The attachment glycoprotein G plays a major role in the antigenic variability of respiratory syncytial (RS) virus. We have expressed from recombinant baculoviruses antigenic group A and group B RS virus G proteins (designated bacAG for the group A and bacBG for the group B virus G protein). The insect cell-produced G proteins migrated more rapidly in SDS-PAGE as compared to HEp-2 cell-derived G proteins owing to glycosylation differences. Antigenicity was tested by immunofluorescence; five of five group cross-reactive, five of six group A-specific, and six of six group B-specific MAbs reacted appropriately with bacAG and/or bacBG. In addition, bacAG and bacBG reacted with human polyclonal antibodies to RS virus. Cotton rats were immunized with bacAG, bacBG or a control lysate and challenged intranasally with a group A RS virus. The bacAG-immunized group had a statistically significant reduction in viral replication in the lungs (lung titres as mean log10 p.f.u./g ± sd, bacAG = 3.1 ± 1.2; control = 4.8 ± 0.6, P = 0.013). The bacBG-immunized group showed less reduction in viral titres (bacBG lung titres = 4.1 ± 0.6, P = 0.13 for bacBG compared to control). Thus, as expected, homologous protein (bacAG) immunization provided more protection against viral replication than immunization with the heterologous protein (bacBG). The G protein of RS virus expressed in insect cells had antigenic and immunogenic features which were similar to that of the G protein expressed in mammalian cells. The baculovirus-expressed G proteins should be useful for the study of immune responses to RS viruses.
AndersonL. J.,
BinghamP.,
HierholzerJ. C.1988; Neutralization of respiratory syncytial virus by individual and mixtures of F and G protein monoclonal antibodies. Journal of Virology 62:4232–4238
BallL. A.,
YoungK. K. Y.,
AndersonK.,
CollinsP. L.,
WertzG. W.1986; Expression of the major glycoprotein G of human respiratory syncytial virus from recombinant vaccinia virus vectors. Proceedings of the National Academy of Sciences, USA 83:246–250
CollinsP. L.,
MottetG.1992; Oligomerization and post–translational processing of glycoprotein G of human respiratory syncytial virus: altered O-glycosylation in the presence of brefeldin A. Journal of General Virology 73:849–863
CollinsP. L.,
HuangY. T.,
WertzG. W.1984; Identification of a tenth mRNA of respiratory syncytial virus and assignment of polypeptides to the 10 viral genes. Journal of Virology 49:572–578
García-BarrenoB.,
PalomoC.,
PenasC.,
DelgadoT.,
Perez-BrenaP.,
MeleroJ. A.1989; Marked differences in the antigenic structure of human respiratory syncytial virus F and G glycoproteins. Journal of Virology 63:925–932
GlezenW. P.,
ParedesA.,
AllisonJ. E.,
TaberL. H.,
FrankA.L.1981; Risk of respiratory syncytial virus infection for infants from low-income families in relationship to age, sex, ethnic group, and maternal antibody level. Journal of Pediatrics 98:708–715
GruberC.,
LevineS.1985; Respiratory syncytial virus polypeptides. IV. The oligosaccharides of the glycoproteins. Journal of General Virology 66:417–432
HendersonF. W.,
CollierA. M.,
ClydeW. A.,
DennyF. W.1979; Respiratory syncytial virus infections, reinfections and immunity. New England Journal of Medicine 300:530–534
JarvisD. L.,
SummersM. D.1992; Baculovirus expression vectors. In Recombinant DNA Vaccines: Rationale and Strategy pp 265–291 Edited by
IsaacsonR. E.
New York: Marcel Dekker;
JohnsonP. R.,
OlmstedR. A.,
PrinceG. A.,
MurphyB. R.,
AllingD. W.,
WalshE. E.,
CollinsP. L.1987a; Antigenic relatedness between glycoproteins of human respiratory syncytial virus subgroups A and B: evaluation of the contributions of F and G glycoproteins to immunity. Journal of Virology 61:3163–3166
JohnsonP. R.,
SpriggsM. K.,
OlmstedR. A.,
CollinsP. L.1987b; The G glycoprotein of human respiratory syncytial viruses of subgroups A and B: extensive sequence divergence between antigenically related proteins. Proceedings of the National Academy of Sciences, USA 84:5625–5629
Martin-GallardoA.,
FleischerE.,
DoyleS. A.,
ArumughamR.,
CollinsP. L.,
HildrethS. W.,
ParadisoP. R.1993; Expression of the G glycoprotein gene of human respiratory syncytial virus in Salmonella typhimurhim
. Journal of General Virology 74:453–458
MufsonM. A.,
BelsheR. B.,
örvellC.,
NorrbyE.1987; Subgroup characteristics of respiratory syncytial virus strains recovered from children with two consecutive infections. Journal of Clinical Microbiology 25:1536–1539
NorrbyE.,
MufsonM. A.,
AlexanderH.,
HoughtenR. A.,
LernerR. A.1987; Site-directed serology with synthetic peptides representing the large glycoprotein G of respiratory syncytial virus. Proceedings of the National Academy of Sciences, USA 84:6572–6576
OlmstedR. A.,
PrinceG. A.,
MurphyB. R.,
JohnsonP. R.,
MossB.,
ChanockR. M.,
CollinsP. L.1986; Expression of the F glycoprotein of respiratory syncytial virus by a recombinant vaccinia virus: comparison of the individual contributions of the F and G glycoproteins to host immunity. Proceedings of the National Academy of Sciences, USA 83:7462–7466
PalomoC.,
García-BarrenoB.,
PeñasC.,
MeleroJ. A.1991; The G protein of human respiratory syncytial virus: significance of carbohydrate side-chains and the C-terminal end to its antigenicity. Journal of General Virology 72:669–675
RuedaP.,
Garci’a-BarrenoB.,
MeleroJ. A.1994; Loss of conserved cysteine residues in the attachment (G). Glycoprotein of two human respiratory syncytial virus escape mutants that contain multiple A → G substitutions (hypermutations). Virology 198:653–662
StottE. J.,
BallL. A.,
YoungK. K.,
FurzeJ.,
WertzG. W.1986; Human respiratory syncytial virus glycoprotein G expressed from a recombinant vaccinia virus vector protects mice against live-virus challenge. Journal of Virology 60:607–613
StottE. J.,
TaylorG.,
BallL. A.,
AndersonK.,
YoungK. Y.,
KingA. M. Q.,
WertzG. W.1987; Immune and histopathological responses in animals vaccinated with recombinant vaccinia viruses that express individual genes of human respiratory syncytial virus. Journal of Virology 61:3855–3861
SullenderW. M.,
WertzG. W.1991; The unusual attachment glycoprotein of the respiratory syncytial viruses: structure, maturation, and role in immunity. In The Paramyxoviruses pp 383–406 Edited by
KingsburyD.
New York: Plenum Press;
SullenderW. M.,
AndersonK.,
WertzG. W.1990; The respiratory syncytial virus subgroup B attachment glycoprotein: analysis of sequence, expression from a recombinant vector, and evaluation as an immunogen against homologous and heterologous subgroup virus challenge. Virology 178:195–203
WalshE. E.,
HallC. B.,
BriselliM.,
BrandrissM. W.,
SchlesingerJ. J.1987; Immunization with glycoprotein subunits of respiratory syncytial virus to protect cotton rats against viral infection. Journal of Infectious Diseases 155:1198–1204
WalshE. E.,
HallC. B.,
SchlesingerJ. J.,
BrandrissM. W.,
HildrethS.,
ParadisoP.1989; Comparison of antigenic sites of subtype-specific respiratory syncytial virus attachment proteins. Journal of General Virology 70:2953–2961
WarisM.1991; Pattern of respiratory syncytial virus epidemics in Finland: two-year cycles with alternating prevalence of groups A and B. Journal of Infectious Diseases 163:464–469
WathenM. W.,
BrideauR. J.,
ThomsenD. R.,
MurphyB. R.1989; Characterization of a novel human respiratory syncytial virus chimeric FG glycoprotein expressed using a baculovirus vector. Journal of General Virology 70:2625–2635
WathenM. W.,
AeedP. A.,
ElhammerA. P.1991; Characterization of oligosaccharide structures on a chimeric respiratory syncytial virus protein expressed in insect cell line Sf9. Biochemistry 30:2863–2868
WellsD. E.,
VuglerL. G.,
BrittW.1990; Structural and immunological characterization of human cytomegalovirus gp55–116 (gB) expressed in insect cells. Journal of General Virology 71:873–880
WertzG. W.,
CollinsP. L.,
HuangY.,
GruberC.,
LevineS.,
BallL. A.1985; Nucleotide sequence of the G protein gene of human respiratory syncytial virus reveals an unusual type of viral membrane protein. Proceedings of the National Academy of Sciences, USA 82:4075–7049
WertzG. W.,
KriegerM.,
BallL. A.1989; Structure and cell surface maturation of the attachment glycoprotein of human respiratory syncytial virus in a cell line deficient in O glycosylation. Journal of Virology 63:4767–4776
WickhamT. J.,
DavisT.,
GranadosR. R.,
ShulerM. L.,
WoodH. A.1992; Screening of insect cell lines for the production of recombinant proteins and infectious virus in the baculovirus expression system. Biotechnology 8:391–396