Study of neutralizing monoclonal antibodies to bovine herpes virus Type-1 (Cooper strain) by immunoperoxidase and immunoelectron microscopy |
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Authors: | Maria Teresa C Winkler Fernando A Osorio Horácio J Barahona Marília Taffarel |
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Institution: | CNPq MSc fellowship, Departamento de Microbiologia e Imunologia, Universidade Federal Rural do Rio de Janeiro, Itaguaí, RJ, Brazil;Centro Pan-Americano de Febre Aftosa, Duque de Caxias, RJ, Brazil;Departamento de Virologia, FIOCRUZ, Av. Brasil, 4365 - Manguinhos, C.P.: 926, CEP: 21045-900, Rio de Janeiro, RJ, Brazil |
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Abstract: | 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. |
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Keywords: | Bovine herpes virus Type-1 Infectious bovine rhinotracheitis Monoclonal antibody Immunoperoxidase Immunoelectron microscopy |
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