We have recently described the production of antisera which react with the human herpesvirus 6 (HHV-6) major capsid protein (MCP). We describe here the use of these specific antisera to characterize the kinetics of expression of the HHV-6 MCP and demonstrate that it is a late virus protein, expressed after DNA replication. We have used the expression of this protein as a measure of the efficacy of the antiviral nucleoside analogues 9-(2-hydroxyethoxymethyl)guanine (ACV) and 9-(1,3-dihydroxy-2-propoxymethyl)guanine (DHPG) upon HHV-6 replication and have determined 50% inhibitory concentrations to be 150 μm and 2 μm, respectively.
References
1.
AblashiD.V.JosephsD.F.BuchbinderA.HellmanK.NakamuraS.LianaT.LussoP.KaplanM.DahlbergJ.MemonS.ImamF.AblashiK.L.MarkhamP.D.KramarskyB.KruegerG.R.F.BiberfeldP.Wong-StaalF.SalahuddinS.Z., and GalloR.C. (1988) Human B-lymphotropic virus (human herpesvirus-6). J Virol Meth21: 29–48.
2.
AgutH.HurauxJ.M.CollandreH., and MontagnierL. (1989) Susceptibility of human herpesvirus 6 to acyclovir and ganciclovir. Lancetii: 626.
3.
BironK.K.StanatS.C.SorrellJ.B.FyfeJ.A.KellerP.M.LambeC.U., and NelsonD.J. (1985) Metabolic activation of the nucleoside analog 9-{[2-hydroxy-1-(hydroxymethyl) ethoxyl]methyl}guanine in human diploid fibroblasts infected with human cytomegalovirus. Proc Natl Acad Sci USA82: 2473–2477.
4.
ChengY.-C.HuangE.-S.LinJ.-C.MarE.-C.PaganoJ.S.DutschmanG.E., and GrillS.P. (1983) Unique spectrum of activity of 9-[(1,3-dihydroxy-2-propoxy)methyl]-guanine against herpesviruses in vitro and its mode of action against herpes simplex virus type 1. Proc Natl Acad Sci USA80: 2767–2770.
5.
ColbyB.M.ShawJ.E.ElionG.B., and PaganoJ.S. (1980) Effect of acyclovir [9-(2-hydroxyethoxymethyl)guanine] on Epstein–Barr virus DNA replication. J Virol34: 560–568.
6.
ElionG.B. (1982) Mechanism of action and selectivity of acyclovir. Am J Med73: 7–13.
7.
FurmanP.A., and McGuirtP.V. (1983) Effect of acyclovir on viral protein synthesis in cells infected with herpes simplex virus type 1. Antimicrob Agents Chemother23: 332–334.
8.
GeballeA.P.LeachF.S., and MocarskiE.S. (1986) Regulation of cytomegalovirus late gene expression: genes are controlled by post-transcriptional events. J Virol57: 864–874.
9.
GranlundD.J., and PearsonG.R. (1977) Membrane antigen expression in Epstein–Barr virus-infected Raji cells in the presence of phosphoacetic acid. Virology83: 217–220.
10.
HorvatR.T.WoodC., and BalachandranN. (1989) Transactivation of human immunodeficiency virus promoter by human herpesvirus 6. J Virol63: 970–973.
11.
JarrettR.F.GledhillS.QureshiF.CraeS.H.MadhokR.BrownI.EvansI.KrajewskiA.O'BrienC.J.CartwrightR.A.VenablesP., and OnionsD.E. (1988) Identification of human herpesvirus 6-specific DNA sequences in two patients with non-Hodgkin's lymphoma. Leukemia2: 496–502.
12.
JohnsonD.A.GautschJ.W.SportsmanJ.R., and ElderJ.H. (1984) Improved technique utilizing non-fat dry milk for analysis of proteins and nucleic acids transferred to nitrocellulose. Gene Anal Tech1: 3–8.
13.
JosephsS.F.BuchbinderA.StreicherH.Z.AblashiD.V.SalahuddinS.Z.GuoH.-G.Wong-StaalF.CossmanJ.RaffeldM.SundeenJ.LevineP.BiggarR.KruegerG.R.F.FoxR.I., and GalloR.C. (1988) Detection of human B-lymphotropic virus (human herpesvirus 6) sequences in B cell lymphoma tissues of three patients. Leukemia2: 132–135.
14.
KishiM.HaradaH.TakahashiM.TanakaA.HayashiM.NonayamaM.JosephsS.F.BuchbinderA.SchachterF.AblashiD.V.Wong-StaalF.SalahuddinS.Z., and GalloR.C. (1988) A repeat sequence, GGGTTA, is shared by DNA of human herpesvirus 6 and Marke's disease virus. J Virol62: 4824–4827.
15.
KruegerG.R.F.KochB., and AblashiD.V. (1988) Persistent fatigue and depression in patient with antibody to human B-lymphotropic virus. Lancetii: 36.
16.
LittlerE.LawrenceG.LiuM.-Y.BarrellB.G., and ArrandJ.R. (1990) Identification, cloning and expression of the major capsid protein gene of human herpesvirus type 6. J Virol64: 714–722.
17.
LussoP.EnsoliB.MarkhamP.D.AblashiD.V.SalahuddinS.Z.TschachlerE.Wong-StaalF., and GalloR.C. (1989) Productive dual infection of human CD4+ T lymphocytes by HIV-1 and HHV-6. Nature337: 370–373.
18.
ManiatisT.FritschE.F., and SambrookJ. (1982) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
19.
RusslerS.K.TapperM.A., and CarriganD.R. (1989) Susceptibility of human herpesvirus 6 to acyclovir and ganciclovir. Lancetii: 382.
20.
SalahuddinS.Z.AblashiD.V.MarkhamP.D.JosephsS.F.SturzeneggerS.KaplanM.HalliganG.BiberfeldP.Wong-StaalF.KramarskyB., and GalloR.C. (1986) Isolation of a new virus, HBLV, in patients with lymphoproliferative disorders. Science234: 596–601.
21.
SmeeD.F.MartinJ.C.VerheydenJ.P.H., and MatthewsT.R. (1983) Anti-herpesvirus activity of the acyclic nucleoside 9-(1,3-dihydroxy-2-propoxymethyl) guanine. Antimicrob Agents Chemother23: 676–682.
22.
SpearP.G., and RoizmanB. (1972) Proteins specified by herpes simplex virus. V. Purification and structural proteins of the herpesvirion. J Virol9: 143–159.
23.
TakahashiK.SonadaS.KawakamiK.MiyataK.OkiT.NagataT.OkunoT., and YamanishiK. (1988) Human herpesvirus 6 and Exanthem subitum. Lanceti: 1463.
24.
TedderR.S.BriggsM.CameronC.H.HonessR.RobertsonD., and WhittleH. (1987) A novel lymphotropic herpesvirus. Lancetii: 390–392.
25.
TowbinH.StaehelinT., and GordonJ. (1979) Electrophoretic transfer of proteins from polyacrylamide to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA76: 4350–4354.
26.
VromanB.LukaJ.RodriguezM., and PearsonG.R. (1985) Characterization of a major protein with a molecular weight of 160,000 associated with the viral capsid of Epstein–Barr virus. J Virol53: 107–113.
27.
WardK.N.GrayJ.J., and EfstathiouS. (1989) Primary human herpesvirus 6 infection in a patient following liver transplantation from a seropositive donor. J Med Virol28: 69–72.
28.
YamanishiK.OkunoT.ShirakiK.TakahashiM.KondoT.AsanoY., and KurataT. (1988) Identification of human herpesvirus-6 as a causal agent for Exanthem subitum. Lanceti: 1065–1067.
