Clonogenic assay of type a influenza viruses reveals noninfectious cell-killing (apoptosis-inducing) particles

Journal of Virology
John M NgunjiriPhilip I Marcus

Abstract

Clonogenic (single-cell plating) assays were used to define and quantify subpopulations of two genetically closely related variants of influenza virus A/TK/OR/71 that differed primarily in the size of the NS1 gene product; they expressed a full-size (amino acids [aa] 1 to 230) or truncated (aa 1 to 124) NS1 protein. Monolayers of Vero cells were infected with different amounts of virus, monodispersed, and plated. Cell survival curves were generated from the fraction of cells that produced visible colonies as a function of virus multiplicity. The exponential loss of colony-forming capacity at low multiplicities demonstrated that a single virus particle sufficed to kill a cell. The ratios of cell-killing particles (CKP) to plaque-forming particles (PFP) were 1:1 and 7:1 in populations of variants NS1(1-124) and NS1(1-230), respectively. This study revealed a new class of particles in influenza virus populations-noninfectious CKP. Both infectious and noninfectious CKP were 6.3 times more resistant to UV radiation than PFP activity. Based on UV target theory, a functional polymerase subunit was implicated in a rate-limiting step in cell killing. Since influenza viruses kill cells by apoptosis (programmed cell death), CKP are functi...Continue Reading

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Citations

Mar 6, 2012·Journal of Interferon & Cytokine Research : the Official Journal of the International Society for Interferon and Cytokine Research·John M NgunjiriPhilip I Marcus
Dec 12, 2012·Journal of Interferon & Cytokine Research : the Official Journal of the International Society for Interferon and Cytokine Research·John M NgunjiriPhilip I Marcus
Nov 8, 2012·Journal of Interferon & Cytokine Research : the Official Journal of the International Society for Interferon and Cytokine Research·Christopher P Malinoski, Philip I Marcus
Jan 4, 2013·Journal of Virology·Christopher B BrookeJonathan W Yewdell
Sep 29, 2011·PloS One·Heather MacInnesFrederick Koster
Jul 30, 2014·Future Virology·Christopher B Brooke
May 7, 2014·Trends in Microbiology·Marie GerberRoland Marquet
May 21, 2013·Journal of Virological Methods·Danielle H W VleckenLeo A van der Pol
Dec 6, 2011·Journal of Interferon & Cytokine Research : the Official Journal of the International Society for Interferon and Cytokine Research·Christopher P Malinoski, Philip I Marcus
Oct 8, 2013·PloS One·Yo Han JangBaik Lin Seong
Sep 1, 2017·Journal of Virology·Christopher B Brooke
Sep 4, 2019·Annual Review of Animal Biosciences·Amir GhorbaniChang-Won Lee
Feb 2, 2019·Scientific Reports·Kazuhito V TabataHiroyuki Noji

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Apoptosis

Apoptosis is a specific process that leads to programmed cell death through the activation of an evolutionary conserved intracellular pathway leading to pathognomic cellular changes distinct from cellular necrosis