Cross-reactive neutralization epitopes on VP4 of human rotavirus (HRV) were analysed by the use of VP4-specific neutralizing monoclonal antibodies (N-MAbs) and MAb-resistant mutants. Seven anti-VP4 N-MAbs obtained in this study by using HRV serotypes 1 and 3 as immunizing antigens showed a variety of cross-reactivity patterns to 20 HRV strains with different serotype specificity in neutralization tests and a broader cross-reactivity to them was found for four N-MAbs in an enzyme-linked immunosorbent assay. On the basis of the reactivity patterns against rotaviruses in neutralization tests, these seven N-MAbs were classified into four groups. Cross-neutralization tests using a total of 12 pairs of MAbs and resistant mutants, including five pairs which had been prepared previously, showed that VP4 of HRV (strain KU) contained two independent antigenic regions. One, region C1, was recognized by a single MAb (YO-2C2) and the other was made up of two antigenic regions (C2 and C3) which overlapped operationally. Identification of amino acid substitution sites on VP4 of representative mutants of HRV strain KU indicated that amino acid positions 385 or 392 and 428 or 433 were critical for the C2 and C3 regions, respectively. These results suggested that regions C2 and C3 exist as conformational antigenic sites.
BurnsJ. W., GreenbergH. B., ShawR. D., EstesM. K.1988; Functional and topographical analyses of epitopes on the haemagg-lutinin (VP4) of the simian rotavirus SA11. Journal of Virology 62:2164–2172
CoulsonB. S., TursiJ. M., McadamW. J., BishopR. F.1986; Derivation of neutralizing monoclonal antibodies to human rotaviruses and evidence that an immunodominant neutralization site is shared between serotypes 1 and 3. Virology 154:301–312
Dyall-SmithM. L., LazdinsI., TregearG. W., HolmesI. H.1986; Location of the major antigenic sites involved in rotavirus serotype-specific neutralization. Proceedings of the National Academy of Sciences, U.S.A 83:3465–3468
FloresJ., MidthunK., HoshinoY., GreenK., GorzigliaM., KapikianA. Z., ChanockR. M.1986; Conservation of the fourth gene among rotaviruses recovered from asymptomatic newborn infants and its possible role in attenuation. Journal of Virology 60:972–979
GorzigliaM., HoshinoY., Buckler-WhiteA., BlumentalsI., GlassR., FloresJ., KapikianA. Z., ChanockR. M.1986; Conservation of amino acid sequence of VP8 and cleavage region of 84-kDa outer capsid protein among rotaviruses recovered from asymptomatic neonatal infection. Proceedings of the National Academy of Sciences, U.S.A 83:7039–7043
GorzigliaM., GreenK. Y., NishikawaK., TaniguchiK., JonesR., KapikianA. Z., ChanockR. M.1988; Sequence of the fourth gene of human rotaviruses recovered from asymptomatic or symptomatic infections. Journal of Virology 62:2978–2984
GreenbergH. B., KalicaA. R., WyattR. G., JonesR. W., KapikianA. Z., ChanockR. M.1981; Rescue of noncultiva-table human rotavirus by gene reassortment during mixed infection with ts mutants of cultivatable bovine rotavirus. Proceedings of the National Academy of Sciences, U.S.A 78:420–424
GreenbergH. B., FloresJ., KalicaA. R., WyattR. G., JonesR.1983; Gene coding assignments for growth restriction, neutralization and subgroup specificities of the W and DS-1 strains of human rotavirus. Journal of General Virology 64:313–320
KalicaA. R., GreenbergH. B., WyattR. G., FloresJ., SeneroM. M., KapikianA. Z., ChanockR. M.1981; Genes of human (strain Wa) and bovine (strain UK) rotaviruses that code for neutralization and subgroup antigens. Virology 112:385–390
KalicaA. R., FloresJ., GreenbergH. B.1983; Identification of the rotaviral gene that codes for the hemagglutinin and protease-enhanced plaque formation. Virology 125:194–205
KaljotK. T., ShawR. D., RubinD. H., GreenbergH. B.1988; Infectious rotavirus enters cells by direct cell membrane penetration, not by endocytosis. Journal of Virology 62:1136–1144
KapikianA. Z., ChanockR. M.1985; Rotaviruses. In Virology pp. 863–906FieldsB. N., KnipeD. M., ChanockR. M., MelnickJ. L., RoizmanB., ShopeR. E. Edited by New York: Raven Press;
KapikianA. Z., FloresJ., HoshinoY., GlassR.L, GorzigliaM., ChanockR. M.1986; Rotavirus: the major etiologic agent of severe infantile diarrhea may be controllable by a ‘Jennerian’ approach to vaccination. Journal of Infectious Diseases 153:815–822
KitaokaS., FukuharaN., TazawaF., SuzukiH., SatoT., KonnoT., EbinaT., IshidaN.1986; Characterization of monoclonal antibodies against human rotavirus hemagglutinin. Journal of Medical Virology 19:313–323
MackowE. R., ShawR. D., MatsuiS. M., VoP. T., BenfieldD. A., GreenbergH. B.1988a; Characterization of homotypic and heterotypic VP7 neutralization sites of rhesus rotavirus. Virology 165:511–517
MackowE. R., ShawR. D., MatsuiS. M., VoP. T., DangM.-N., GreenbergH. B.1988b; The rhesus rotavirus gene encoding protein VP3: location of amino acids involved in homologous and heterologous rotavirus neutralization and identification of a putative fusion region. Proceedings of the National Academy ofSciences, U.S.A 85:645–649
MoritaY., TaniguchiK., UrasawaT., UrasawaS.1988; Analysis of serotype-specific neutralization epitopes on VP7 of human rotavirus by the use of neutralizing monoclonal antibodies and antigenic variants. Journal of General Virology 69:451–458
NishikawaK., TaniguchiK., TorresA., HoshinoY., GreenK. Y., KapikianA. Z., ChanockR. M., GorzigliaM.1988; Comparative analysis of the VP3 gene of divergent strains of the rotaviruses simian SA11 and bovine Nebraska calf diarrhea virus. Journal of Virology 62:4022–4026
OffitP. A., ClarkH. F., StroopW. G., TwistE. M., PlotkinS. A.1983; The cultivation of human rotavirus, strain ‘Wa,’ to high titer in cell culture and characterization of the viral structural polypeptides. Journal of Virological Methods 7:29–40
OffitP. A., ShawR. D., GreenbergH. B.1986; Passive protection against rotavirus-induced diarrhea by monoclonal antibodies to surface proteins vp3 and vp7. Journal of Virology 58:700–703
StockerJ. W., ForsterH. K., MiggianoV., StähliC., StaigerG., TakacsB., StaehelinT.1982; Generation of 2 new mouse myeloma cell lines ‘PAI’ and ‘PAI-O’ for hybridoma production. Research Disclosure 217:155–157
TaniguchiK., UrasawaS., UrasawaT.1983; Monoclonal antibodies reactive with different sites of N and H antigenic particles of poliovirus. Journal of General Virology 64:2803–2808
TaniguchiK., UrasawaS., UrasawaT.1985; Preparation and characterization of neutralizing monoclonal antibodies with different reactivity patterns to human rotaviruses. Journal of General Virology 66:1045–1053
TaniguchiK., MoritaY., UrasawaT., UrasawaS.1987; Cross-reactive neutralization epitopes on VP3 of human rotavirus: analysis with monoclonal antibodies and antigenic variants. Journal of Virology 61:1726–1730
TaniguchiK., HoshinoY., NishikawaK., GreenK. Y., MaloyW. L., MoritaY., UrasawaS., KapikianA. Z., ChanockR. M., GorzigliaM.1988a; Cross-reactive and serotype-specific neutralization epitopes on VP7 of human rotavirus: nucleotide sequence analysis of antigenic mutants selected with monoclonal antibodies. Journal of Virology 62:1870–1874
TaniguchiK., MaloyW. L., NishikawaK., GreenK. Y., HoshinoY., UrasawaS., KapikianA. Z., ChanockR. M., GorzigliaM.1988b; Identification of cross-reactive and serotype 2-specific neutralization epitopes on VP3 of human rotavirus. Journal of Virology 62:2421–2426
TaniguchiK., NishikawaK., UrasawaT., UrasawaS., MidthunK., KapikianA. Z., GorzigliaM.1989; Complete nucleotide sequence of the gene encoding VP4 of a human rotavirus (strain K8) which has unique VP4 neutralization epitopes. Journal of Virology 63:4101–4106
UrasawaS., UrasawaT., TaniguchiK.1986; Genetic reassortment between two human rotaviruses having different serotype and subgroup specificities. Journal of General Virology 67:1551–1559
UrasawaS., UrasawaT., TaniguchiK., WakasugiF., KobayashiN., ChibaS., SakuradaN., MoritaM., MoritaO., TokiedaM., KawamotoH., MinekawaY., OhsetoM.1989; Survey of human rotavirus serotypes in different locales in Japan by enzyme-linked immunosorbent assay with monoclonal antibodies. Journal of Infectious Diseases 160:44–51
UrasawaT., UrasawaS., TaniguchiK., KobayashiN.1988; Preparation of a human rotavirus reassortant with dual serotype specificity, VP3 of serotype 4 and VP7 of serotype 3. Sapporo Medical Journal 57:373–378
VesikariT., IsolauriE., D’HondtE., DelemA., AndréF. E., ZissisG.1984; Protection of infants against rotavirus diarrhoea by RIT 4237 attenuated bovine rotavirus strain vaccine. Lancet i:977–981
WardR. L., KnowltonD. R., SchiffG. M., HoshinoY., GreenbergH. B.1988; Relative concentrations of serum neutralizing antibody to VP3 and VP7 proteins in adults infected with a human rotavirus. Journal of Virology 62:1543–1549
WyattR. G., GreenbergH. B., JamesW. D., PittmanA. L., KalicaA. R., FloresJ., ChanockR. M., KapikianA. Z.1982; Definition of human rotavirus serotypes by plaque reduction assay. Infection and Immunity 37:110–115
YoshinoK., TaniguchiS.1965; Studies on the neutralization of herpes simplex virus. III. Mechanism of the antibody-potentiating action of complement. Virology 26:61–72