Cell lines of primate origin carry receptors on their plasma membrane which are responsible for the specific binding of poliovirus. This paper describes the isolation and characterization of a monoclonal antibody reacting with the plasma membrane of HeLa cells. The antibody (D171) was selected for its protection of HeLa cells against the cytopathic effect of poliovirus type 1. This protection was found to extend to all three viral serotypes, while the replication of five other viruses in HeLa cells was not affected. The 125I-labelled purified antibody did not react with cell lines derived from pig, dog or rodents but bound specifically to all lines of human or primate origin. Immunoglobulin or Fab fragments of D171 prevented the binding of 35S-labelled poliovirus to HeLa cells. Conversely, nearly all binding sites of 125I-labelled D171 immunoglobulins or Fab fragments could be blocked after preincubation of HeLa cells with poliovirus. These results indicate that D171 recognizes the poliovirus receptor site on different susceptible cells and that practically all D171 binding sites are involved in the specific attachment of poliovirus to the plasma membrane. To determine whether the epitope recognized by D171 could be separated from the receptor for poliovirus, human-mouse cell hybrids were prepared and analysed. In all 40 clones tested, the susceptibility to poliovirus correlated with the binding of D171.
ChildsR. A.,
GregoriouM.,
ScudderP.,
ThorpeS. J.,
ReesA. R.,
FeiziT.1984; Blood group-active carbohydrate chains on the receptor for epidermal growth factor of A431 cells. EMBO Journal 3:2227–2233
CrowellR. L.,
LandauB. J.1983; Receptors in the initiation of picornavirus infections. In Comprehensive Virology vol 18: pp 1–42 Edited by
H.Fraenkel-Conrat,
WagnerR. R.
New York & London: Plenum Press;
CrowellR. L.,
SiaKJ.-S.1978; Receptor for group B coxsackieviruses: characterization and extraction from HeLa cell plasma membranes. In Perspectives in Virology vol 10: pp 39–55 Edited by
PollardM.
New York: Raven Press;
EminiE. A.,
JamesonB. A.,
WimmerE.1983; Priming for and induction of anti-poliovirus neutralizing antibodies by synthetic peptides. Nature, London 304:699–703
EyP. L.,
ProwseS. J.,
JenkinC. R.1978; Isolation of pure IgG1, IgG2a and IgG2b immunoglobulins from mouse serum using Protein A-Sepharose. Immunochemistry 15:429–436
FingerothJ. D.,
WeisJ. J.,
TedderT. F.,
StromingerJ. L.,
BiroP. A.,
FearonD. T.1984; EpStein-Barr virus receptor of human B lymphocytes is the C3d receptor CR2. Proceedings of the National Academy of Sciences, U. S. A 81:4510–4514
FrakerP. J.,
SpeckJ. C.1978; Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1, 3, 4, 6-tetrachloro-3a, 6a-diphenylglycoluril. Biochemical and Biophysical Research Communications 80:849–857
GuttmanN.,
BaltimoreD.1977; A plasma membrane component able to bind and alter virions of poliovirus type 1: studies on cell-free alteration using a simplified assay. Virology 82:25–36
KitamuraN.,
SemlerB. L.,
RothbergP. G.,
LarsenG. R.,
AdlerC. J.,
DornerA. J.,
EminiE. A.,
HanecakR.,
LeeJ. J.,
Van der WerfS.,
AndersonC. W.,
WimmerE.1981; Primary structure, gene organization and polypeptide expression of poliovirus RNA. Nature, London 291:547–553
KochG.,
QuintrellN.,
BishopJ. M.1966; An agar cell-suspension plaque assay for isolated viral RNA. Biochemical and Biophysical Research Communications 24:304–309
KrahD. L.,
CrowellR. L.1982; A solid-phase assay of solubilized HeLa cell membrane receptors for binding group B coxsackieviruses and polioviruses. Virology 118:148–156
LebmanD.,
TruccoM.,
BotteroL.,
LangeB.,
PessanoS.,
RoveraG.1982; A monoclonal antibody that detects expression of transferrin receptor in human erythroid precursor cells. Blood 59:671–678
McdougalJ. S.,
BrowningS. W.,
KennedyS.,
MooreD. D.1983; Immunodot assay for determining the isotype and light chain type of murine monoclonal antibodies in unconcentrated hybridoma culture supernates. Journal of Immunological Methods 63:281–290
MinorP. D.,
SchildG. C.,
BootmanJ.,
EvansD. M. A.,
FergusonM.,
ReeveP.,
SpitzM.,
StanwayG.,
CannJ.,
HauptmannR.,
ClarkeL. D.,
MountfordR. C.,
AlmondJ. W.1983; Location and primary structure of a major antigenic site for poliovirus neutralization. Nature, London 301:674–679
MinorP. D.,
PipkinP. A.,
HockleyD.,
SchildG. C.,
AlmondJ. W.1984; Monoclonal antibodies which block cellular receptors of polioviruses. Virus Research 1:203–212
NodaM.,
SelingerZ.,
ScolnickE. M.,
BassinR. H.1983; Flat revertants isolated from Kirsten sarcoma virus-transformed cells are resistant to the action of specific oncogenes. Proceedings of the National Academy of Sciences, U. S. A 80:5602–5606
SchärliC. E.,
KochG.1984; Protein kinase activity in purified poliovirus particles and empty viral capsid preparations. Journal of General Virology 65:129–139
WychowskiC.,
Van der WerfS.,
SiffertO.,
CrainicR.,
BruneauP.,
GirardM.1983; A poliovirus type 1 neutralization epitope is located within amino acid residues 93 to 104 of viral capsid polypeptide VP1. EMBO Journal 2:2019–2024