In this report we describe the generation of a mouse monoclonal antibody (MAb) against the influenza A virus PB1-F2 protein that is derived from a +1 reading frame of the polymerase basic protein (PB1) gene segment. We further present data that the hybridoma subclone F2-6G10 produces antibodies that specifically recognize the PB1-F2 protein of H1N1 influenza virus types only. The antibody can be used for immunodetection of the PB1-F2 protein in ELISA, Western blot, immunoprecipitation, and immunofluorescence assays.
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References
1.
NeumannG, BrownleeGG, FodorG, KawaokaY. Orthomyxovirus replication, transcription, and polyadenylation. Curr Top Microbiol Immunol, 2004; 283:121–143.
2.
GabrielG, DauberB, WolffT, PlanzO, KlenkHD, StechJ. The viral polymerase mediates adaptation of an avian influenza virus to a mammalian host. Proc Natl Acad Sci USA, 2005; 102:18590–18595.
3.
DigardP, BlokVC, InglisSC. Complex formation between influenza virus polymerase proteins expressed in Xenopus oocytes. Virology, 1989; 171:162–169.
4.
ChenW, CalvoPA, MalideD, GibbsJ, SchubertU, BacikI, BastaS, O'NeillR, SchickliJ, PaleseP, HenkleinP, BenninkJR, YewdellJW. A novel influenza A virus mitochondrial protein that induces cell death. Nat Med, 2001; 7:1306–1312.
5.
MazurI, AnhlanD, MitznerD, WixlerL, SchubertU, LudwigS. The proapoptotic influenza A virus protein PB1-F2 regulates viral polymerase activity by interaction with the PB1 protein. Cell Microbiol, 2008; 10:1140–1152.
6.
ZamarinD, Garcia-SastreA, XiaoX, WangR, PaleseP. Influenza virus PB1-F2 protein induces cell death through mitochondrial ANT3 and VDAC1. PLoS Pathog, 2005; 1:e4.
7.
McAuleyJL, HornungF, BoydKL, SmithAM, McKeonR, BenninkJ, YewdellJW, McCullersJA. Expression of the 1918 influenza A virus PB1-F2 enhances the pathogenesis of viral and secondary bacterial pneumonia. Cell Host Microbe, 2007; 2:240–249.
8.
MitznerD, DudekSE, StudtruckerN, AnhlanD, MazurI, WissingJ, JanschL, WixlerL, BrunsK, SharmaA, WrayA, HenkleinP, LudwigS, SchubertU. Phosphorylation of the influenza A virus protein PB1-F2 by PKC is crucial for apoptosis promoting functions in monocytes. Cell Microbiol, 2009; 11:1502–1516.
9.
ConenelloGM, ZamarinD, PerroneLA, TumpeyT, PaleseP. A single mutation in the PB1-F2 of H5N1 (HK/97) and 1918 influenza A viruses contributes to increased virulence. PLoS Pathog, 2007; 3:1414–1421.
10.
BaslerCF, WangX, MuhlbergerE, VolchkovV, ParagasJ, KlenkHD, Garcia-SastreA, PaleseP. The Ebola virus VP35 protein functions as a type I IFN antagonist. Proc Natl Acad Sci USA, 2000; 97:12289–12294.
