Glycoprotein E of bovine herpesvirus type 1 is involved in virus transmission by direct cell-to-cell spread.

In order to identify the role of the bovine herpesvirus type 1 (BHV-1) glycoprotein E (gE) in the viral infection cycle, we have constructed a BHV-1 gE deletion mutant strain (BHV-1 gE-). This strain was assayed in vitro by comparing its growth kinetics with the wild type strain used as a host of the deletion. Our results indicate that those conditions which prevent the infection by direct adsorption to the cells (presence of a semi-solid medium or presence of neutralizing antibodies in the medium) selectively inhibit the growth of the gE- strain, suggesting that gE plays a central role in the BHV-1 spread by direct cell-to-cell transmission, a major mechanism of the BHV-1 in vivo virulence.

Resistance and susceptibility of bovine cells expressing herpesviral glycoprotein D homologs to herpesviral infections.

Bovine cell lines individually expressing two related herpesviral proteins, pseudorabies virus glycoprotein 50 and herpes simplex virus type 1 glycoprotein D, were examined for their susceptibility/resistance to infection with several alphaherpesviruses. Cell lines expressing gp50 or gD-1 resisted plaque formation by the homologous virus more than by the heterologous viruses. Bovine cells expressing bovine herpesvirus 1 glycoprotein IV (gIV) were susceptible to infection with three other bovine herpesviruses: bovine herpesvirus 2, bovine herpesvirus 4 (BHV-4) and alcelaphine herpesvirus 1. One line of gIV-expressing cells was resistant to the formation of BHV-4 plaques, suggesting that a cell-associated factor may be responsible for inhibiting cell-to-cell spread.

A comparison of the acid-soluble polypeptides of five herpesviruses.

The polypeptides soluble in 0.25 M-HCl were extracted from the nuclei of BHK cells infected with herpes simplex virus type 1 or type 2 and separated by SDS-PAGE. Seventeen polypeptides were detectable in each extract of which 10 type 1 and nine type 2 polypeptides were reproducibly effectively extracted. In cells infected with bovine mammillitis virus, pseudorabies virus or equine herpesvirus type 1, at least 12, 13 and eight polypeptides respectively were acid-soluble. In addition to histones, three other cellular polypeptides were present in sizeable quantities in the acid extracts and could obscure other acid-soluble viral polypeptides. Possible relationships between some polypeptides of the five herpesviruses are discussed.

Local tissue temperature: a critical factor in the pathogenesis of bovid herpesvirus 2.

The effect of local tissue temperature on bovid herpesvirus 2 lesions in bovine skin was studied. Two areas of thoracic skin were multiply inoculated with virus and maintained at different temperatures; one above and the other below the rectal temperature of the animal. An entire inoculation site was removed daily from each area and subjected to virus, viral antigen, and interferon titrations and both light and electron microscopic examinations. Lesions in cold skin appeared sooner after inoculation, were larger and deeper, contained more infectious virus, viral antigen, and interferon, and lasted longer than lesions in hot skin. Differences in viral titers greater than 10(9) 50% tissue culture infective doses per gram were measured on postinfection days 5 to 9. These studies have demonstrated for the first time that temperature effects on viral pathogenesis may operate at the local level rather than by systemic modification of immune responses. The results also suggest that despite the presence of virus in internal organs of bovid herpesvirus 2-infected cattle and the ability of bovid herpesvirus 2 to replicate to very high titers in these tissues in vitro, only the skin is cool enough to permit substantial viral replication and dermal lesions in vivo. The observed restriction of bovid herpesvirus 2 skin lesions to the udder and teats of cattle and the restriction of outbreaks of disease to months when the temperature is declining may also be reflections of this temperature sensitivity.

Reactivation of Bovid herpesvirus 1 and 2 and parainfluenza-3 virus in calves latently infected.

Two experiments were carried out to determine whether Bovid herpesvirus (BHV) 2 is able to induce a recurrent infection in experimentally infected calves. In the first experiment the stress induced by dexamethasone (DMS) treatment failed to reactivate the clinical condition or to induce shedding of BHV2. However, treatment with DMS reactivated a latent BHV1 infection in all calves previously inoculated with BHV2 and also in two noninoculated controls. Probably, because of the interference by BHV1 the study failed to resolve the question as to whether BHV2 could induce a recurrent infection. Consequently, a second experiment was performed using calves devoid of antibody to BHV1 and, therefore, probably, free of virus. By this study it was demonstrated that BHV2 can remain as a latent infection in cattle, which, when immunosuppressed as with DMS, can be reactivated. A finding of considerable interest in this experiment was that in 1 calf a concurrent piroplasma infection was also, unexpectedly, discovered. Recrudescence of latent BHV1 infection was induced by DMS treatment of calves possessing antibody to the virus. The infection once reactivated, was readily transmitted by contact to three other calves devoid of antibody to BHV1. In the same experiment Parainfluenza-3 (PI-3) virus was unexpectedly isolated from all calves. It was speculated that all calves were latently infected with PI-3 virus with concurrent infection by HBV1 acting as a stress inducing PI-3 reactivation. These studies seem to indicate that mixed infections could have an important role in the mechanism involved in the establishment of latent infections and viral reactivation.

Bovid herpesvirus 2: natural spread among breeding bulls.

Sera collected over a seven year period, together with detailed health and management records, were used in a retrospective study of Bovid herpesvirus 2 (BHV-2) spread within a large herd of breeding bulls. Virus spread rapidly throughout the bulls in one barn. Transfer of infected bulls to other barns was followed by further, although sporadic, spread of virus. Spread of BHV-2 from seropositive animals to susceptible bulls in close contact led to the conclusion that reactivation and transmission of latent virus had occurred. Semen from seropositive bulls did not transmit BHV-2 to seronegative calves that were purposefully inoculated by the intranasal or intravenous routes.

Latent infection of cattle with bovid herpesvirus 2.

In a series of experiments, 10 calves were infected by different routes with bovine mammillitis strain of bovid herpesvirus 2 (BHV2). Nine of the calves developed lesions and BHV2 was isolated from 8 calves. At intervals, after clinical recovery, the calves were given a course of corticosteroid by intravenous injection. Subsequently 7 calves developed lesions and from 3 calves BHV2 was reisolated. The shortest interval between initial infection and reisolation of virus was 10 weeks. In one calf virus was recovered on 3 occasions, at intervals of 20, 44 and 74 weeks post-infection. It is probable that BHV2 can occur as a latent infection in calves and that BHV2 should be included among the herpesviruses known to remain latent.

A comparison in calves of the antigenicity of three strains of bovid herpesvirus 2.

Three strains of bovid herpesvirus 2, viz. Allerton, bovine mammillitis and 69/1LO were used to infect calves intradermally. Twenty-eight days later the immunity of the calves was challenged by intravenous injection of a homologous or heterologous strain. Challenge control calves developed a fever (greater than 40 degrees C) lasting several days and widespread skin lesions which varied with the strain. Homologous challenge of the primary infection produced neither skin lesions nor febrile response, except in one calf in which fever was noted on one day. Heterologous challenge did not cause skin lesions but fever occurred in 8/12 calves. In particular Allerton virus failed to protect completely against heterologous challenge. Despite minor differences evident in these experiments, it is recommended that these isolates should be considered as strains of the same virus--bovid herpesvirus 2.

Experimental studies concerning the possibility of a latent carrier state in bovine herpes mammillitis (BHM).

Adult bovines, young calves and young rats were infected with bovine herpes mammillitis (BHM) virus providing a group of BHM-recovered animals in which to investigate a possible latent carrier state. The progression of the acute infection and serological response was essentially similar to that previously reported. The effect of natural (calving, littering) and artificial (corticosteroid treatment) stress on these animals following recovery from infection was investigated, but reactivation of BHM was never detected. Long term observation of cocultures of BHM virus infected-and-recovered bovine teat dermis with bovine embryo cells failed to reveal any evidence for the induction of BHM virus replication even in those cocultures treated with mitomycin C or bromodeoxyuridine. Similarly, cocultures utilizing lumbar dorsal root ganglia from the same animals were also negative with respect to the induction of virus replication. Experiments using cytosine arabinoside indicated that an artificial in vitro latent carrier state of BHM virus could be maintained for 6 days with subsequent sporadic virus reactivation, but an attempt to maintain such a state for 12 days was unsuccessful, with no subsequent virus reactivation.