scholarly journals Human Interferon Inducible Transmembrane Protein 3 (IFITM3) Inhibits Influenza Virus A Replication and Inflammation by Interacting with ABHD16A

2020 ◽  
Author(s):  
Liang Chen ◽  
Zhu Limei ◽  
Chen Jun
Keyword(s):  
Influenza Virus ◽  
Confocal Microscopy ◽  
Influenza Viruses ◽  
Expression Level ◽  
Hek293 Cells ◽  
Human Interferon ◽  
Yeast Two Hybrid ◽  
Related Factors ◽  
Influenza Virus A ◽  
Two Hybrid

Abstract Background: Studies have shown that human interferon inducible transmembrane proteins (hIFITMs) family proteins have broad-spectrum antiviral capabilities. Preliminary studies in our laboratory have preliminarily proved that hIFITMs have the effect of inhibiting influenza viruses. In order to further study its mechanism and role in the occurrence and development of influenza A, relevant studies have relevant studies have been carried out.Methods: Fluorescence quantitative polymerase chain reaction (PCR) detection, yeast two-hybrid test and optical confocal microscopy were used to investigate the effect of hIFITM3 on influenza virus A (IVA) replication, the interaction with human abhydrolase domain containing 16A (hABHD16A) and the expression of inflammation-related factors.Results: In HEK293 cells, overexpression of hIFITM3 protein significantly inhibited the replication of IVA at 24h, 48h, and 72h; yeast two-hybrid experiment proved that hIFITM3 interacts with hABHD16A; laser confocal microscopy observations showed that hIFITM3 and hABHD16A co-localized in cell membrane area; the expression level of inflammation-related factors in cells overexpressing hIFITM3 or hABHD16A was detected by fluorescence quantitative PCR, and the results showed that the mRNA levels of interleukin (IL)-1β, IL-6, IL-10, tumor necrosis factor (TNF)-a and for cyclooxygenase 2 (COX2) were significantly increased. But when hIFITM3/hABHD16A was co-expressed, the mRNA expression levels of these cytokines were significantly reduced except COX2. When influenza virus infected cells co-expressing hIFITM3/hABHD16A, the expression level of inflammatory factors decreased compared with the control group, indicating that hIFITM3 can play an important role in regulating inflammation balance.Conclusions: This study confirmed that hIFITM3 has an effect of inhibiting IVA replication. Furthermore, it was found that hIFITM3 interacts with hABHD16A, following which it can better inhibit the replication of influenza virus and the inflammatory response caused by the disease process.

scholarly journals Interferon Inducible Transmembrane Protein 3 Inhibits Influenza Virus Replication and Inflammation by Interacting With ABHD16A

2020 ◽  
Author(s):  
Chen Liang ◽  
Limei Zhu ◽  
Jun Chen
Keyword(s):  
Influenza Virus ◽  
Confocal Microscopy ◽  
Virus Replication ◽  
Influenza Viruses ◽  
Expression Level ◽  
Hek293 Cells ◽  
Yeast Two Hybrid ◽  
Related Factors ◽  
Influenza Virus Replication ◽  
Two Hybrid

Abstract Background: Studies have shown that human interferon inducible transmembrane proteins (IFITM) family proteins have broad-spectrum antiviral capabilities. Preliminary studies in our laboratory have preliminarily proved that IFITMs have the effect of inhibiting influenza viruses. In order to further study its mechanism and role in the occurrence and development of influenza, relevant studies have been carried out.Methods: Fluorescence quantitative polymerase chain reaction (PCR) detection, yeast two-hybrid test and optical confocal microscopy were used to investigate the effect of hIFITM3 on influenza virus replication, the interaction with human abhydrolase domain containing 16A (hABHD16A) and the expression of inflammation-related factors.Results: In HEK293 cells, overexpression of hIFITM3 protein significantly inhibited the replication of influenza virus at 24h, 48h, and 72h; yeast two-hybrid experiment proved that IFITM3 interacts with ABHD16A; laser confocal microscopy observations showed that IFITM3 and ABHD16A co-localized in cell membrane area; the expression level of inflammation-related factors in cells overexpressing hIFITM3 or hABHD16A was detected by fluorescence quantitative PCR, and the results showed that the mRNA levels of interleukin (IL)-1β, IL-6, IL-10, tumor necrosis factor (TNF)-a and cyclooxygenase 2 (COX2) were significantly increased . But when IFITM3/ABHD16A was co-expressed, the mRNA expression levels of these cytokines were significantly reduced except for COX2. When influenza virus infected cells co-expressing IFITM3/ABHD16A, the expression level of inflammatory factors decreased compared with the control group, indicating that IFITM3 can play an important role in regulating inflammation balance.Conclusions: This study confirmed that hIFITM3 has an effect of inhibiting influenza virus replication. Furthermore, it was found that hIFITM3 interacts with hABHD16A, following which it can better inhibit the replication of influenza virus and the inflammatory response caused by the disease process.

scholarly journals Human Interferon Inducible Transmembrane Protein 3 (IFITM3) Inhibits Influenza Virus A Replication and Inflammation by Interacting with ABHD16A

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Liang Chen ◽  
Limei Zhu ◽  
Jun Chen
Keyword(s):  
Influenza Virus ◽  
Influenza A ◽  
Transmembrane Protein ◽  
Influenza Viruses ◽  
Expression Level ◽  
Hek293 Cells ◽  
Human Interferon ◽  
Control Group ◽  
Mrna Levels ◽  
Membrane Area

Studies have shown that human interferon inducible transmembrane protein (hIFITMs) family proteins have broad-spectrum antiviral capabilities. Preliminary studies in our laboratory have tentatively proved that hIFITMs have the effect of inhibiting influenza viruses. In order to further study its mechanism and role in the occurrence and development of influenza A, relevant studies have been carried out. Fluorescence quantitative polymerase chain reaction (PCR) detection technology was used to observe the effect of hIFITM3 on the replication of influenza A virus (IVA) and the interaction with hABHD16A. In HEK293 cells, overexpression of hIFITM3 protein significantly inhibited the replication of IVA at 24 h, 48 h, and 72 h; yeast two-hybrid experiment proved that hIFITM3 interacts with hABHD16A; laser confocal microscopy observations showed that hIFITM3 and hABHD16A colocalized in the cell membrane area; the expression level of inflammation-related factors in cells overexpressing hIFITM3 or hABHD16A was detected by fluorescence quantitative PCR, and the results showed that the mRNA levels of interleukin- (IL-) 1β, IL-6, IL-10, tumor necrosis factor- (TNF-) α, and cyclooxygenase 2 (COX2) were significantly increased. But when hIFITM3/hABHD16A was coexpressed, the mRNA expression levels of these cytokines were significantly reduced except COX2. When influenza virus infected cells coexpressing hIFITM3/hABHD16A, the expression level of inflammatory factors decreased compared with the control group, indicating that hIFITM3 can play an important role in regulating inflammation balance. This study confirmed that hIFITM3 has an effect of inhibiting IVA replication. Furthermore, it was found that hIFITM3 interacts with hABHD16A, following which it can better inhibit the replication of influenza virus and the inflammatory response caused by the disease process.

scholarly journals Detection of severe acute respiratory syndrome coronavirus 2 and influenza viruses based on CRISPR-Cas12a

2020 ◽  
pp. 153537022096379
Author(s):  
Oraphan Mayuramart ◽  
Pattaraporn Nimsamer ◽  
Somruthai Rattanaburi ◽  
Naphat Chantaravisoot ◽  
Kritsada Khongnomnan ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Severe Acute Respiratory Syndrome ◽  
Influenza A ◽  
Limit Of Detection ◽  
Cross Reactivity ◽  
Influenza Viruses ◽  
Influenza B Virus ◽  
Influenza B ◽  
Influenza Virus A ◽  
B Virus

Due to the common symptoms of COVID-19, patients are similar to influenza-like illness. Therefore, the detection method would be crucial to discriminate between SARS-CoV-2 and influenza virus-infected patients. In this study, CRISPR-Cas12a-based detection was applied for detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza A virus, and influenza B virus which would be a practical and attractive application for screening of patients with COVID-19 and influenza in areas with limited resources. The limit of detection for SARS-CoV-2, influenza A, and influenza B detection was 10, 103, and 103 copies/reaction, respectively. Moreover, the assays yielded no cross-reactivity against other respiratory viruses. The results revealed that the detection of influenza virus and SARS-CoV-2 by using RT-RPA and CRISPR-Cas12a technology reaches 96.23% sensitivity and 100% specificity for SARS-CoV-2 detection. The sensitivity for influenza virus A and B detections was 85.07% and 94.87%, respectively. In addition, the specificity for influenza virus A and B detections was approximately 96%. In conclusion, the RT-RPA with CRISPR-Cas12a assay was an effective method for the screening of influenza viruses and SARS-CoV-2 which could be applied to detect other infectious diseases in the future.

scholarly journals Bioluminescence-Based Neuraminidase Inhibition Assay for Monitoring Influenza Virus Drug Susceptibility in Clinical Specimens

2013 ◽  
Vol 57 (11) ◽  
pp. 5209-5215 ◽  
Author(s):  
Henju Marjuki ◽  
Vasiliy P. Mishin ◽  
Katrina Sleeman ◽  
Margaret Okomo-Adhiambo ◽  
Tiffany G. Sheu ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Point Of Care ◽  
Drug Susceptibility ◽  
Influenza Viruses ◽  
Clinical Specimens ◽  
Influenza Virus A ◽  
Oseltamivir Resistance ◽  
Resistance Markers ◽  
The Difference ◽  
Inhibitor Resistance

ABSTRACTThe QFlu prototype bioluminescence-based neuraminidase (NA) inhibition (NI) assay kit was designed to detect NA inhibitor (NAI)-resistant influenza viruses at point of care. Here, we evaluated its suitability for drug susceptibility assessment at a surveillance laboratory. A comprehensive panel of reference viruses (n= 14) and a set of 90 seasonal influenza virus A and B isolates were included for testing with oseltamivir and/or zanamivir in the QFlu assay using the manufacturer-recommended protocol and a modified version attuned to surveillance requirements. The 50% inhibitory concentrations (IC50s) generated were compared with those of NI assays currently used for monitoring influenza drug susceptibility, the fluorescent (FL) and chemiluminescent (CL) assays. To provide proof of principle, clinical specimens (n= 235) confirmed by real-time reverse transcription (RT)-PCR to contain influenza virus A(H1N1)pdm09 and prescreened for the oseltamivir resistance marker H275Y using pyrosequencing were subsequently tested in the QFlu assay. All three NI assays were able to discriminate the reference NA variants and their matching wild-type viruses based on the difference in their IC50s. Unless the antigenic types were first identified, certain NA variants (e.g., H3N2 with E119V) could be detected among seasonal viruses using the FL assays only. Notably, the QFlu assay identified oseltamivir-resistant A(H1N1)pdm09 viruses carrying the H275Y marker directly in clinical specimens, which is not feasible with the other two phenotypic assays, which required prior virus culturing in cells. Furthermore, The QFlu assay allows detection of the influenza virus A and B isolates carrying established and potential NA inhibitor resistance markers and may become a useful tool for monitoring drug resistance in clinical specimens.

scholarly journals The Multivalent PDZ Domain-containing Protein PDZK1 Regulates Transport Activity of Renal Urate-Anion Exchanger URAT1 via Its C Terminus

2004 ◽  
Vol 279 (44) ◽  
pp. 45942-45950 ◽  
Author(s):  
Naohiko Anzai ◽  
Hiroki Miyazaki ◽  
Rie Noshiro ◽  
Suparat Khamdang ◽  
Arthit Chairoungdua ◽  
...  
Keyword(s):  
Anion Exchanger ◽  
Apical Membrane ◽  
Organic Anion ◽  
Pdz Domain ◽  
Hek293 Cells ◽  
Transport Activity ◽  
Yeast Two Hybrid ◽  
Urate Transport ◽  
Anion Transporter ◽  
Two Hybrid

The urate-anion exchanger URAT1 is a member of the organic anion transporter (OAT) family that regulates blood urate level in humans and is targeted by uricosuric and antiuricosuric agents (Enomoto, A., Kimura, H., Chairoungdua, A., Shigeta, Y., Jutabha, P., Cha, S. H., Hosoyamada, M., Takeda, M., Sekine, T., Igarashi, T., Matsuo, H., Kikuchi, Y., Oda, T., Ichida, K., Hosoya, T., Shimotaka, K., Niwa, T., Kanai, Y., and Endou, H. (2002)Nature417, 447–452). URAT1 is expressed only in the kidney, where it is thought to participate in tubular urate reabsorption. We found that the multivalent PDZ (PSD-95,Drosophiladiscs-large protein,Zonula occludensprotein 1) domain-containing protein, PDZK1 interacts with URAT1 in a yeast two-hybrid screen. Such an interaction requires the PDZ motif of URAT1 in its extreme intracellular C-terminal region and the first, second, and fourth PDZ domains of PDZK1 as identified by yeast two-hybrid assay,in vitrobinding assay and surface plasmon resonance analysis (KD= 1.97–514 nm). Coimmunoprecipitation studies revealed that the wild-type URAT1, but not its mutant lacking the PDZ-motif, directly interacts with PDZK1. Colocalization of URAT1 and PDZK1 was observed at the apical membrane of renal proximal tubular cells. The association of URAT1 with PDZK1 enhanced urate transport activities in HEK293 cells (1.4-fold), and the deletion of the URAT1 C-terminal PDZ motif abolished this effect. The augmentation of the transport activity was accompanied by a significant increase in theVmaxof urate transport via URAT1 and was associated with the increased surface expression level of URAT1 protein from HEK293 cells stably expressing URAT1 transfected with PDZK1. Taken together, the present study indicates the novel role of PDZK1 in regulating the functional activity of URAT1-mediated urate transport in the apical membrane of renal proximal tubules.

scholarly journals Different subtypes of influenza A viruses target different human proteins and pathways leading to different pathogenic phenotypes

2019 ◽  
Author(s):  
Yujie Wang ◽  
Ting Song ◽  
Kaiwu Li ◽  
Yuan Jin ◽  
Junjie Yue ◽  
...  
Keyword(s):  
Influenza A ◽  
Influenza Viruses ◽  
Host Protein ◽  
Yeast Two Hybrid ◽  
Influenza A Viruses ◽  
Protein Protein Interaction ◽  
Ppi Networks ◽  
Tnf Α ◽  
Human Proteins ◽  
Two Hybrid

AbstractDifferent subtypes of Influenza A viruses cause different pathogenic phenotypes after infecting human bodies. Direct binary interactions between viral proteins and human proteins provide an important background for influenza viruses to cause complex pathologies of hosts. Here, we demonstrated the different impacts on the TNF-α-induced NF-κB activation of H1N1 and H5N1 virus proteins. By further examining the virus-host protein-protein interactions (PPI), we found that the same segment protein of the H1N1 and H5N1 viruses target on different host proteins. We then performed a yeast two-hybrid analysis of a highly pathogenic avian H5N1 influenza virus and human proteins. Influenza-host protein-protein interaction networks of three strains of influenza A viruses (including two other reported influenza-host PPI networks) were systematically compared and mapped on the network level and the pathway level. The results show subtype-specific characters of the influenza-host protein interactome, which may response for the specific pathogenic mechanisms of different subtypes of influenza viruses.ImportanceInfluenza A virus (IAV) can cause contagious respiratory illness, namely influenza (flu). The symptoms of infections from different subtypes of IAVs vary from mild to severe illness. The mechanism of these different pathogenic phenotypes remains poorly understood. Our results show that the same NA and NP segments from H1N1 and H5N1 virus cause different impacts on the TNF-α-induced NF-κB pathway. Furthermore, we generated a yeast two-hybrid protein-protein interaction (PPI) network between H5N1 and human proteins. By systematically comparing the influenza-host PPI networks of three strains of IAVs, we show that different subtypes of IAVs target different human proteins and pathways, which may have led to different pathogenic phenotypes.

Use of GFP-expressing influenza viruses for the detection of influenza virus A/H5N1 neutralizing antibodies

Vaccine ◽  
2011 ◽  
Vol 29 (18) ◽  
pp. 3424-3430 ◽  
Author(s):  
Guus F. Rimmelzwaan ◽  
R. Joyce Verburgh ◽  
Nella J. Nieuwkoop ◽  
Theo M. Bestebroer ◽  
Ron A.M. Fouchier ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Neutralizing Antibodies ◽  
Influenza Viruses ◽  
Influenza Virus A

scholarly journals Interferon Lambda Delays the Emergence of Influenza Virus Resistance to Oseltamivir

2021 ◽  
Vol 9 (6) ◽  
pp. 1196
Author(s):  
Chiara Medaglia ◽  
Arnaud Charles-Antoine Zwygart ◽  
Paulo Jacob Silva ◽  
Samuel Constant ◽  
Song Huang ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Neuraminidase Inhibitor ◽  
Influenza Viruses ◽  
Human Interferon ◽  
Infection Model ◽  
Single Mutation ◽  
Resistance Mutations ◽  
Species Barrier ◽  
Interferon Lambda

Influenza viruses are a leading cause of morbidity and mortality worldwide. These air-borne pathogens are able to cross the species barrier, leading to regular seasonal epidemics and sporadic pandemics. Influenza viruses also possess a high genetic variability, which allows for the acquisition of resistance mutations to antivirals. Combination therapies with two or more drugs targeting different mechanisms of viral replication have been considered an advantageous option to not only enhance the effectiveness of the individual treatments, but also reduce the likelihood of resistance emergence. Using an in vitro infection model, we assessed the barrier to viral resistance of a combination therapy with the neuraminidase inhibitor oseltamivir and human interferon lambda against the pandemic H1N1 A/Netherlands/602/2009 (H1N1pdm09) virus. We serially passaged the virus in a cell line derived from human bronchial epithelial cells in the presence or absence of increasing concentrations of oseltamivir alone or oseltamivir plus interferon lambda. While the treatment with oseltamivir alone quickly induced the emergence of antiviral resistance through a single mutation in the neuraminidase gene, the co-administration of interferon lambda delayed the emergence of drug-resistant influenza virus variants. Our results suggest a possible clinical application of interferon lambda in combination with oseltamivir to treat influenza.

scholarly journals Development of DNA-Biochip for Identification of Influenza A Virus Subtypes

2012 ◽  
pp. 89-93
Author(s):  
A. N. Shikov ◽  
E. I. Sergeeva ◽  
O. K. Demina ◽  
V. A. Ternovoy ◽  
V. V. Ryabinin ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Influenza A Virus ◽  
Influenza A ◽  
Seasonal Influenza ◽  
Highly Pathogenic Avian Influenza ◽  
Influenza Viruses ◽  
Highly Pathogenic ◽  
Influenza Virus A ◽  
Pathogenic Avian Influenza ◽  
Pathogenic Avian Influenza Virus

Developed was the DNA-biochip to identify subtypes of influenza A virus, pathogenic for humans. Microchip was capable of detecting H1, H3, H5-subtypes of hemagglutinin (including H1-subtype of pandemic A/H1N1(2009) influenza virus ) and neuraminidase subtypes N1,N2 of influenza virus. This microchip was successfully tested on the strains of A/H5N1 highly pathogenic avian influenza virus, A/H1N1(2009) pandemic influenza virus, A/H1N1 and A/H3N2 seasonal influenza viruses.

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