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Porcine deltacoronavirus (PDCoV) is an emerging infectious disease of swine with zoonotic potential. Phylogenetic analysis suggests that PDCoV originated recently from a host-switching event between birds and mammals. Little is known about how PDCoV interacts with its differing hosts. Human-derived cell lines are susceptible to PDCoV infection. Herein, we compare the gene expression profiles of an established host swine cells to potential emerging host human cells after infection with PDCoV. Cell lines derived from intestinal lineages were used to reproduce the primary sites of viral infection in the host. Porcine intestinal epithelial cells (IPEC-J2) and human intestinal epithelial cells (HIEC) were infected with PDCoV. RNA-sequencing was performed on total RNA extracted from infected cells. Human cells exhibited a more pronounced response to PDCoV infection in comparison to porcine cells with more differentially expressed genes (DEGs) in human, 7486, in comparison to pig cells, 1134. On the transcriptional level, the adoptive host human cells exhibited more DEGs in response to PDCoV infection in comparison to the primary pig host cells, where different types of cytokines can control PDCoV replication and virus production. Key immune-associated DEGs and signaling pathways are shared between human and pig cells during PDCoV infection. These included genes related to the NF-kappa-B transcription factor family, the interferon (IFN) family, the protein-kinase family, and signaling pathways such as the apoptosis signaling pathway, JAK-STAT signaling pathway, inflammation/cytokine–cytokine receptor signaling pathway. MAP4K4 was unique in up-regulated DEGs in humans in the apoptosis signaling pathway. While similarities exist between human and pig cells in many pathways, our research suggests that the adaptation of PDCoV to the porcine host required the ability to down-regulate many response pathways including the interferon pathway. Our findings provide an important foundation that contributes to an understanding of the mechanisms of PDCoV infection across different hosts. To our knowledge, this is the first report of transcriptome analysis of human cells infected by PDCoV.
Diana Cruz-Pulido; Patricia A. Boley; Wilberforce Zachary Ouma; Moyasar Alhamo; Linda J. Saif; Scott P. Kenney. Comparative Transcriptome Profiling of Human and Pig Intestinal Epithelial Cells after Porcine Deltacoronavirus Infection. Viruses 2021, 13, 292 .
AMA StyleDiana Cruz-Pulido, Patricia A. Boley, Wilberforce Zachary Ouma, Moyasar Alhamo, Linda J. Saif, Scott P. Kenney. Comparative Transcriptome Profiling of Human and Pig Intestinal Epithelial Cells after Porcine Deltacoronavirus Infection. Viruses. 2021; 13 (2):292.
Chicago/Turabian StyleDiana Cruz-Pulido; Patricia A. Boley; Wilberforce Zachary Ouma; Moyasar Alhamo; Linda J. Saif; Scott P. Kenney. 2021. "Comparative Transcriptome Profiling of Human and Pig Intestinal Epithelial Cells after Porcine Deltacoronavirus Infection." Viruses 13, no. 2: 292.
Hepatitis E virus (HEV) can account for up to a 30% mortality rate in pregnant women, with highest incidences reported for genotype 1 (gt1) HEV. Reasons contributing to adverse maternal-fetal outcome during pregnancy in HEV-infected pregnant women remain elusive in part due to the lack of a robust tissue culture model for some strains. Open reading frame (ORF4) was discovered overlapping ORF1 in gt1 HEV whose protein expression is regulated via an IRES-like RNA element. To experimentally determine whether gt3 HEV contains an ORF4-like gt1, gt1 and gt3 sequence comparisons were performed between the gt1 and the homologous gt3 sequence. To assess whether ORF4 protein could enhance gt3 replication, Huh7 cell lines constitutively expressing ORF4 were created and used to assess the replication of the Kernow-C1 gt3 and sar55 gt1 HEV. Virus stocks from transfected Huh7 cells with or without ORF4 were harvested and infectivity assessed via infection of HepG2/C3A cells. We also studied the replication of gt1 HEV in the ORF4-expressing tunicamycin-treated cell line. To directly show that HEV transcripts have productively replicated in the target cells, we assessed events at the single-cell level using indirect immunofluorescence and flow cytometry. Despite not naturally encoding ORF4, replication of gt3 HEV was enhanced by the presence of gt1 ORF4 protein. These results suggest that the function of ORF4 protein from gt1 HEV is transferrable, enhancing the replication of gt3 HEV. ORF4 may be utilized to enhance replication of difficult to propagate HEV genotypes in cell culture. IMPORTANCE: HEV is a leading cause of acute viral hepatitis (AVH) around the world. The virus is a threat to pregnant women, particularly during the second and third trimester of pregnancy. The factors enhancing virulence to pregnant populations are understudied. Additionally, field strains of HEV remain difficult to culture in vitro. ORF4 was recently discovered in gt1 HEV and is purported to play a role in pregnancy related pathology and enhanced replication. We present evidence that ORF4 protein provided in trans enhances the viral replication of gt3 HEV even though it does not encode ORF4 naturally in its genome. These data will aid in the development of cell lines capable of supporting replication of non-cell culture adapted HEV field strains, allowing viral titers sufficient for studying these strains in vitro. Furthermore, development of gt1/gt3 ORF4 chimeric virus may shed light on the role that ORF4 plays during pregnancy.
Kush K. Yadav; Patricia A. Boley; Zachary Fritts; Scott P. Kenney. Ectopic Expression of Genotype 1 Hepatitis E Virus ORF4 Increases Genotype 3 HEV Viral Replication in Cell Culture. Viruses 2021, 13, 75 .
AMA StyleKush K. Yadav, Patricia A. Boley, Zachary Fritts, Scott P. Kenney. Ectopic Expression of Genotype 1 Hepatitis E Virus ORF4 Increases Genotype 3 HEV Viral Replication in Cell Culture. Viruses. 2021; 13 (1):75.
Chicago/Turabian StyleKush K. Yadav; Patricia A. Boley; Zachary Fritts; Scott P. Kenney. 2021. "Ectopic Expression of Genotype 1 Hepatitis E Virus ORF4 Increases Genotype 3 HEV Viral Replication in Cell Culture." Viruses 13, no. 1: 75.
Coronaviruses cause respiratory and gastrointestinal diseases in diverse host species. Deltacoronaviruses (DCoVs) have been identified in various songbird species and in leopard cats in China. In 2009, porcine deltacoronavirus (PDCoV) was detected in fecal samples from pigs in Asia, but its etiologic role was not identified until 2014, when it caused major diarrhea outbreaks in swine in the United States. Studies have shown that PDCoV uses a conserved region of the aminopeptidase N protein to infect cell lines derived from multiple species, including humans, pigs, and chickens. Because PDCoV is a potential zoonotic pathogen, investigations of its prevalence in humans and its contribution to human disease continue. We report experimental PDCoV infection and subsequent transmission among poultry. In PDCoV-inoculated chicks and turkey poults, we observed diarrhea, persistent viral RNA titers from cloacal and tracheal samples, PDCoV-specific serum IgY antibody responses, and antigen-positive cells from intestines.
Patricia A. Boley; Moyasar A. Alhamo; Geoffrey Lossie; Kush Kumar Yadav; Marcia Vasquez-Lee; Linda J. Saif; Scott P. Kenney. Porcine Deltacoronavirus Infection and Transmission in Poultry, United States1. Emerging Infectious Diseases 2020, 26, 255 -265.
AMA StylePatricia A. Boley, Moyasar A. Alhamo, Geoffrey Lossie, Kush Kumar Yadav, Marcia Vasquez-Lee, Linda J. Saif, Scott P. Kenney. Porcine Deltacoronavirus Infection and Transmission in Poultry, United States1. Emerging Infectious Diseases. 2020; 26 (2):255-265.
Chicago/Turabian StylePatricia A. Boley; Moyasar A. Alhamo; Geoffrey Lossie; Kush Kumar Yadav; Marcia Vasquez-Lee; Linda J. Saif; Scott P. Kenney. 2020. "Porcine Deltacoronavirus Infection and Transmission in Poultry, United States1." Emerging Infectious Diseases 26, no. 2: 255-265.
Lettuce is often implicated in human norovirus (HuNoV) foodborne outbreaks. We identified H-like histo-blood group antigens (HBGAs) on lettuce leaves as specific binding moieties for virus-like particles (VLPs) of HuNoV GII.4/HS194/2009 strain. The objective of this study was to determine whether HuNoV-lettuce binding is mediated through the virus HBGA binding sites (HBS). Toward this objective, VLPs of historical HuNoV GII.4 strains (1987, 1997, 2002, 2004 and 2006) with known natural mutations in their HBS, two newly generated VLP mutants of GII.4/HS194/2009 (D374A and G443A) and a VLP mutant (W375A) of GI.1/Norwalk/1968 along with its wild type VLPs, which displays distinct HBS, were investigated for their binding to lettuce. ELISA revealed that historical GII.4 strains binding to lettuce was dependent on their HBGAs profiles. The VLP mutants D374A and G443A lost binding to HBGAs and displayed no to minimal binding to lettuce, respectively. The VLPs of GI.1/Norwalk/1968 strain bound to lettuce through an H-like HBGA and the binding was inhibited by fucosidase digestion. Mutant W375A which was previously shown not to bind to HBGAs, displayed significantly reduced binding to lettuce. We conclude that the binding of HuNoV GII.4 and GI.1 strains to lettuce is mediated through the virus HBS.
Malak A. Esseili; Xiang Gao; Patricia Boley; Yixuan Hou; Linda J. Saif; Paul Brewer-Jensen; Lisa C. Lindesmith; Ralph S. Baric; Robert L. Atmar; Qiuhong Wang. Human Norovirus Histo-Blood Group Antigen (HBGA) Binding Sites Mediate the Virus Specific Interactions with Lettuce Carbohydrates. Viruses 2019, 11, 833 .
AMA StyleMalak A. Esseili, Xiang Gao, Patricia Boley, Yixuan Hou, Linda J. Saif, Paul Brewer-Jensen, Lisa C. Lindesmith, Ralph S. Baric, Robert L. Atmar, Qiuhong Wang. Human Norovirus Histo-Blood Group Antigen (HBGA) Binding Sites Mediate the Virus Specific Interactions with Lettuce Carbohydrates. Viruses. 2019; 11 (9):833.
Chicago/Turabian StyleMalak A. Esseili; Xiang Gao; Patricia Boley; Yixuan Hou; Linda J. Saif; Paul Brewer-Jensen; Lisa C. Lindesmith; Ralph S. Baric; Robert L. Atmar; Qiuhong Wang. 2019. "Human Norovirus Histo-Blood Group Antigen (HBGA) Binding Sites Mediate the Virus Specific Interactions with Lettuce Carbohydrates." Viruses 11, no. 9: 833.
PEDV is the most economically important porcine enteric viral pathogen and has caused immense economic losses in the pork industries in many countries. Effective and safe vaccines are desperately required but still not available. 2′- O -MTase (nsp16) is highly conserved among coronaviruses (CoVs), and the inactivation of nsp16 in live attenuated vaccines has been attempted for several betacoronaviruses. We show that inactivation of both 2′- O -MTase and the endocytosis signal of the spike protein is an approach to designing a promising live attenuated vaccine for PEDV. The in vivo passaging data also validated the stability of the KDKE 4A -SYA mutant. KDKE 4A -SYA warrants further evaluation in sows and their piglets and may be used as a platform for further optimization. Our findings further confirmed that nsp16 can be a universal target for CoV vaccine development and will aid in the development of vaccines against other emerging CoVs.
Yixuan Hou; Hanzhong Ke; Jineui Kim; Dongwan Yoo; Yunfang Su; Patricia Boley; Juliet Chepngeno; Anastasia N. Vlasova; Linda J. Saif; Qiuhong Wang. Engineering a Live Attenuated Porcine Epidemic Diarrhea Virus Vaccine Candidate via Inactivation of the Viral 2'- O -Methyltransferase and the Endocytosis Signal of the Spike Protein. Journal of Virology 2019, 93, 1 .
AMA StyleYixuan Hou, Hanzhong Ke, Jineui Kim, Dongwan Yoo, Yunfang Su, Patricia Boley, Juliet Chepngeno, Anastasia N. Vlasova, Linda J. Saif, Qiuhong Wang. Engineering a Live Attenuated Porcine Epidemic Diarrhea Virus Vaccine Candidate via Inactivation of the Viral 2'- O -Methyltransferase and the Endocytosis Signal of the Spike Protein. Journal of Virology. 2019; 93 (15):1.
Chicago/Turabian StyleYixuan Hou; Hanzhong Ke; Jineui Kim; Dongwan Yoo; Yunfang Su; Patricia Boley; Juliet Chepngeno; Anastasia N. Vlasova; Linda J. Saif; Qiuhong Wang. 2019. "Engineering a Live Attenuated Porcine Epidemic Diarrhea Virus Vaccine Candidate via Inactivation of the Viral 2'- O -Methyltransferase and the Endocytosis Signal of the Spike Protein." Journal of Virology 93, no. 15: 1.
Porcine epidemic diarrhea virus (PEDV) causes diarrhea in all ages of pigs with 50–100% mortality rates in neonatal piglets. In the United States, inactivated and subunit PEDV vaccines for pregnant sows are available, but fail to induce sufficient protection in neonatal piglets farrowed from PEDV naïve sows. A safe and efficacious live attenuated vaccine that can prime mucosal immune responses is urgently needed. In this study, we evaluated the safety and efficacy of two attenuated PEDV vaccine candidates, the emerging non-S INDEL PEDV strain PC22A at the 100th cell culture passage level - Clone no. 4 (P100C4) and at the 120th passage level (P120), in weaned pigs. Four groups of 40-day-old weaned pigs were inoculated orally with PEDV PC22A-P3 (virulent), -P100C4, -P120, and mock, respectively, and challenged with the P3 virus at 24 days post-inoculation (dpi). After inoculation, P3 caused diarrhea in all pigs with a high level of fecal viral RNA shedding. P100C4 and P120 did not cause diarrhea in pigs, although viral RNA was detected in feces of all pigs, except for one P100C4-inoculated pig. Compared with the P120 group, P3- and P100C4-inoculated pigs had higher serum PEDV-specific IgG and viral neutralizing (VN) antibody (Ab) titers at 14 dpi. After the challenge, no pigs in the P3 group but all pigs in the P100C4, P120, and mock groups had diarrhea. Compared with the P120 group, pigs in the P100C4 group had a more rapid decline in fecal PEDV RNA shedding titers, higher titers of serum PEDV-specific IgG, IgA, and VN Abs, and higher numbers of intestinal IgA Ab-secreting cells. PEDV PC22A P100C4 and P120 were fully attenuated in weaned pigs but failed to elicit protection against virulent P3 challenge. P100C4 induced higher PEDV-specific antibody responses than P120 post inoculation resulting in a greater anamnestic response post challenge. Therefore, P100C4 potentially could be tested as a priming vaccine or be further modified using reverse genetics. It also can be administered in multiple doses or be combined with inactivated or subunit vaccines and adjuvants as a PEDV vaccination regimen, whose efficacy can be tested in the future.
Chun-Ming Lin; Shristi Ghimire; Yixuan Hou; Patricia Boley; Stephanie N. Langel; Anastasia N. Vlasova; Linda J. Saif; Qiuhong Wang. Pathogenicity and immunogenicity of attenuated porcine epidemic diarrhea virus PC22A strain in conventional weaned pigs. BMC Veterinary Research 2019, 15, 26 .
AMA StyleChun-Ming Lin, Shristi Ghimire, Yixuan Hou, Patricia Boley, Stephanie N. Langel, Anastasia N. Vlasova, Linda J. Saif, Qiuhong Wang. Pathogenicity and immunogenicity of attenuated porcine epidemic diarrhea virus PC22A strain in conventional weaned pigs. BMC Veterinary Research. 2019; 15 (1):26.
Chicago/Turabian StyleChun-Ming Lin; Shristi Ghimire; Yixuan Hou; Patricia Boley; Stephanie N. Langel; Anastasia N. Vlasova; Linda J. Saif; Qiuhong Wang. 2019. "Pathogenicity and immunogenicity of attenuated porcine epidemic diarrhea virus PC22A strain in conventional weaned pigs." BMC Veterinary Research 15, no. 1: 26.
The Notch signaling pathway enables regulation and control of development, differentiation, and homeostasis through cell-cell communication. Our investigation shows that Notch signaling directly activates the Nrf2 stress adaptive response pathway through recruitment of the Notch intracellular domain (NICD) transcriptosome to a conserved Rbpjκ site in the promoter of Nrf2. Stimulation of Notch signaling through Notch ligand expression in cells and by overexpression of the NICD in RosaNICD/−::AlbCre mice in vivo induces expression of Nrf2 and its target genes. Continuous and transient NICD expression in the liver produces a Notch-dependent cytoprotective response through direct transcriptional activation of Nrf2 signaling to rescue mice from acute acetaminophen toxicity. This response can be reversed upon genetic disruption of Nrf2. Morphological studies showed that the characteristic phenotype of high-density intrahepatic bile ducts and enlarged liver in RosaNICD/−::AlbCre mice could be at least partially reversed after Nrf2 disruption. Furthermore, the liver and bile duct phenotypes could be recapitulated with constitutive activation of Nrf2 signaling in Keap1F/F::AlbCre mice. It appears that Notch-to-Nrf2 signaling is another important determinant in liver development and function and promotes cell-cell cytoprotective signaling responses.
Nobunao Wakabayashi; John Skoko; Dionysios V. Chartoumpekis; Shoko Kimura; Stephen L. Slocum; Kentaro Noda; Dushani L. Palliyaguru; Masahiro Fujimuro; Patricia A. Boley; Yugo Tanaka; Norihisa Shigemura; Shyam Biswal; Masayuki Yamamoto; Thomas Kensler; Patricia Boley. Notch-Nrf2 Axis: Regulation of Nrf2 Gene Expression and Cytoprotection by Notch Signaling. Molecular and Cellular Biology 2013, 34, 653 -663.
AMA StyleNobunao Wakabayashi, John Skoko, Dionysios V. Chartoumpekis, Shoko Kimura, Stephen L. Slocum, Kentaro Noda, Dushani L. Palliyaguru, Masahiro Fujimuro, Patricia A. Boley, Yugo Tanaka, Norihisa Shigemura, Shyam Biswal, Masayuki Yamamoto, Thomas Kensler, Patricia Boley. Notch-Nrf2 Axis: Regulation of Nrf2 Gene Expression and Cytoprotection by Notch Signaling. Molecular and Cellular Biology. 2013; 34 (4):653-663.
Chicago/Turabian StyleNobunao Wakabayashi; John Skoko; Dionysios V. Chartoumpekis; Shoko Kimura; Stephen L. Slocum; Kentaro Noda; Dushani L. Palliyaguru; Masahiro Fujimuro; Patricia A. Boley; Yugo Tanaka; Norihisa Shigemura; Shyam Biswal; Masayuki Yamamoto; Thomas Kensler; Patricia Boley. 2013. "Notch-Nrf2 Axis: Regulation of Nrf2 Gene Expression and Cytoprotection by Notch Signaling." Molecular and Cellular Biology 34, no. 4: 653-663.