Open AccessStudy of neutralizing monoclonal antibodies to bovine herpes virus Type‐1 (Cooper strain) by immunoperoxidase and immunoelectron microscopy
Author(s) -
Winkler Maria Teresa C.,
Osorio Fernando A.,
Barahona Horácio J.,
Taffarel Marília
Publication year - 1995
Publication title -
fems immunology & medical microbiology
Language(s) - English
Resource type - Journals
eISSN - 1574-695X
pISSN - 0928-8244
DOI - 10.1111/j.1574-695x.1995.tb00071.x
Subject(s) - immunoelectron microscopy , immunoperoxidase , monoclonal antibody , biology , virology , microbiology and biotechnology , virus , cytoplasm , antibody , immunology
Abstract 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.