Antigens of infectious laryngotracheitis herpesvirus defined by monoclonal antibodies

Abstract

Monoclonal antibodies to glycoprotein and protein antigens of infectious laryngotracheitis virus (ILTV) were divided into five groups on the basis of their reactivity in immunofluorescence and Western blotting. Group I antibodies recognised a single band of 60 K and Group II antibodies recognised bands of 205, 160, 115, 90 and 85 k in Western blotting. In immunofluorescence both these groups of antibodies reacted with antigens located in the cytoplasm of fixed virus-infected cells and they also reacted with unfixed cells, suggesting that these antigens are on the surface of virus-infected cells. While Group I monoclonal antibodies did not react with extracts of tunicamycin-treated cells, some Group II antibodies recognised bands of decreased molecular weight compared to those present in untreated cells. The reactivity of the Group II antibodies with extracts of tunicamycin-treated cells suggested that they recognised at least three different epitopes which was confirmed by ELISA additivity assays. Monoclonal antibodies of Group III, Group IV and Group V recognised several low molecular weight proteins from 45 to 24 k. Immunofluorescence studies showed that these were nuclear and cytoplasmic antigens that were not present on the surface of virus-infected cells.