Analysis of bovine herpesvirus 4 (DN 599) major antigens with monoclonal antibodies and polyclonal immune serum

Abstract

Monoclonal antibodies (MAbs) and polyclonal immune sera were produced and used to identify the major antigens of bovine herpesvirus type 4 (BHV-4). SDS-polyacrylamide gel electrophoresis of immunoprecipitates of radiolabeled lysates from infected cells resolved 24 peptide bands varying from 12 kDa to over 300 kDa. Six peptides were identified as major viral antigens by immunoprecipitation. Based on the pattern of radioimmunoprecipitation, MAbs were assigned into four groups. Group 1 precipitated a tunicamycin-sensitive glycoprotein complex which contained six components (245, 190, 152, 123, and 48/46 kDa). Deglycosylation with endoglycosidase F revealed two peptides with Mr of 93 and 38 kDa as the basic peptides of the glycoprotein complex. In addition, a 115 kDa glycopeptide containing glycan-peptide bonds of mixed type was identified. Group 2 precipitated a non-glycosylated protein complex consisting of three monomers (33/31/30 kDa). Groups 3 and 4 reacted with single monomeric non-glycosylated peptides with Mr of 48 and 14 kDa, respectively. Although none of the MAbs exhibited significant neutralizing activity, some reacted strongly in immunosorbent and/or immunohistochemical assays, suggesting they may be good candidates for use in diagnostic assays.