Expression of Epstein-Barr virus nuclear antigen 1,2A and 2B in the baculovirus expression system: serological evaluation of human antibodies to these proteins

Abstract

The Epstein-Barr virus encoded nuclear antigens 1,2A, and 2B (EBNA 1, EBNA 2A, and EBNA 2B) were expressed in a baculovirus system. The full length recombinant proteins were recognized by polyclonal rabbit sera and by human sera. An immunofluorescence (IF) test for the differentiation between EBNA 1 and EBNA 2 antibodies in human sera was established with the expressed proteins. None of 55 sera of patients with infectious mononucleosis (IM) had anti-EBNA 1 antibodies while 36 of these sera had anti-EBNA 2A antibodies. Conversely, 47 of 51 sera from EBV-positive healthy carriers had anti-EBNA 1 antibodies and 18 of these sera had anti-EBNA 2A antibodies. The sensitivity and specificity of the EBNA 1 IF for the diagnosis of IM were higher as compared to conventional anti-complement immunofluorescence (ACIF). In the IF test differentiation between type A and type B EBV infection was only possible in sera from the IM patients. An immunoblot (IB) with low amounts of baculovirus expressed EBNA 2A and EBNA 2B antigen was carried out. Twenty-nine of 31 sera from IM patients or from healthy carriers with EBNA 2 antibodies reacted predominantly with EBNA 2A, whereas a known type B serum reacted strongly with EBNA 2B than with EBNA 2A.