Effects of specific monoclonal antibodies on La Crosse virus neutralization: aggregation, inactivation by Fab fragments, and inhibition of attachment to baby hamster kidney cells.
At high concentrations, several monoclonal antibodies to the G1 glycoprotein of La Crosse (LAC) virus aggregated the virus. To determine whether this accounted for the neutralization, the monoclonal antibodies were digested to make Fab fragments. With one exception, each monovalent antibody neutralized LAC virus to the same extent that bivalent antibody did, although higher concentrations were needed. Fab fragments of synergistic pairs of antibodies also exhibited enhanced binding in a competition binding assay but did not increase neutralization. To determine specific mechanisms for neutralization, the effects of polyclonal or monoclonal antibodies on virus attachment were examined. Polyclonal antibody to LAC virus reduced virus attachment by only 68% although it neutralized 99.99% of the virus. When virus was preincubated with a neutralizing monoclonal antibody to each of seven antigenic regions on G1, only antibody to one region reduced attachment of virus by as much as 92%. Antibodies to two regions that neutralize virus by 90-98% only inhibited attachment by 9 and 13%, respectively. The other antibodies showed intermediate degrees of neutralization and inhibition of attachment. Pairs of antibodies previously shown to be synergistic in neutralizing activity did not inhibit attachment any more than the single antibodies did.
Department of Microbiology, California State University, Long Beach 90840.
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Kingsford
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Boucquey
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Cardoso
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D001667:Binding, Competitive; D002460:Cell Line; D006224:Cricetinae; D002141:Encephalitis Virus, California / Q000276:immunology* / Q000502:physiology; D007140:Immunoglobulin Fab Fragments / Q000276:immunology*; D007668:Kidney; D007700:Kinetics; D009500:Neutralization Tests; D005719:gamma-Globulins / Q000276:immunology
D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D005719:gamma-Globulins