Antigenic domain 1 of human cytomegalovirus glycoprotein B induces a multitude of different antibodies which, when combined, results in incomplete virus neutralization
Glycoprotein B (gB, gpUL55) is the major antigen for the induction of neutralizing antibodies against human cytomegalovirus (HCMV), making it an attractive molecule for active and passive immunoprophylaxis. The region between aa 552 and 635 of HCMV gB (termed AD-1) has been identified as the immunodominant target for the humoral immune response following natural infection. AD-1 represents a complex domain which requires a minimal continuous sequence of more than 70 aa for antibody binding. Neutralizing as well as non-neutralizing antibodies can bind to AD-1 in a competitive fashion. The fine specificity of AD-1-binding monoclonal antibodies (MAbs) and affinity-purified human polyclonal antibodies was analysed by using recombinant proteins containing single amino acid substitutions spanning the entire AD-1 domain. Our results revealed that all MAbs had individual patterns of binding to the mutant proteins indicating the presence of a considerable number of distinct antibody-binding sites on AD-1. The neutralization capacity of antibodies could not be predicted from their binding pattern to AD-1 mutant proteins. Polyclonal human antibodies purified from different convalescent sera showed identical binding patterns to the mutant proteins suggesting that the combined antibody specificities present in human sera are comparable between individuals. Neutralization capacities of polyclonal human AD-1 antibodies did not exceed 50% indicating that, during natural infection, a considerable proportion of non-neutralizing antibodies are induced and thus might provide an effective mechanism to evade complete virus neutralization.
AdlerS. P.,
StarrS. E.,
PlotkinS. A.,
HempflingS. H.,
BuisJ.,
ManningM. L.,
BestA. M.1995; Immunity induced by primary human cytomegalovirus infection protects against secondary infection among women of childbearing age. Journal of Infectious Diseases 171:26–32
BonciA.,
BracciL.,
CaudaiC.,
LozziL.,
MoschettiniD.,
NiccolaiN.,
ScarselliM.,
ValensinP. E.,
NeriP.1993; Characterization of immunoreactive octapeptides of human-cytomegalovirus gp58. European Journal of Biochemistry 215:383–387
BrittW. J.1984; Neutralizing antibodies detect a disulfide-linked glycoprotein complex within the envelope of human cytomegalovirus. Virology 135:369–378
BrittW. J.,
VuglerL. G.1992; Oligomerization of the human cytomegalovirus major envelope glycoprotein complex gB (gp55–116. Journal of Virology 66:6747–6754
BrittW. J.,
VuglerL.,
ButfiloskiE. J.,
StephensE. B.1990; Cell surface expression of human cytomegalovirus (HCMV) gp55–116 (gB): use of HCMV-recombinant vaccinia virus-infected cells in analysis of the human neutralizing antibody response. Journal of Virology 64:1079–1085
ChouS. W.,
DennisonK. M.1991; Analysis of interstrain variation in cytomegalovirus glycoprotein B sequences encoding neutralization-related epitopes. Journal of Infectious Diseases 163:1229–1234
DarlingtonJ.,
SuperM.,
PatelK.,
GrundyJ. E.,
GriffithsP. D.,
EmeryV. C.1991; Use of the polymerase chain reaction to analyse sequence variation within a major neutralizing epitope of glycoprotein B (gp58) in clinical isolates of human cytomegalovirus. Journal of General Virology 72:1985–1989
FowlerK. B.,
StagnoS.,
PassR. F.,
BrittW. J.,
BollT. J.,
AlfordC. A.1992; The outcome of congenital cytomegalovirus infection in relation to maternal antibody status. see comments New England Journal of Medicine 326:663–667
GonczolE.,
deTaisneC.,
HirkaG.,
BerencsiK.,
LinW. C.,
PaolettiE.,
PlotkinS.1991; High expression of human cytomegalovirus (HCMV)-gB protein in cells infected with a vaccinia-gB recombinant: the importance of the gB protein in HCMV immunity. Vaccine 9:631–637
JonjicS.,
PavicI.,
PolicB.,
CrnkovicI.,
LucinP.,
KoszinowskiU. H.1994; Antibodies are not essential for the resolution of primary cytomegalovirus infection but limit dissemination of recurrent virus. Journal of Experimental Medicine 179:1713–1717
KariB.,
GehrzR.1991; Biochemical and immunological analysis of discontinuous epitopes in the family of human cytomegalovirus glycoprotein complexes designated gC-I. Journal of General Virology 72:1975–1983
KariB.,
LiuY. N.,
GoertzR.,
LussenhopN.,
StinskiM. F.,
GehrzR.1990; Structure and composition of a family of human cytomegalovirus glycoprotein complexes designated gC-I (gB. Journal of General Virology 71:2673–2680
KleinM.,
SchoppelK.,
AmvrossiadisN.,
MachM.1999; Strain-specific neutralization of human cytomegalovirus isolates by human sera. Journal of Virology 73:878–886
KniessN.,
MachM.,
FayJ.,
BrittW. J.1991; Distribution of linear antigenic sites on glycoprotein gp55 of human cytomegalovirus. Journal of Virology 65:138–146
KropffB.,
MachM.1997; Identification of the gene coding for rhesus cytomegalovirus glycoprotein B and immunological analysis of the protein. Journal of General Virology 78:1999–2007
KropffB.,
LandiniM. P.,
MachM.1993; An ELISA using recombinant proteins for the detection of neutralizing antibodies against human cytomegalovirus. Journal of Medical Virology 39:187–195
MarshallG. S.,
RabalaisG. P.,
StoutG. G.,
WaldeyerS. L.1992; Antibodies to recombinant-derived glycoprotein B after natural human cytomegalovirus infection correlate with neutralizing activity. Journal of Infectious Diseases 165:381–384
MeyerH.,
MasuhoY.,
MachM.1990; The gp116 of the gp58/116 complex of human cytomegalovirus represents the amino-terminal part of the precursor molecule and contains a neutralizing epitope. Journal of General Virology 71:2443–2450
MeyerH.,
SundqvistV.-A.,
PereiraL.,
MachM.1992; Glycoprotein gp116 of human cytomegalovirus contains epitopes for strain-common and strain-specific antibodies. Journal of General Virology 73:2375–2383
MyersR. M.1989; Region-specific mutagenesis. In Current Protocols in Molecular Biology , unit 8.3 Edited by
AusubelF. M.,
BrentR.,
KingstonR. E.,
MooreD. D.,
SeidmanJ. G.,
StruhlK.
New York: John Wiley;
OhlinM.,
SundqvistV. A.,
MachM.,
WahrenB.,
BorrebaeckC. A.1993; Fine specificity of the human immune response to the major neutralization epitopes expressed on cytomegalovirus gp58/116 (gB), as determined with human monoclonal antibodies. Journal of Virology 67:703–710
QadriI.,
NavarroD.,
PazP.,
PereiraL.1992; Assembly of conformation-dependent neutralizing domains on glycoprotein B of human cytomegalovirus. Journal of General Virology 73:2913–2921
RappM.,
MesserleM.,
LucinP.,
KoszinowskiU. H.1993; In vivo protection studies with MCMV glycoproteins gB and gH expressed by vaccinia virus. In Multidisciplinary Approach to Understanding Cytomegalovirus Disease pp 327–332 Edited by
MichelsonS.,
PlotkinS. A.
Amsterdam: Excerpta Medica;
ReddehaseM. J.,
BalthesenM.,
RappM.,
JonjicS.,
PavicI.,
KoszinowskiU. H.1994; The conditions of primary infection define the load of latent viral genome in organs and the risk of recurrent cytomegalovirus disease. Journal of Experimental Medicine 179:185–193
RoyD. M.,
GrundyJ. E.,
EmeryV. C.1993; Sequence variation within neutralizing epitopes of the envelope glycoprotein B of human cytomegalovirus: comparison of isolates from renal transplant recipients and AIDS patients. Journal of General Virology 74:2499–2505
SchoppelK.,
HassfurtherE.,
BrittW. J.,
OhlinM.,
BorrebaeckC. A.,
MachM.1996; Antibodies specific for the antigenic domain 1 (AD-1) of glycoprotein B (gpUL55) of human cytomegalovirus bind to different substructures. Virology 216:133–145
SchoppelK.,
KropffB.,
SchmidtC.,
VornhagenR.,
MachM.1997; The humoral immune response against human cytomegalovirus is characterized by a delayed synthesis of glycoprotein specific antibodies. Journal of Infectious Diseases 175:533–544
SpindlerK. R.,
RosserD. S.,
BerkA. J.1984; Analysis of adenovirus transforming proteins from early regions 1A and 1B with antisera to inducible fusion antigens produced in Escherichia coli. Journal of Virology 49:132–141
TanakaA.,
MoriuchiH.,
HirotaK.,
NumazakiY.1991; Neutralizing antibody response to cytomegalovirus in seropositive pregnant women. Journal of Medical Virology 34:85–88
UrbanM.,
BrittW.,
MachM.1992; The dominant linear neutralizing antibody-binding site of glycoprotein gp86 of human cytomegalovirus is strain specific. Journal of Virology 66:1303–1311
UrbanM.,
WinklerT.,
LandiniM. P.,
BrittW.,
MachM.1994; Epitope-specific distribution of IgG subclasses against antigenic domains on glycoproteins of human cytomegalovirus. Journal of Infectious Diseases 169:83–90
UrbanM.,
KleinM.,
BrittW. J.,
HassfurtherE.,
MachM.1996; Glycoprotein H of human cytomegalovirus is a major antigen for the neutralizing humoral immune response. Journal of General Virology 77:1537–1547
UtzU.,
BrittW.,
VuglerL.,
MachM.1989; Identification of a neutralizing epitope on glycoprotein gp58 of human cytomegalovirus. Journal of Virology 63:1995–2001
WagnerB.,
KropffB.,
KalbacherH.,
BrittW.,
SundqvistV. A.,
OstbergL.,
MachM.1992; A continuous sequence of more than 70 amino acids is essential for antibody binding to the dominant antigenic site of glycoprotein gp58 of human cytomegalovirus. Journal of Virology 66:5290–5297
WangJ. B.,
AdlerS. P.,
HempflingS.,
BurkeR. L.,
DuliegeA. M.,
StarrS. E.,
PlotkinS. A.1996; Mucosal antibodies to human cytomegalovirus glycoprotein B occur following both natural infection and immunization with human cytomegalovirus vaccines. Journal of Infectious Diseases 174:387–392
WernerB. G.,
SnydmanD. R.,
FreemanR.,
RohrerR.,
TilneyN. L.,
KirkmanR. L.1993; Cytomegalovirus immune globulin for the prevention of primary CMV disease in renal transplant patients: analysis of usage under treatment IND status. The Treatment IND Study Group. Transplantation Proceedings 25:1441–1443
Antigenic domain 1 of human cytomegalovirus glycoprotein B induces a multitude of different antibodies which, when combined, results in incomplete virus neutralization