The primary production of an infectious recombinant Herpes Simplex Virus vaccine

O'Keeffe, R. S.; Johnston, M. D.; Slater, Nigel K.H.

doi:10.1002/(sici)1097-0290(19980205)57:3<262::aid-bit2>3.0.co;2-f

Cited by 19 publications

(20 citation statements)

References 21 publications

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“…No freeze-thaw step was performed. 22 The supernatant was carefully removed and passed through a 0.8-m fine porosity filter (Nalgene, Rochester, NY) to exclude residual cells and debris. Virus was centrifuged at 17 700g in a JA-10 rotor (Beckman, Palo Alto, CA) for 2 hours.…”

Section: Virus Preparation and Labelingmentioning

confidence: 70%

Characterization of Kaposi sarcoma–associated herpesvirus/human herpesvirus–8 infection of human vascular endothelial cells: early events

Dezube

Zambela

Sage

et al. 2002

Blood

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IntroductionKaposi sarcoma-associated herpesvirus (KSHV)/human herpesvirus-8 (HHV-8) is the first human ␥-herpesvirus identified since the 1964 discovery of Epstein-Barr virus (EBV). 1,2 HHV-8 has been causally implicated in the pathogenesis of 3 proliferative disorders that occur with increased frequency in the immunocompromised host: Kaposi sarcoma (KS), primary effusion lymphoma (PEL), and the plasmablastic variant of multicentric Castleman disease (MCD). [1][2][3][4] Although KS is uncommon, it is the most common malignancy associated with human immunodeficiency virus infection. KS lesions may spontaneously resolve following immune reconstitution or may wax and wane with an individual's state of T-cell competence. 5,6 This remarkable feature, shared by EBVassociated B-cell lymphoproliferative disease, suggests that a latent reservoir of HHV-8 infection in vivo gives rise to KS.Many HHV-8-encoded proteins, which contribute to the pathogenesis of KS, PEL, and MCD, are homologs of cellular genes involved in inflammation, cell cycle regulation, and angiogenesis. 7 Two such proteins are those encoded by open-reading frame (ORF)-74 and ORF72. The respective proteins are expressed in distinct phases of the virus life cycle. 7 ORF74 encodes a lytic viral G protein-coupled receptor (vGPCR), which is homologous to human interleukin-8 receptors. 7,8 When injected into mice, vGPCRtransfected rodent fibroblasts cause spindle cell tumors with prominent vasculature. 7,9 ORF72 encodes a latent viral cyclin that can activate kinases that inactivate retinoblastoma protein (pRB), a checkpoint protein that inhibits entry into S phase. 7,10 In the complex lesions of KS, viral gene products are expressed in endothelial cells, in spindle cells of probable endothelial origin, in monocytes, and possibly in some infiltrating lymphocytes. The endothelial cells primarily harbor virus as circular episomes and synthesize latency-associated gene products, whereas the smaller monocyte subpopulation is productively infected. 11,12 Normal primary endothelial cells can be infected with HHV-8 in vitro. However, the efficiency and outcome of infections have been variable despite diverse efforts to optimize in vitro culture. [13][14][15][16][17] In contrast, herpesvirus infection of relevant primary cells, whether productive or causing immortalization, is usually robust. Thus, the recent studies of endothelial cell infection raise several questions concerning whether a more physiologic target cell remains to be identified, whether variable transmission in vitro is caused by technical difficulties, whether in vitro infection is blocked at a particular phase of the virus life cycle, or whether indeed HHV-8 infection is inefficient when compared with infection by other members of the human herpesvirus family. To gain a better understanding of the viral and cellular events that occur upon initiation of infection, early in vitro transmission of HHV-8 to primary vascular endothelial cell populations was investigated. Materials and methods CellsBCBL-1 ...

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Section: Virus Preparation and Labelingmentioning

confidence: 70%

Characterization of Kaposi sarcoma–associated herpesvirus/human herpesvirus–8 infection of human vascular endothelial cells: early events

Dezube

Zambela

Sage

et al. 2002

Blood

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“…The increased yield of this virus would be highly advantageous for growth of stocks of an HSV-2 vaccine candidate, because the growth kinetics and yields of HSV-2 can be limiting in GMP production [57, 58]. The increased growth of the recombinant with an HSV-1 vhs gene, as compared with the parental strain containing the HSV-2 vhs, argues that the strong HSV-2 vhs activity may actually limit viral yields, while the increased yields relative to dl 5-29-41, which expresses no vhs protein, argues that either some vhs activity is helpful to viral growth or that the U L 41 protein enhances the formation or stability of viral particles.…”

Section: Discussionmentioning

confidence: 99%

Construction and properties of a herpes simplex virus 2 dl5-29 vaccine candidate strain encoding an HSV-1 virion host shutoff protein

Reszka

Dudek

Knipe

2010

Vaccine

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The replication-defective herpes simplex virus 2 (HSV-2) dl5-29 mutant virus strain with deletions in the UL5 and UL29 genes has been shown to protect mice and guinea pigs against challenge with wild type (wt) HSV-2 and to protect against ocular disease caused by HSV-1 infection. The dl5-29 strain is currently being prepared for clinical trials as a herpes vaccine candidate. As a possible approach to improve the efficacy of dl5-29 as a genital herpes vaccine, we replaced the UL41 gene encoding the virion host shutoff function (vhs) with the UL41 gene from HSV-1. While the HSV-2 UL41 and HSV-1 UL41 gene products have analogous functions, vhs-1 is 40-fold less active than vhs-2. Previously, it was shown that disruption of the UL41 gene can increase the efficacy of dl5-29 as a vaccine against HSV-2. These properties led us to hypothesize that replacement of vhs-2 by vhs-1 would decrease cytopathic effects in infected host cells, allowing longer survival of antigen-presenting cells and induction of stronger immune responses. The new recombinant dl5-29-41.1 virus shows nearly the same immunogenicity and protection against HSV-2 challenge as the parental dl5-29 virus or a triply deleted mutant virus, dl5-29-41, in the murine model of infection, and grows to higher titers than the parental strain in complementing cells, which is important for GMP production. The results have implications for the design of future HSV-2 vaccine candidates and mechanisms of induction of protective immunity against genital herpes.

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“…Indeed, such binding is important at the beginning of infection for attachment of incoming virions. Supporting a role for these interactions in release, it has been shown that polyanionic mimics of heparan sulfate can elute HSV-2 progeny virions from the surface of infected cells (64). It is also possible that progeny virions use the viral envelope protein gD to interact with cellular receptors, such as HVEM or nectin-1 (63).…”

Section: Discussionmentioning

confidence: 99%

Functional Comparison of Herpes Simplex Virus 1 (HSV-1) and HSV-2 ICP27 Homologs Reveals a Role for ICP27 in Virion Release

Park

Lalli

Sedlackova-Slavikova

et al. 2015

J Virol

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Numerous studies have focused on the regulatory functions of ICP27, an immediate-early (IE) protein of herpes simplex virus 1 (HSV-1). However, its homolog in HSV-2, termed ICP27t2, has been little studied. Here, we used two different approaches to functionally compare ICP27t2 and ICP27. In transfection-based assays, ICP27t2 closely resembled ICP27 in its capacity to enhance HSV-1 late gene expression, suppress the splicing of a viral intron, and complement the growth of an HSV-1 ICP27 null mutant. To study ICP27t2 in the context of viral infection, we engineered K2F1, an HSV-1 mutant that encodes ICP27t2 in place of ICP27. In Vero cells, K2F1 replicated with wild-type (WT) kinetics and yields, expressed delayed-early and late proteins normally, and was fully capable of activating several cellular signal transduction pathways that are ICP27 dependent. Thus, we conclude that ICP27t2 and ICP27 are functionally very similar and that ICP27t2 can mediate all ICP27 activities that are required for HSV-1 replication in cell culture. Surprisingly, however, we found that K2F1 forms plaques that are morphologically different from those of WT HSV-1. Investigation of this trait demonstrated that it results from the decreased release of progeny virions into the culture medium. This appears to be due to a reduction in the detachment of K2F1 progeny from the extracellular surface of the infected cell. We identified two HSV-1 ICP27 amino-terminal deletion mutants with a similar release defect. Together, these results demonstrate that ICP27 plays a heretofore-unappreciated role in modulating the efficiency of progeny virion release. IMPORTANCEICP27 is an essential, multifunctional regulatory protein that has a number of critical roles in the HSV-1 life cycle. Although ICP27 homologs are encoded by all known members of the Herpesviridae, previous work with several of these homologs has shown that they cannot substitute for ICP27 in the context of HSV-1-infected cells. Here, we identify ICP27t2 as the first homolog that can efficiently replace ICP27 in HSV-1 infection. Unexpectedly, our results also reveal that the sequence of the ICP27 gene can affect the release of HSV-1 progeny virions from the infected cell. Thus, our comparative study has revealed a novel function for ICP27 in the regulation of virus release. Herpes simplex virus 1 (HSV-1), a human alphaherpesvirus, is widespread in the human population, with greater than 57% of U.S. adults seropositive (1). The interaction of HSV-1 with the human host has been well studied (reviewed in reference 2). The virus is transmitted to new hosts via body secretions such as saliva, with the primary site of infection often being the oral mucosa. Following replication in epithelial cells, HSV-1 infects sensory neurons, where it establishes a lifelong latent infection in which the viral genome persists in the trigeminal ganglion as a semiquiescent episome. Periodically, the virus reactivates and returns to the oral cavity, where it can cause lesions (cold sores) and be transmitte...

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The primary production of an infectious recombinant Herpes Simplex Virus vaccine

Cited by 19 publications

References 21 publications

Characterization of Kaposi sarcoma–associated herpesvirus/human herpesvirus–8 infection of human vascular endothelial cells: early events

Characterization of Kaposi sarcoma–associated herpesvirus/human herpesvirus–8 infection of human vascular endothelial cells: early events

Construction and properties of a herpes simplex virus 2 dl5-29 vaccine candidate strain encoding an HSV-1 virion host shutoff protein

Functional Comparison of Herpes Simplex Virus 1 (HSV-1) and HSV-2 ICP27 Homologs Reveals a Role for ICP27 in Virion Release

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