Abstract
Antigens of a bovine herpesvirus type 5 (BHV-5), isolated from a cow with a neurological infection in Rio Grande do Sul State, Brazil, were used to immunize BALB/c mice to produce monoclonal antibodies (mAbs). Eleven hybridomas secreting mAbs directed at BHV-5 antigens were obtained after two fusions and screening of 356 hypoxanthine-aminopterin-thymidine-resistant clones. The mAbs reacted at dilutions up to 1:500 (hybridoma culture supernatant) and up to >1:10,000 (ascitic fluid) in an indirect fluorescent antibody assay (IFA) and in immunoperoxidase staining of BHV-5-infected cells. Four mAbs (1D12, 2E2, 2G10 and 4E4) showed virus-neutralizing activity against the parental BHV-5 isolate. Five mAbs (1F3, 2A6, 2F9, 2G10 and HB24L) reacted in Western immunoblotting with a protein of approximately 90 kDa. Three other mAbs (2E2, 3D6 and 4E4) reacted in IFA with antigens of a BHV-1 mutant glycoprotein C- negative strain, demonstrating that they are directed at a viral antigen other than glycoprotein C. The eleven mAbs tested reacted with 20 BHV-5 field isolates and nine mAbs reacted with 10 BHV-1 isolates. Two mAbs (1F3 and 2F9) failed to react with BHV-1 field isolates, although they displayed a weak and nonreproducible reaction with the BHV-1 reference strain Los Angeles. These mAbs may be very useful in distinguishing between BHV-1 and BHV-5 infections since most of the traditional reagents and techniques are unable to do so. One mAb (2F9) was shown to bind to viral antigens by immunohistochemistry of histological sections of the brain of a BHV-5-infected calf. These results demonstrate that the mAbs produced here are suitable for use in a variety of immunological techniques and therefore may be useful for diagnostic and research purposes.
Highlights
Bovine herpesvirus type 5 (BHV-5) is an α-herpesvirus associated with fatal meningoencephalitis in cattle [1]
These monoclonal antibodies (mAbs) were tested for the ability to bind to viral antigens in MDBK cells infected with a glycoprotein C-deleted bovine herpesvirus type 1 (BHV1) mutant (IBRVdlTKdlgIII) [23] by indirect fluorescent antibody assay (IFA)
We report the production of 11 mAbs against antigens of a Brazilian BHV-5 isolate
Summary
Bovine herpesvirus type 5 (BHV-5) is an α-herpesvirus associated with fatal meningoencephalitis in cattle [1]. In spite of the differences in the envelope glycoproteins, the traditional serological techniques and the majority of BHV-1 and BHV-5 monoclonal antibodies (mAbs) are unable to distinguish between these two viruses [16,17]. The prevalence and geographical distribution of BHV-5 infection in cattle populations are largely unknown This is due, in part, to the widespread distribution of BHV-1 infection and vaccination, associated with the inability of traditional techniques to distinguish between BHV-1- and BHV-5-induced antibodies. In this respect, the availability of reagents and techniques to identify and differentiate between BHV-1 and BHV-5 would be of great value for diagnostic and epidemiological purposes. Most of these mAbs recognized a panel of BHV-1 and BHV5 field isolates and two mAbs were able to differentiate between BHV-1 and BHV-5
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