Inhibition of rhinovirus attachment by neutralizing monoclonal antibodies and their Fab fragments.
Previous molecular and immunological studies have mapped four neutralization sites on human rhinovirus type 14 (B. Sherry, A. G. Mosser, R. J. Colonno, and R. R. Rueckert, J. Virol. 57:246-257, 1986). Eight monoclonal antibodies, one pair for each of the four target sites and all belonging to a single isotype, immunoglobulin G2a, were studied under conditions which resulted in 95% neutralization of infectious viral particles. All eight antibodies shifted the isoelectric point of virions from 6.7 to much more acidic forms, ranging from pI 1.8 to 3.2. In addition, antibodies targeted against three of the four neutralization sites caused significant aggregation of virions under the neutralization conditions employed. Aggregation could be reversed by digesting virus-antibody complexes with papain. Following papain digestion, the acidic pIs of three of the neutralized virus preparations returned to neutral and infectivity was restored. Membrane-binding assays with virus neutralized with a nonaggregating antibody showed a dose-related inhibition of virus attachment to cellular receptors. Purified Fab fragments at a 13- to 61-fold-higher concentration than intact antibodies caused a comparable isoelectric shift, neutralized virions in the absence of aggregation, and interfered with attachment of virions to host cell receptors in a membrane-binding assay. These findings suggest that neutralizing antibodies interfere with the attachment of rhinoviruses to cellular receptors and that bivalent attachment of antibody is not a prerequisite for neutralization.
Colonno
R J
RJ
Merck Sharp and Dohme Research Laboratories, West Point, Pennsylvania 19486.
Callahan
P L
PL
Leippe
D M
DM
Rueckert
R R
RR
Tomassini
J E
JE
D000911:Antibodies, Monoclonal / Q000276:immunology*; D001666:Binding Sites, Antibody; D002462:Cell Membrane / Q000382:microbiology; D004591:Electrophoresis, Polyacrylamide Gel; D006367:HeLa Cells; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology*; D007525:Isoelectric Focusing; D009500:Neutralization Tests; D010206:Papain / Q000494:pharmacology; D012229:Rhinovirus / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D014771:Virion / Q000187:drug effects / Q000378:metabolism
D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D010206:Papain