Study of neutralizing monoclonal antibodies to bovine herpes virus type-1 (Cooper strain) by immunoperoxidase and immunoelectron microscopy.
Three neutralizing monoclonal antibodies (mAbs) that are specific against bovine herpes virus Type-1 (BHV-1) were studied as to their viral specificity by immunoperoxidase and immunoelectron microscopy. Microscopic examination of GBK BHV-1 infected cells revealed peroxidase activity represented by red-brown granular deposits in the nucleus and cytoplasm. No immunoperoxidase activity was observed in negative controls. For the ultrastructural observations, two approaches were used. Firstly we tested a pre-embedding technique using GBK infected cells, mAbs and gold conjugated-protein A. Gold particles were observed linked to the viral envelopes and to the host cell membrane. Alternatively, a second technique employed BHV-1 purified by potassium tartrate gradients, mAbs and gold conjugated-protein A. After performing the immune reaction, the samples were adsorbed to formvar-coated grids, stained with phosphotungstic acid and observed in a transmission electron microscope. Gold particles were mainly attached to the virion envelope.
Winkler
M T
MT
Departamento de Microbiologia e Imunologia, Universidade Federal Rural do Rio de Janeiro, Itaguaí, Brazil.
Osorio
F A
FA
Barahona
H J
HJ
Taffarel
M
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis*; D000914:Antibodies, Viral / Q000032:analysis*; D000918:Antibody Specificity; D001426:Bacterial Proteins; D002213:Capsid / Q000276:immunology / Q000648:ultrastructure; D002417:Cattle; D002462:Cell Membrane / Q000276:immunology / Q000648:ultrastructure; D002467:Cell Nucleus / Q000276:immunology / Q000648:ultrastructure; D002478:Cells, Cultured; D017979:Gold Colloid; D006566:Herpesviridae Infections / Q000276:immunology* / Q000821:virology; D007242:Herpesvirus 1, Bovine / Q000145:classification / Q000276:immunology* / Q000302:isolation & purification; D007124:Immunoenzyme Techniques; D007668:Kidney / Q000276:immunology / Q000648:ultrastructure; D016253:Microscopy, Immunoelectron; D009500:Neutralization Tests
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D001426:Bacterial Proteins; D017979:Gold Colloid; C070001:protein AG-gold complex