scholarly journals A High Throughput Assay for Screening Host Restriction Factors and Antivirals Targeting Influenza A Virus

2016 ◽  
Vol 7 ◽  
Author(s):  
Lingyan Wang ◽  
Wenjun Li ◽  
Shitao Li
Keyword(s):  
Influenza A Virus ◽  
High Throughput ◽  
Influenza A ◽  
Restriction Factors ◽  
Host Restriction ◽  
Host Restriction Factors ◽  
High Throughput Assay

Amphotericin b effect on the sensitivity to influenza infection of WI-38 VA-13 cells with IFITM3 gene knockout

2021 ◽  
Vol 21 (3) ◽  
pp. 109-112
Author(s):  
Kira S. Koryabina ◽  
Mariya V. Sergeeva ◽  
Andrey B. Komissarov ◽  
Nataliya V. Eshchenko ◽  
Grigoriy A. Stepanov
Keyword(s):  
Amphotericin B ◽  
Influenza A Virus ◽  
Influenza A ◽  
Gene Knockout ◽  
Influenza Infection ◽  
Restriction Factors ◽  
Host Restriction ◽  
Host Restriction Factors ◽  
Infected Cells ◽  
The Difference

BACKGROUND: The application of CRISPR/Cas9 is one of the most rapidly developing areas in biotechnology. This method was used to obtain clones of а human origin cell line with knockout of one or more genes of the IFITM family, representing host restriction factors for influenza infection. Amphotericin B has previously been shown to promote influenza infection by blocking IFITM3 function. AIM: The aim of this study was to evaluate the effect of amphotericin B on the sensitivity of IFITM knockout cells to influenza A virus infection. MATERIALS AND METHODS: WI-38 VA-13 cells and mutant clones with IFITM3 knockout (F3 clone) or IFITM1, IFITM3 knockout (clone E12) were infected with influenza virus A/PR/8/34 (H1N1) in the presence or absence of amphotericin B. Forty-four hours after infection, the culture medium was taken to determine the infectious activity of the virus by titration in the MDCK cell culture, as well as the hemagglutinating activity of the virus. The infected cells were stained with fluorescently labeled antibodies against the viral NP protein, and the number of NP-positive cells was determined by flow cytometry. RESULTS: The addition of amphotericin B increased the hemagglutinating and infectious activity of the virus in WI-38 VA-13cells, while the difference was insignificant for clones with IFITM gene knockout. A similar dependency was obtained for the percent of infected cells. CONCLUSIONS: Mutant cells with a knockout of one or several genes of the IFITM family were equally susceptible to influenza infection regardless of the addition of amphotericin B, which confirms the crucial importance of a defect in the IFITM3 protein in increasing the permissiveness of cells to influenza A virus.

scholarly journals Establishment of a High-Throughput Assay to Monitor Influenza A Virus RNA Transcription and Replication

PLoS ONE ◽  
2015 ◽  
Vol 10 (7) ◽  
pp. e0133558 ◽  
Author(s):  
Zhen Wang ◽  
Fei Zhao ◽  
Qian Gao ◽  
Zhenlong Liu ◽  
Yongxin Zhang ◽  
...  
Keyword(s):  
Influenza A Virus ◽  
High Throughput ◽  
Influenza A ◽  
Rna Transcription ◽  
Virus Rna ◽  
High Throughput Assay ◽  
Transcription And Replication

Sequence analysis identified an HTLV-1 subtype and mutations of host restriction factors susceptible to HAM/TSP

2017 ◽  
Vol 381 ◽  
pp. 1006
Author(s):  
S. Nozuma ◽  
E. Matsuura ◽  
T. Matsuzaki ◽  
D. Kodama ◽  
R. Kubota ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Restriction Factors ◽  
Host Restriction ◽  
Host Restriction Factors

Host Restriction Factors and Human Immunodeficiency Virus (HIV-1): A Dynamic Interplay Involving All Phases of the Viral Life Cycle

2018 ◽  
Vol 16 (3) ◽  
pp. 184-207 ◽  
Author(s):  
Vanessa D`Urbano ◽  
Elisa De Crignis ◽  
Maria Carla Re
Keyword(s):  
Mammalian Cells ◽  
Viral Evolution ◽  
Type I ◽  
Restriction Factors ◽  
Host Restriction ◽  
Host Restriction Factors ◽  
Immunodeficiency Virus ◽  
Lentiviral Infection ◽  
Specific Proteins ◽  
Hiv 1

Mammalian cells have evolved several mechanisms to prevent or block lentiviral infection and spread. Among the innate immune mechanisms, the signaling cascade triggered by type I interferon (IFN) plays a pivotal role in limiting the burden of HIV-1. In the presence of IFN, human cells upregulate the expression of a number of genes, referred to as IFN-stimulated genes (ISGs), many of them acting as antiviral restriction factors (RFs). RFs are dominant proteins that target different essential steps of the viral cycle, thereby providing an early line of defense against the virus. The identification and characterization of RFs have provided unique insights into the molecular biology of HIV-1, further revealing the complex host-pathogen interplay that characterizes the infection. The presence of RFs drove viral evolution, forcing the virus to develop specific proteins to counteract their activity. The knowledge of the mechanisms that prevent viral infection and their viral counterparts may offer new insights to improve current antiviral strategies. This review provides an overview of the RFs targeting HIV-1 replication and the mechanisms that regulate their expression as well as their impact on viral replication and the clinical course of the disease.

scholarly journals Dysregulation of M segment gene expression contributes to influenza A virus host restriction

2019 ◽  
Vol 15 (8) ◽  
pp. e1007892 ◽  
Author(s):  
Brenda M. Calderon ◽  
Shamika Danzy ◽  
Gabrielle K. Delima ◽  
Nathan T. Jacobs ◽  
Ketaki Ganti ◽  
...  
Keyword(s):  
Gene Expression ◽  
Influenza A Virus ◽  
Influenza A ◽  
Host Restriction

scholarly journals Epigenetic factor siRNA screen during primary KSHV infection identifies novel host restriction factors for the lytic cycle of KSHV

2020 ◽  
Vol 16 (1) ◽  
pp. e1008268 ◽  
Author(s):  
Nenavath Gopal Naik ◽  
Thomas Hong Nguyen ◽  
Lauren Roberts ◽  
Luke Todd Fischer ◽  
Katherine Glickman ◽  
...  
Keyword(s):  
Lytic Cycle ◽  
Restriction Factors ◽  
Sirna Screen ◽  
Kshv Infection ◽  
Host Restriction ◽  
Host Restriction Factors ◽  
Epigenetic Factor ◽  
Novel Host

scholarly journals Objective pathogen monitoring in nursery and finisher pigs by monthly laboratory diagnostic testing

2020 ◽  
Vol 6 (1) ◽  
Author(s):  
Nicole B. Goecke ◽  
Maja Kobberø ◽  
Thomas K. Kusk ◽  
Charlotte K. Hjulsager ◽  
Ken Steen Pedersen ◽  
...  
Keyword(s):  
Real Time ◽  
San Francisco ◽  
Influenza A Virus ◽  
High Throughput ◽  
Real Time Pcr ◽  
Influenza A ◽  
Clinical Signs ◽  
Swine Influenza ◽  
Porcine Cytomegalovirus ◽  
Swine Influenza A Virus

Abstract Background Infectious diseases are of great economic importance in commercial pig production, causing both clinical and subclinical disease, with influence on welfare, productivity, and antibiotic use. The causes of these diseases are often multifactorial and laboratory diagnostics are seldom routinely performed. The aim of the present study was to explore the benefits of monthly pathogen monitoring in nursery and finisher herds and to examine association between laboratory results and observed clinical signs, including coughing and diarrhoea. Three monthly samplings were conducted in three different age groups in six nursery and four finisher production units. For each unit, two pens were randomly selected in each age group and evaluated for coughing and diarrhoea events. Furthermore, faecal sock and oral fluid samples were collected in the selected pens and analysed for 18 respiratory and enteric viral and bacterial pathogens using the high-throughput real-time PCR BioMark HD platform (Fluidigm, South San Francisco, USA). Results In total, 174 pens were sampled in which eight coughing events and 77 diarrhoeic events were observed. The overall findings showed that swine influenza A virus, porcine circovirus 2, porcine cytomegalovirus, Brachyspira pilosicoli, Lawsonia intracellularis, Escherichia coli fimbria types F4 and F18 were found to be prevalent in several of the herds. Association between coughing events and the presence of swine influenza A virus, porcine cytomegalovirus (Cq ≤ 20) or a combination of these were found. Furthermore, an association between diarrhoeic events and the presence of L. intracellularis (Cq ≤ 24) or B. pilosicoli (Cq ≤ 26) was found. Conclusions The use of high-throughput real-time PCR analysis for continuous monitoring of pathogens and thereby dynamics of infections in a pig herd, provided the veterinarian and farmer with an objective knowledge on the distribution of pathogens in the herd. In addition, the use of a high-throughput method in combination with information about clinical signs, productivity, health status and antibiotic consumption, presents a new and innovative way of diagnosing and monitoring pig herds and even to a lower cost than the traditional method.

scholarly journals Identification of novel influenza A virus exposures by an improved high‐throughput multiplex MAGPIX platform and serum adsorption

2019 ◽  
Vol 14 (2) ◽  
pp. 129-141
Author(s):  
Zhu‐Nan Li ◽  
Emily Cheng ◽  
Eugenie Poirot ◽  
Kimberly M. Weber ◽  
Paul Carney ◽  
...  
Keyword(s):  
Influenza A Virus ◽  
High Throughput ◽  
Influenza A ◽  
Novel Influenza A

scholarly journals A pooled genome-wide screening strategy to identify and rank influenza host restriction factors in cell-based vaccine production platforms

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
David M. Sharon ◽  
Sean Nesdoly ◽  
Hsin J. Yang ◽  
Jean-François Gélinas ◽  
Yu Xia ◽  
...  
Keyword(s):  
Screening Strategy ◽  
Vaccine Production ◽  
Restriction Factors ◽  
Host Restriction ◽  
Host Restriction Factors ◽  
Genome Wide
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