of 30 monoclonal antibodies (mAbs) which target the N-terminus of L2 amino acids 11-200, previously defined as a broadly protective immunogen...
August 28, 2018: Virology
Sonia Zicari, Anush Arakelyan, Rogers Alberto Ñahui Palomino, Wendy Fitzgerald, Christophe Vanpouille, Anna Lebedeva, Alain Schmitt, Morgane Bomsel, William Britt, Leonid Margolis
Extracellular vesicles (EVs) released by virus-infected cells typically incorporate host and viral components inside the vesicles (cargo molecules). Here, we investigated if human cytomegalovirus (HCMV) proteins are incorporated in EV outer membrane released by HCMV-infected cells. We separated EVs from HCMV using an iodixanol step-gradient and found that the separated vesicles carried EV markers such as the tetraspanin CD63 and Rab27A. Flow analysis of individual EVs demonstrated that on average, 15 ± 3...
August 27, 2018: Virology
Keisuke Ohta, Yusuke Matsumoto, Natsuko Yumine, Machiko Nishio
We previously demonstrated that human parainfluenza virus type 2 (hPIV-2) induces RhoA activation, which promotes its growth. RhoA controls the equilibrium between globular and filamentous actin (F-actin). We found that F-actin formation is induced by wild type (wt) hPIV-2 infection, and that inhibition of F-actin formation by cytochalasin D decreases hPIV-2 growth. In wt RhoA-expressing cells, F-actin formation occurs and hPIV-2 growth is promoted. Overexpression of T19N RhoA, a dominant negative (DN) form of RhoA, inhibits hPIV-2-induced F-actin formation, and suppresses hPIV-2 growth...
August 27, 2018: Virology
Rui Guo, Pengcheng Shang, Celena A Carrillo, Zhi Sun, Yashavanth Shaan Lakshmanappa, Xingyu Yan, Gourapura J Renukaradhya, Jodi McGill, Crystal J Jaing, Megan C Niederwerder, Raymond R R Rowland, Ying Fang
In order to study the mechanism of PRRSV persistence, an in vitro model of persistence was developed by serially passaging PRRSV-infected MARC-145 cells 109 times. Viral persistence was detected to be associated with increased double-stranded (dsRNA) in the infected cells. In PRRSV infected pigs, reduced ratio of plus to minus strands of viral RNA was observed in lymphoid tissues from PRRSV persistent pigs at 52 days post infection. Viral dsRNA was mostly detected in the germinal center during persistent infection compared to the localization of dsRNA in the inter-follicular zones during acute infection...
August 27, 2018: Virology
Vladimir V Zarubaev, Ekaterina A Pushkina, Sophia S Borisevich, Anastasia V Galochkina, Angelica V Garshinina, Anna A Shtro, Anna A Egorova, Anastasia S Sokolova, Sergey L Khursan, Olga I Yarovaya, Nariman F Salakhutdinov
Due to the ability of influenza virus to develop drug resistance, the search for novel antivirals is an important goal of medical science and health care systems. We assessed the ability of the influenza virus to develop resistance to the hemagglutinin inhibitor camphecene and characterized laboratory-selected resistant strains. We showed by electron microscopy that camphecene decreases the number of virions fusing their envelopes with endosomal membranes. A 160-fold decrease in virus susceptibility was observed after six passages in cells...
August 27, 2018: Virology
Guochao Wei, Timo Kehl, Qiuying Bao, Axel Benner, Janet Lei, Martin Löchelt
The retroviral Gag protein, the major component of released particles, plays different roles in particle assembly, maturation or infection of new host cells. Here, we characterize the Gag chromatin binding site including the highly conserved QPQRYG motif of feline foamy virus, a member of the Spumaretrovirinae. Mutagenesis of critical residues in the chromatin binding site/QPQRYG motif almost completely abrogates viral DNA integration and reduces nuclear accumulation of Gag and viral DNA. Genome packaging, reverse transcription, particle release and uptake into new target cells are not affected...
August 23, 2018: Virology
Jessica A Belser, Taronna R Maines, Terrence M Tumpey
Reconstruction of the 1918 influenza virus has facilitated considerable advancements in our understanding of this extraordinary pandemic virus. However, the benefits of virus reconstruction are not limited to this one strain. Here, we provide an overview of laboratory studies which have evaluated the reconstructed 1918 virus, and highlight key discoveries about determinants of virulence and transmissibility associated with this virus in mammals. We further discuss recent and current pandemic threats from avian and swine reservoirs, and provide specific examples of how reconstruction of the 1918 pandemic virus has improved our ability to contextualize research employing novel and emerging strains...
August 21, 2018: Virology
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