Emilie Weiland

LOCALIZATION OF PESTIVIRAL ENVELOPE PROTEINS ERNS AND E2 AT THE CELL SURFACE AND ON ISOLATED PARTICLES

The glycoproteins E(rns) of classical swine fever virus (CSFV) and E(rns) and E2 of bovine viral diarrhoea virus (BVDV) are shown to be located at the surface of infected cells by the use of indirect immunofluorescence and by cytofluorometric analysis. The positive immunostaining of the cell surface was further analysed by immunogold electron microscopy and it could be shown that only extracellular virions were labelled. Gold granules were not seen at the cellular plasma membrane. In contrast to BVDV E2, the CSFV E2 of virions sticking to the plasma membrane was not accessible to the respective monoclonal antibodies. However, CSFV particles isolated from culture supernatant were able to bind both monoclonal anti-E(rns) and anti-E2 antibodies. For CSFV and BVDV, binding of anti-E(rns) antibodies to the virions was more pronounced than that of anti-E2. This finding was unexpected since E2 is considered to be the immunodominant glycoprotein.

Porcine reproductive and respiratory syndrome virus (PRRSV): monoclonal antibodies detect common epitopes on two viral proteins of European and U.S. isolates.

Sixteen hybridoma cell lines secreting monoclonal antibodies (mAbs) directed against two dutch isolates of the causative virus of Porcine Reproductive and Respiratory Syndrome (PRRSV) were produced. The hybridoma cells resulted from fusions of SP2/0 myeloma cells with splenocytes of STU mice immunized with purified PRRSV after induction of immunotolerance against host cell constituents. Screening of supernatant fluids was performed by an indirect immunofluorescence assay on PRRSV-infected porcine alveolar macrophages. Immunoblotting studies revealed that the mAbs had different protein specificities. One mAb reacted with a viral 15 kD protein, eleven were directed against a 40-50 kD protein, and four against a 30-40 kD protein. The mAb against the 15 kD putative nucleocapsid protein as well as five mAbs against the 40-50 kD protein recognized epitopes on these proteins which are conserved in various European and U.S. isolates of PRRSV.

Lactate dehydrogenase-elevating virus induces antibodies reactive with a surface antigen of aetiologically unrelated murine cell transformants

Mice infected with lactate dehydrogenase-elevating virus (LDV) developed antibodies reactive with a tumour cell surface antigen (TSA) of Moloney sarcoma virus (Mo-MSV)-transformed mouse cells (Sac). We demonstrate that STU mice infected with LDV were protected against growth of syngeneic Sac tumour cells as early as 23 days post-infection (p.i.) and up to 5 months p.i. Nine LDV strains, including the neurovirulent LDV-C, elicited production of anti-TSA antibodies, which were restricted to the IgM isotype. Monoclonal anti-TSA antibodies were raised 4 days after infection of STU mice with LDV. When tested against several transformants of STU and BALB/c mouse origin they were found to react with Mo-MSV transformants (PV-TC-77, STU mouse origin; MSV85 C1 3, BALB/c mouse origin), methylcholanthrene-transformed MethA cells (BALB/c origin) and L929 cells. We suggest that the well known tumour growth inhibition by LDV is due to LDV-induced anti-TSA antibodies.