Abstract
The transmission of Macacine alphaherpesvirus 1 (McHV-1) from macaques, the natural host, to humans causes encephalitis. In contrast, human infection with Cercopithecine alphaherpesvirus 2 (CeHV-2), a closely related alphaherpesvirus from African vervet monkeys and baboons, has not been reported and it is believed that CeHV-2 is apathogenic in humans. The reasons for the differential neurovirulence of McHV-1 and CeHV-2 have not been explored on a molecular level, in part due to the absence of systems for the production of recombinant viruses. Here, we report the generation of a fosmid-based system for rescue of recombinant CeHV-2. Moreover, we show that, in this system, recombineering can be used to equip CeHV-2 with reporter genes. The recombinant CeHV-2 viruses replicated with the same efficiency as uncloned, wt virus and allowed the identification of cell lines that are highly susceptible to CeHV-2 infection. Collectively, we report a system that allows rescue and genetic modification of CeHV-2 and likely other alphaherpesviruses. This system should aid future analysis of CeHV-2 biology.
Highlights
Herpesviruses are a large family of DNA viruses that usually establish a lifelong latency or persistence in their hosts
Viral DNA was isolated from isolated from virions, sheared to fragments of 30–40 kb, and size fractionated on a low percentage virions, sheared to fragments of 30–40 kb, and size fractionated on a low percentage agarose gel agarose gel (Figure 1a)
We describe a fosmid-based system for the generation of recombinant Cercopithecine alphaherpesvirus 2 (CeHV-2). This system allowed the rescue of recombinant CeHV-2 that replicated with the same efficiency as uncloned wt virus
Summary
Herpesviruses are a large family of DNA viruses that usually establish a lifelong latency or persistence in their hosts. Herpes simplex virus 1 (HSV-1, human alphaherpesvirus 1) and other members of the genus Simplexvirus, subfamily Alphaherpesvirinae, establish latency in sensory neurons and have a conserved genome structure. Primate simplexviruses coevolved with their host species, resulting in codivergent trees for the viruses and their respective host species [1,2]. Despite this coevolution, these viruses are not species-specific, and cross-species transmission has been frequently observed. The transmission of Macacine alphaherpesvirus 1 (McHV-1, termed herpes B virus). Viruses 2019, 11, 1026 from macaques to humans causes encephalitis that is associated with a high case fatality rate in the absence of treatment [3]. HSV-1 rarely causes encephalitis and the McHV-1-related viruses
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