A Neutralizing Monoclonal Antibody to Bovine Rotavirus VP8 Neutralizes Rotavirus Infection Without Inhibiting Virus Attachment to MA-104 Cells

Joongbok Lee, Lorne A. Babiuk, Dongwan Yoo

Research output: Contribution to journalArticlepeer-review

Abstract

VP8*, the N-terminal cleavage product of rotavirus VP4, contains the virus neutralizing epitopes in the hemagglutination domain. To map the neutralizing epitope, we developed monoclonal antibodies specific for VP4 of bovine rotavirus C486 (BRV). A neutralizing escape mutant was generated by one of these monoclonal antibodies (2E8) and a point mutation (Glu → Asp) was found at aa 116 of VP8*. To investigate the effect of this mutation on the cellular binding and hemagglutination activities, the VP8* genes of the escape mutant and wild type (WT) virus were expressed in E. coli and their functional activities were compared. Both the escape mutant and WT virus VP8* showed hemagglutination and MA-104 cell binding activities. However, hemagglutination activity of the WT virus VP8* was inhibited by 2E8, but that of the escape mutant VP8* was not. These data indicate that the neutralizing epitope is located in the HA domain but is not critical for rotavirus attachment to MA-104 cells. To understand virus neutralization, radiolabelled BRV was incubated with 2E8 and the distribution of radioactivity in a CsCl density gradient was analysed as was the morphology of the virions in peak fractions. Interaction of 2E8 with rotavirus led to virus morphological changes with a concomitant shift in buoyant density. These data suggest that aa 116 influences the binding of 2E8 which in turn may alter virus integrity.

Original languageEnglish (US)
Pages (from-to)63-67
Number of pages5
JournalCanadian Journal of Veterinary Research
Volume62
Issue number1
StatePublished - 1998
Externally publishedYes

ASJC Scopus subject areas

  • veterinary(all)

Fingerprint Dive into the research topics of 'A Neutralizing Monoclonal Antibody to Bovine Rotavirus VP8 Neutralizes Rotavirus Infection Without Inhibiting Virus Attachment to MA-104 Cells'. Together they form a unique fingerprint.

Cite this