In vitro growth characteristics of bovine herpesvirus 4 (BHV-4) as revealed by indirect immunofluorescence assay with monoclonal antibodies and polyvalent antisera

Augsburger, H.; Metzler, A. E.

doi:10.1007/bf01315552

Cited by 12 publications

(3 citation statements)

References 22 publications

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“…The major glycoproteins identified with our MAbs exhibited similarities and differences in composition from those isolated from European strains [2,8]. The M r of the peptides 152, 123 and 46/46kDa of the glycoprotein complex that we described are similar to the glycoproteins GVP 6, 10, and 17 reported by Dubuisson et al [8].…”

Section: Discussionsupporting

confidence: 57%

Analysis of bovine herpesvirus 4 (DN 599) major antigens with monoclonal antibodies and polyclonal immune serum

Shen

Burger

et al. 1991

Archives of Virology

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Monoclonal antibodies (MAbs) and polyclonal immune sera were produced and used to identify the major antigens of bovine herpesvirus type 4 (BHV-4). SDS-polyacrylamide gel electrophoresis of immunoprecipitates of radiolabeled lysates from infected cells resolved 24 peptide bands varying from 12 kDa to over 300 kDa. Six peptides were identified as major viral antigens by immunoprecipitation. Based on the pattern of radioimmunoprecipitation, MAbs were assigned into four groups. Group 1 precipitated a tunicamycin-sensitive glycoprotein complex which contained six components (245, 190, 152, 123, and 48/46 kDa). Deglycosylation with endoglycosidase F revealed two peptides with Mr of 93 and 38 kDa as the basic peptides of the glycoprotein complex. In addition, a 115 kDa glycopeptide containing glycan-peptide bonds of mixed type was identified. Group 2 precipitated a non-glycosylated protein complex consisting of three monomers (33/31/30 kDa). Groups 3 and 4 reacted with single monomeric non-glycosylated peptides with Mr of 48 and 14 kDa, respectively. Although none of the MAbs exhibited significant neutralizing activity, some reacted strongly in immunosorbent and/or immunohistochemical assays, suggesting they may be good candidates for use in diagnostic assays.

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Section: Discussionsupporting

confidence: 57%

Analysis of bovine herpesvirus 4 (DN 599) major antigens with monoclonal antibodies and polyclonal immune serum

Shen

Burger

et al. 1991

Archives of Virology

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“…BHV-4-specific antibodies are demonstrated by complement fixation, dot immunobinding assay, agar gel immunodiffusion assay, immunofluorescence assay, or enzyme-linked immunosorbent assays (ELISAs) (10,13,32). The direct detection of the virus is based on virus isolation and characterization of specific letter-shaped inclusion bodies (1).…”

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confidence: 99%

Studies of in vivo distribution of bovine herpesvirus type 4 in the natural host

et al. 1996

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The in vivo distribution of bovine herpesvirus type 4 (BHV-4) was examined by testing nasal and conjunctival exudates, peripheral blood leukocytes, and various organs of experimentally infected calves. For virus detection, a nested PCR assay, virus isolation, and immunohistochemistry were applied. The nervous system and the muscles were free of viral DNA. Liver and intestinal lymph nodes contained low amounts of virus (less than two copies per 1 g of cellular DNA). Intestinal, tonsil, thymus, and kidney tissues contained more viral DNA copies (5 to 50 copies per 1 g of cellular DNA). The highest amounts of BHV-4 DNA (50 to 500 copies per 1 g of cellular DNA) were found in the spleen, lungs, trachea, and nasal epithelium. Amplification of DNA from blood lymphocytes through postinoculation (p.i.) day 48 proved that the virus started to replicate in these cells immediately after inoculation of the calves and that intensive virus growth took place during the 7 to 8 weeks of the infection. The number of virus-infected lymphocytes reached the maximum on p.i. days 22 to 26 and slowly declined thereafter. Virus-infected cells were found only in the spleen on p.i. day 48 by immunohistochemistry. Western blotting (immunoblotting) detected signs of an immune response against 9 of the 29 BHV-4 proteins.

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“…Consequently, the effects of pBo17 activity expression in vivo should vary based on the cell type infected; unless the infection occurs in cells which do not express b1,6GnT activities or unless BoHV-4 infection completely down regulates the expression of host cell b1,6GnT activities. The latter hypothesis is unlikely because BoHV-4 does not affect host cell protein expression (Augsburger & Metzler, 1989). However, several hypotheses that are not mutually exclusive could be made concerning the role of Bo17 in the biology of BoHV-4 infection.…”

Section: Discussionmentioning

confidence: 99%

The core 2 β-1,6-N-acetylglucosaminyltransferase-M encoded by bovine herpesvirus 4 is not essential for virus replication despite contributing to post-translational modifications of structural proteins

Markine-Goriaynoff

Gillet

Karlsen

et al. 2004

Journal of General Virology

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The Bo17 gene of bovine herpesvirus 4 (BoHV-4) is the only virus gene known to date that encodes a homologue of the cellular core 2 b-1,6-N-acetylglucosaminyltransferase-mucine type (C2GnT-M). Recently, our phylogenetic study revealed that the Bo17 gene has been acquired from an ancestor of the African buffalo around 1?5 million years ago. Despite this recent origin, the Bo17 sequence has spread to fixation in the virus population possibly by natural selection. Supporting the latter hypothesis, it has been shown by our group for the V. test strain that Bo17 is expressed during BoHV-4 replication in vitro, and that Bo17 expression product (pBo17) has all three enzymic activities exhibited by cellular C2GnT-M, i.e. core 2, core 4 and I branching activities. In the present study, firstly it was investigated whether encoding a functional C2GnT-M is a general property of BoHV-4 strains. Analysis of nine representative strains of the BoHV-4 species revealed that all of them express the Bo17 gene and the associated core 2 branching activity during virus replication in vitro. Secondly, in order to investigate the roles of Bo17, its kinetic class of expression was analysed and a deleted recombinant strain was produced. These experiments revealed that Bo17 is expressed as an early gene which is not essential for virus replication in vitro. However, comparison of the structural proteins, produced by the wild-type, the revertant and the deleted viruses, by 2D gels demonstrated that pBo17 contributes to the post-translational modifications of structural proteins. Possible roles of Bo17 in vivo are discussed.

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In vitro growth characteristics of bovine herpesvirus 4 (BHV-4) as revealed by indirect immunofluorescence assay with monoclonal antibodies and polyvalent antisera

Cited by 12 publications

References 22 publications

Analysis of bovine herpesvirus 4 (DN 599) major antigens with monoclonal antibodies and polyclonal immune serum

Analysis of bovine herpesvirus 4 (DN 599) major antigens with monoclonal antibodies and polyclonal immune serum

Studies of in vivo distribution of bovine herpesvirus type 4 in the natural host

The core 2 β-1,6-N-acetylglucosaminyltransferase-M encoded by bovine herpesvirus 4 is not essential for virus replication despite contributing to post-translational modifications of structural proteins

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