Comparison of Ruminant Alphaherpesviruses Serologically Related to Bovine Herpesvirus-1

Lyaku, Japhet Robert (1993) Comparison of Ruminant Alphaherpesviruses Serologically Related to Bovine Herpesvirus-1. PhD thesis, University of Glasgow.

Abstract

Isolates of bovine herpesvirus-1 subtype 1 (BHV-1.1) and bovine herpesvirus-1 subtype 2 (BHV-1.2), the cause of infectious bovine rhinotracheitis-infectious pustular vulvovaginitis were compared with three other distinct ruminant alphaherpesviruses isolated from a goat (CapHV-2), a red deer (CerHV-1) and a reindeer (RanHV-1). Restriction endonuclease profiles produced by BglII differentiated the five ruminant alphaherpesviruses. Amplification of a 468 by fragment of each of the five alphaherpesviruses by the polymerase chain reaction (PCR) was achieved using 22 by primers selected from the BHV-1 glycoprotein gI DNA sequence. All the amplified products contained BglI and HinfI cleavage sites. Only RanHV-1 did not contain Smal or Aval sites. Serological comparison by ELISA and serum neutralisation (SN) tests using rabbit hyperimmune sera, and cattle, goat, red deer and reindeer convalescent sera revealed a close serological cross-reactivity between all 5 viruses. Analysis of their relationships confirmed that: 1) the two BHV-1 viruses were most closely related, 2) CerHV-1 and CapHV-2 were more closely related to BHV-1 than they were to each other, and 3) RanHV-1 was more related to CerHV-1 than to BHV-1 and CapHV-2. Western blotting using polyclonal sera identified at least seven major proteins common to all the viruses with molecular weights of 130kDa, 108 kDa, 90 kDa, 87 kDa, 74 kDa, 64 kDa and 45 kDa. With a view to identifying epitopes which would allow the differentiation of these viruses, a panel of 14 monoclonal antibodies (Mabs) against CerHV-1 were derived from BALB/c mice previously immunised with gradient purified CerHV-l. Four Mabs which had neutralising activity against CerHV-1 reacted in Western blots with polypeptides of Mi. 68-70 kDa and 74 kDa. The other 10 Mabs reacted in Western blots with polypeptides of Mi. 68-70 kDa and/or 64 kDa. Radioimmunoprecipitation (RIP) results indicated that the 74 kDa protein detected in CerHV-1 was analogous to the 74 kDa cleavage product of the gI glycoprotein of BHV-1. The reactivity of each Mab against the five ruminant alphaherpesviruses was tested by ELISA. Two Mabs cross-reacted with all the five ruminant alphaherpesviruses while 10 others showed variable reactivity with the 5 viruses. Two Mabs reacted exclusively with CerHV-1 antigens and these may be suitable for developing a diagnostic test to distinguish antibodies between BHV-1 and CerHV-1 in their natural hosts.

• Item Type: Thesis (PhD)
• Qualification Level: Doctoral
• Additional Information: Adviser: P F Nettleton
• Keywords: Virology, Animal diseases
• Date of Award: 1993
• Depositing User: Enlighten Team
• Unique ID: glathesis:1993-76380
• Copyright: Copyright of this thesis is held by the author.
• Date Deposited: 19 Nov 2019 14:47
• Last Modified: 19 Nov 2019 14:47
• URI: https://theses.gla.ac.uk/id/eprint/76380