We used embryo fibroblasts from Mx2-Luc transgenic mice that express Firefly luciferase under control of the interferon (IFN)-regulated mouse Mx2 promoter to develop simple nonviral bioassays for type I and type III IFN. Since type III IFN is acid-labile, Mx2-Luc fibroblasts detected the presence of type I IFN in acid-treated biological samples with high sensitivity and selectivity. For selective detection of type III IFN, we employed embryo fibroblasts from Mx2-Luc mutant mice that lack functional receptors for type I IFN. The sensitivity of this latter assay remained comparatively low, presumably because type III IFN receptors are not abundantly present on fibroblasts. The main advantages of our novel IFN assays are that they are easy to perform, yield fast results, and can be used in laboratories that are not licensed for work with infectious agents. Further, the type I IFN assay has superior sensitivity than commercially available enzyme-linked immunosorbent assay systems.
Get full access to this article
View all access options for this article.
References
1.
AnkN, IversenMB, BartholdyC, StaeheliP, HartmannR, JensenUB, Dagnaes-HansenF, ThomsenAR, ChenZ, HaugenH, KlucherK, PaludanSR. 2008. An important role for type III interferon (IFN-lambda/IL-28) in TLR-induced antiviral activity. J Immunol, 180,4:2474–2485.
2.
AnkN, PaludanSR. 2009. Type III IFNs: new layers of complexity in innate antiviral immunity. Biofactors, 35,1:82–87.
3.
BalkwillFR, GriffinDB, BandHA, BeverleyPC. 1983. Immune human lymphocytes produce an acid-labile alpha-interferon. J Exp Med, 157,3:1059–1063.
4.
BouloyM, JanzenC, VialatP, KhunH, PavlovicJ, HuerreM, HallerO. 2001. Genetic evidence for an interferon-antagonistic function of rift valley fever virus nonstructural protein NSs. J Virol, 75,3:1371–1377.
5.
CanosiU, MasciaM, GazzaL, Serlupi-CrescenziO, DoniniS, AntonettiF, GalliG. 1996. A highly precise reporter gene bioassay for type I interferon. J Immunol Methods, 199,1:69–76.
6.
ChiB, DickensheetsHL, SpannKM, AlstonMA, LuongoC, DumoutierL, HuangJ, RenauldJC, KotenkoSV, RoedererM, BeelerJA, DonnellyRP, CollinsPL, RabinRL. 2006. Alpha and lambda interferon together mediate suppression of CD4 T cells induced by respiratory syncytial virus. J Virol, 80,10:5032–5040.
7.
DellgrenC, GadHH, HammingOJ, MelchjorsenJ, HartmannR. 2009. Human interferon-lambda3 is a potent member of the type III interferon family. Genes Immun, 10,2:125–131.
8.
DumoutierL, TounsiA, MichielsT, SommereynsC, KotenkoSV, RenauldJC. 2004. Role of the interleukin (IL)-28 receptor tyrosine residues for antiviral and antiproliferative activity of IL-29/interferon-lambda 1: similarities with type I interferon signaling. J Biol Chem, 279,31:32269–32274.
9.
FrancoisC, BernardI, CastelainS, CharlestonB, FrayMD, CapiodJC, DuverlieG. 2005. Quantification of different human alpha interferon subtypes and pegylated interferon activities by measuring MxA promoter activation. Antimicrob Agents Chemother, 49,9:3770–3775.
10.
GadHH, DellgrenC, HammingOJ, VendsS, PaludanSR, HartmannR. 2009. Interferon-lambda is functionally an interferon but structurally related to the interleukin-10 family. J Biol Chem, 284,31:20869–20875.
11.
HallerO, KochsG, WeberF. 2006. The interferon response circuit: induction and suppression by pathogenic viruses. Virology, 344,1:119–130.
HallerO, StaeheliP, KochsG. 2009. Protective role of interferon-induced Mx GTPases against influenza viruses. Rev Sci Tech, 28,1:219–231.
14.
KochsG, KoernerI, ThielL, KothlowS, KaspersB, RuggliN, SummerfieldA, PavlovicJ, StechJ, StaeheliP. 2007. Properties of H7N7 influenza A virus strain SC35M lacking interferon antagonist NS1 in mice and chickens. J Gen Virol, 88,Pt 5:1403–1409.
15.
KoernerI, KochsG, KalinkeU, WeissS, StaeheliP. 2007. Protective role of beta interferon in host defense against influenza A virus. J Virol, 81,4:2025–2030.
16.
KotenkoSV, GallagherG, BaurinVV, Lewis-AntesA, ShenM, ShahNK, LangerJA, SheikhF, DickensheetsH, DonnellyRP. 2003. IFN-lambdas mediate antiviral protection through a distinct class II cytokine receptor complex. Nat Immunol, 4,1:69–77.
17.
Levy-KoenigRE, GolgherRR, PauckerK. 1970. Immunology of interferons. II. Heterospecific activities of human interferons and their neutralization by antibody. J Immunol, 104,4:791–797.
18.
LienenklausS, CornitescuM, ZietaraN, LyszkiewiczM, GekaraN, JablonskaJ, EdenhoferF, RajewskyK, BruderD, HafnerM, StaeheliP, WeissS. 2009. Novel reporter mouse reveals constitutive and inflammatory expression of IFN-beta in vivo. J Immunol, 183,5:3229–3236.
19.
MeagerA. 2002. Biological assays for interferons. J Immunol Methods, 261,1–2:21–36.
20.
MordsteinM, KochsG, DumoutierL, RenauldJC, PaludanSR, KlucherK, StaeheliP. 2008. Interferon-lambda contributes to innate immunity of mice against influenza A virus but not against hepatotropic viruses. PLoS Pathog, 4,9:e1000151.
21.
PestkaS. 1986. Interferon standards and general abbreviations. Methods Enzymol, 119:14–23.
22.
PulvererJE, RandU, LienenklausS, KugelD, ZietaraN, KochsG, NaumannR, WeissS, StaeheliP, HauserH, KosterM. 2010. Temporal and spatial resolution of type I and III IFN responses in vivo. J Virol, 84,17:8626–8638.
23.
SchwarzH, HarlinO, OhnemusA, KaspersB, StaeheliP. 2004. Synthesis of IFN-beta by virus-infected chicken embryo cells demonstrated with specific antisera and a new bioassay. J Interferon Cytokine Res, 24,3:179–184.
24.
SommereynsC, PaulS, StaeheliP, MichielsT. 2008. IFN-lambda (IFN-lambda) is expressed in a tissue-dependent fashion and primarily acts on epithelial cells in vivo. PLoS Pathog, 4,3:e1000017.
25.
ZhouZ, HammingOJ, AnkN, PaludanSR, NielsenAL, HartmannR. 2007. Type III interferon (IFN) induces a type I IFN-like response in a restricted subset of cells through signaling pathways involving both the Jak-STAT pathway and the mitogen-activated protein kinases. J Virol, 81,14:7749–7758.
