The major capsid protein (MCP) of human cytomegalovirus (HCMV) was expressed in three portions as β-galactosidase fusion proteins, covering about 75 % of the open reading frame (ORF). Fusion protein SH 1 contained nucleotides 101 to 1243 of the ORF, fusion protein FS 1 contained nucleotides 1944 to 3089 and fusion protein SS 1 covered nucleotides 2624 to 3793. The recombinant proteins were tested for their immunoreactivity with human sera. Fusion protein FS 1 was found to represent the immunodominant region. The recombinant proteins were used to generate polyvalent rabbit antisera to investigate cross-reactivities with the major capsid protein (VP5) of herpes simplex virus type 1 (HSV-1). A monospecific antiserum raised against the fusion protein close to the N terminus of the MCP, as well as a monoclonal antibody and a monospecific rabbit antiserum directed against the viral MCP, cross-reacted with the VP5 as shown by immunoblotting and immunofluorescence. In order to detect common epitopes of the major capsid proteins of HCMV and HSV-1, the recombinant proteins were conjugated to CNBr-activated Sepharose and taken for purification of MCP antibodies from HCMV and HSV-1 seropositive individuals. Using this affinity chromatography method, cross-reactivity could be observed with HCMV- and HSV-positive human antisera in immunoblot experiments.
BalachandranN.,
ObaD. E.,
Hutt-FletcherL. M.1987; Antigenic cross-reactions among herpes simplex virus types 1 and 2, Epstein Barr virus and cytomegalovirus. Journal of Virology 61:1125–1135
BernsteinD. I.,
GarrattyE.,
LovettM. A.,
BrysonY. J.1985; Comparison of Western blot analysis to micro neutralization for the detection of type-specific herpes simplex virus antibodies. Journal of Medical Virology 15:223–230
Bibor-HardyV.,
DagenaisA.,
SimardR.1985; In situ localization of the major capsid protein during lytic infection by herpes simplex virus. Journal of General Virology 66:897–901
BrökerM.1986; Vectors for regulated high-level expression of proteins fused to truncated forms of Escherichia coliβ-galactosidase. Gene Analysis Techniques 3:53–57
CheeM.,
RudolphS.-A.,
PlachterB.,
BarrellB.,
JahnG.1989; Identification of the major capsid protein gene of human cytomegalovirus. Journal of Virology 63:1345–1353
DubeyL.,
SteinbergS. P.,
LarussaP.,
OhP.,
GershonA. A.1988; Western blot analysis of antibody to varicella-zoster virus. Journal of Infectious Diseases 157:882–888
EberleR.,
CourtneyJ.1981; Assay for type-specific and type-common antibodies to herpes simplex types 1 and 2 in human sera. Infection and Immunity 31:1062–1070
EberleR.,
MouS.-W.1983; Relative titres of antibodies to individual polypeptide antigens of herpes simplex virus type 1 in human sera. Journal of Infectious Diseases 148:436–444
EberleR.,
MouS.-W.,
ZaiaJ. A.1985; The immune response to herpes simplex virus: comparison of the specificity and relative titres of serum antibodies directed against viral polypeptides following primary herpes simplex virus type 1 infections. Journal of Medical Virology 16:147–162
EdsonC. M.,
HoslerB. A.,
RespessR. A.,
WatersD. J.,
Thorley-LawsonD.1985; Cross-reactivity between herpes simplex virus glycoprotein B and a 63,000-dalton varicella-zoster virus envelope glycoprotein. Journal of Virology 56:333–336
EllingerS.,
GlockshuberR.,
JahnG.,
PlückthunA.1989; Cleavage of procaryotically expressed human immunodeficiency virus fusion proteins by factor Xa and application in Western blot (immunoblot) assays. Journal of Clinical Microbiology 27:971–976
HonessR. W.,
PowellK. L.,
RobinsonD. J.,
SimC.,
WatsonD. H.1974; Type specific and type common antigens in cells infected with herpes simplex type 1 and on the surfaces of naked and enveloped particles of the virus. Journal of General Virology 22:159–169
IrmiereA.,
GibsonW.1983; Isolation and characterization of a non-infectious virion-like particle released from cells infected with human strains of cytomegalovirus. Virology 130:118–133
JahnG.,
SchollB. C.,
TraupeB.,
FleckensteinB.1987a; The two major structural phosphoproteins (pp65 and ppl50) of human cytomegalovirus and their antigenic properties. Journal of General Virology 68:1327–1337
JahnG.,
KouzaridesT.,
MachM.,
SchollB.-C.,
PlachterB.,
TraupeB.,
PreddieE.,
SatchwellS. C.,
FleckensteinB.,
BarrellB. G.1987b; Map position and nucleotide sequence of the gene for the large structural phosphoprotein of human cytomegalovirus. Journal of Virology 61:1358–1367
KahlonJ.,
LakemanF. D.,
AckermannM.,
WhitleyR. J.1986; Human antibody response to herpes simplex virus-specific polypeptides after primary and recurrent infection. Journal of Clinical Microbiology 23:725–730
KillingtonR. A.,
YeoJ.,
HonessR. W.,
WatsonD. H.,
DuncanB. E.,
HalliburtonI. W.,
MumfordJ.1977; Comparative analyses of the proteins and antigens of five herpesviruses. Journal of General Virology 37:297–310
KühnJ. E.,
DunklerG.,
MunkK.,
BraunR. W.1987; Analysis of the IgM and IgG antibody response against herpes simplex virus type 1 (HSV-1) structural and non-structural proteins. Journal of Medical Virology 23:135–150
LandiniM. P.,
ReM. C.,
MiroloG.,
BaldassarriB.,
La PlacaM.1985; Human immune response to cytomegalovirus structural polypeptides studied by immunoblotting. Journal of Medical Virology 17:303–311
LawrenceG. L.,
CheeM.,
CraxtonM. A.,
GompelsU. A.,
HonessR. W.,
BarrellB. G.1990; Human herpesvirus 6 is closely related to human cytomegalovirus. Journal of Virology 64:287–299
LittlerE.,
HalliburtonI. W.,
PowellK. L.,
SnowdenB. W.,
ArrandJ. R.1988; Immunological conservation between Epstein-Barr virus and herpes simplex virus. Journal of General Virology 69:2021–2031
ManningA.,
BuchanA.,
SkinnerG. R. B.,
DurhamJ.,
ThompsonH.1988; The immunological relationship between canine herpesvirus and four other herpesviruses. Journal of General Virology 69:1601–1608
MerrilC. R.,
GoldmanD.,
SedmanS. A.,
EbertM. H.1981; Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. Science 211:1437–1438
MessingJ.,
GronenbornB.,
Müller-HillB.,
HofschneiderP. H.1977; Filamentous coliphage M13 as a cloning vehicle: insertion of a HindII fragment of the lac regulatory region in Ml3 replicative form in vitro
. Proceedings of the National Academy of Sciences, U.S.A 74:3642–3646
MiddeldorpJ. M.,
MeloenR. H.1988; Epitope-mapping on the Epstein-Barr virus major capsid protein using systematic synthesis of overlapping oligopeptides. Journal of Virological Methods 21:147–159
PereiraL.,
HoffmanM.,
CremerN.1982; Electrophoretic analysis of polypeptides immune-precipitated from cytomegalovirus-infected cell extracts by human sera. Infection and Immunity 36:933–942
RossL. J. N.,
FrazierJ. A.,
BiggsP. M.1972; An antigen common to some avian and mammalian herpesviruses. In Oncogenesis and Herpesviruses pp. 480–484BiggsP. M.
Edited by Lyon: IARC;
SchollB.-C.,
von HintzensternJ.,
BorischB.,
TraupeB.,
BrökerM.,
JahnG.1988; Prokaryotic expression of immunogenic polypeptides of the large phosphoprotein (pp 150) of human cytomegalovirus. Journal of General Virology 69:1195–1204
ShirakiK.,
OkunoT.,
YamanishiK.,
TakahashiM.1982; Polypeptides of varicella-zoster virus (VZV) and immunological relationship of VZV and herpes simplex virus (HSV). Journal of General Virology 61:255–269
SnowdenB. W.,
HalliburtonI. W.1985; Identification of crossreacting glycoproteins of four herpesviruses by Western blotting. Journal of General Virology 66:2039–2044
SpearP. G.,
RoizmanB.1972; Proteins specified by herpes simplex virus V. Purification of structural proteins of the herpesvirion. Journal of Virology 9:143–159
TowbinH.,
StaehelinT.,
GordonJ.1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences, U.S.A 76:4350–4354
VromanB.,
LukaJ.,
RodriguezM.,
PearsonG. R.1985; Characterization of a major protein with a molecular weight of 160000 associated with the viral capsid of Epstein-Barr virus. Journal of Virology 53:107–113
WolfH.,
MotzM.,
KühbedeR.,
JilgW.,
FanJ.,
PiG. H.,
ZengY.1987; Development of a set of EBV-specific antigens with recombinant gene technology for diagnosis of EBV-related malig-nant and non-malignant diseases. In Epstein-Barr Virus and Human Disease pp. 179–182LevineP. H.
Edited by Clifton, New Jersey: The Humana Press;
YamauchiM.,
NishiyamaY.,
FujiokaH.,
IsomuraS.,
MaenoK.1985; On the intracellular transport and the nuclear association of human cytomegalovirus structural proteins. Journal of General Virology 66:675–684