herpes
Development of the hot spot-combined PCR assay for detection of retroviral insertions into Marek's disease virus.

Borenshtein R, Davidson I.

Division of Avian and Fish Diseases, Kimron Veterinary Institute, Israel.

A two-step PCR, the Hot Spot-combined PCR assay, was developed for the identification and characterization of recombinant viruses in Marek's disease (herpes) and retrovirus co-infections. In the first PCR the herpes virus genomic fragment, that was recognized in previous studies as a hot spot site for retroviral integration was amplified [reviewed in Bronovskis, P., Kung, H.-J., 1996. Retrotransposition and herpes virus evolution. Virus Genes 11, 259-270]. The products served for a second amplification step, performed in six PCR sets, using the six possible combinations of the two herpes and the retrovirus primer sets. Development of the assay employed DNA of the recombinant virus, RM1, which was created by in vitro co-cultivation of Marek's disease and reticuloendotheliosis viruses [Isfort et al., 1992. Proc. Natl. Acad. Sci. 89, 991-995; Witter et al., 1997. Avian Dis. 41, 407-421]. As the retroviral insertion site and junction sequences were determined previously [Jones et al., 1996. J. Virol. 70, 2460-2467], RMI served in the present study as a test virus for the development of the new assay. It is shown now that the Hot Spot-combined PCR can detect the retroviral insert in RM1. the MDV integration site and the insert orientation. For confirmation the herpes and retrovirus chimeric PCR products were sequenced and the results were similar to those published previously [Jones et al., 1996. J. Virol. 70, 2460-2467]. This assay might be adopted in additional systems to detect foreign inserts at suspected genomic sites.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10894628&dopt=Abstract herpes medicine



herpes
On the control of late gene expression in Kaposi's sarcoma-associated herpes virus (human herpes virus-8).

Chang J, Ganem D.

Howard Hughes Medical Institute and Departments of Microbiology and Medicine, University of California, San Francisco, CA 94143-0414, USA.

Herpesvirus late genes require viral DNA replication for maximal expression. Although late gene expression appears to require DNA replication in cis in alphaherpes viruses, studies in Epstein-Barr virus (EBV) suggest that this cis-requirement might not pertain to the gammaherpes viruses. Based on these findings, a system was created to investigate the elements required for the regulation of Kaposi's sarcoma-associated herpes virus (KSHV; human herpes virus-8) late gene expression. The transcript of a classic late gene encoding the viral assembly protein was characterized and reporter genes driven by the assembly protein promoter region were constructed. Unlike the EBV case, expression of a reporter gene under the control of the assembly protein promoter did not display authentic regulation when removed from the context of the viral genome. Although reporter expression rose in cells displaying lytic replication, this expression was not diminished by specific inhibitors of viral DNA synthesis. Minimal core promoters were similarly unable to reproduce late gene regulation. These results suggest that proper KSHV late gene expression is likely to be dependent upon virus lytic replication in cis and indicate that the regulation of KSHV late genes more closely resembles that observed in herpes simplex virus than that described for EBV.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10900043&dopt=Abstract herpes medicine



herpes
[Herpetic superinfection of pemphigus: 6 cases]

[Article in French]

Zouhair K, el Ouazzani T, Azzouzi S, Sqalli S, Lakhdar H.

Service de Dermato-Venereologie, CHU Ibn Rochd, Casablanca, Maroc.

BACKGROUND: Herpes simplex infection should be searched for in patients who experience unexplained aggravation of acantholytic dermatosis. Epidermal alterations appear to play an important favoring role. PATIENTS AND METHODS: We report 6 cases of Herpes simplex infections in patients with pemphigus observed at the Ibn Rochd hospital dermatology unit, Casablanca, over a 3-year period (1995-1998). RESULTS: There were 4 men and 2 women, age range 27-76 years. Superficial pemphigus was observed in 5 cases and pemphigus vulgaris in 1. In all cases, histology showed cytopathogenic Herpes simplex infection. The Herpes simplex infection appeared when the patients were given general corticosteroid therapy at doses from 1 to 1.5 mg/kg/d for 15 days to 1 month. In two cases, the corticosteroids were associated with an immunosuppressor (azathioprine, cyclophosphamide). Cure was rapid with systemic aciclovir, achieved in 5 to 20 days. Recurrence was observed in 2 cases. DISCUSSION: Herpes simplex superinfection in patients with pemphigus is classically described but uncommonly observed. In our experience, it has become more frequent over recent years since, before 1990, we observed no cases. Its prevalence may be underestimated. The diagnosis of Herpes simplex superinfection is sometimes difficult and should be suggested in case of relapse or resistance to treatment. The Herpes virus simulates most cases of pemphigus relapse, with a preferential localization on the bullous lesions. The painful nature of the eruption is highly suggestive of Herpes simplex infection as is the presence of vesicles grouped in bouquets. Both type 1 and type 2 viruses are observed. Cure is rapidly achieved with aciclovir in 5 to 15 days. Spontaneous cure has been reported in the literature as well as recurrent herpetic infection. In two cases in the literature, the Herpes eruption was fatal due to severe herpetic hepatitis and disseminated intravascular coagulation. Herpes simplex infection must be rapidly recognized in pemphigus patients so curative aciclovir treatment can be given early.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10604008&dopt=Abstract herpes medicine



herpes
Detection of herpes virus DNA in peripheral blood mononuclear cells and skin lesions of patients with pemphigus by polymerase chain reaction.

Tufano MA, Baroni A, Buommino E, Ruocco E, Lombardi ML, Ruocco V.

Istituto di Microbiologia, Facolta di Medicina e Chirurgia, II Universita degli Studi di Napoli, Naples, Italy.

Pemphigus is an autoimmune disease where both endogenous (genetic) and exogenous (environmental) factors play a part. Viral infections, in particular herpes virus infections, have been identified as a possible triggering factor for pemphigus. In this study, using the polymerase chain reaction, we studied peripheral blood mononuclear cells (PBMC) and skin biopsies from patients with pemphigus, and in some of these were able to demonstrate the presence of DNA sequences of herpes simplex virus 1/2 (50% in PBMC and 71% in skin biopsies), Epstein-Barr virus (15% in PBMC and 5% in skin biopsies) and human herpes virus 6 (20% in PBMC only). However, the inability to detect herpes virus DNA consistently in these cases suggests that viral infection may only be an occasional factor triggering the outbreak or exacerbation of the disease. The possible role of interferons and interleukins in the pathogenesis of virus-induced pemphigus is discussed.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10606848&dopt=Abstract herpes medicine



herpes
Nectin2alpha (PRR2alpha or HveB) and nectin2delta are low-efficiency mediators for entry of herpes simplex virus mutants carrying the Leu25Pro substitution in glycoprotein D.

Lopez M, Cocchi F, Menotti L, Avitabile E, Dubreuil P, Campadelli-Fiume G.

Institute of Cancerology and Immunology, INSERM U119, Marseille, France.

The receptors for entry of herpes simplex viruses 1 and 2 (HSV-1 and -2), widely expressed in human cell lines, are members of a subset of the immunoglobulin superfamily exemplified by herpes virus entry mediator C (HveC) and the herpes virus immunoglobulin-like receptor (HIgR). This report focuses on two members of this subset, herpes virus entry mediator B (HveB), recently designated nectin2/PRR2alpha, and its splice variant isoform, nectin2/PRR2delta. Nectin2alpha and -delta share the ectodomain but differ in the transmembrane and cytoplasmic regions. HveB was reported to enable entry of HSV-1 carrying mutations in glycoprotein D (gD) and of HSV-2, but not of wild-type (wt) HSV-1. We report that (i) both nectin2alpha and -delta served as receptors for the entry of HSV-1 mutant viruses HSV-1(U10) and -(U21) and AP7(r) that carry the Leu25Pro substitution in gD but not for HSV-1 mutants U30 and R5000 that carry the Ser140 or Ala185 substitution in gD. All of these mutants were able to overcome the block to entry mediated by expression of wt gD. (ii) Infection of cells expressing nectin2alpha or -delta required exposure to multiplicities of infection about 100-fold higher than those required to infect cells expressing HveC or HIgR. (iii) gD from HSV-1(U21) bound in vitro soluble forms of nectin2. The association was weaker than that to the soluble form of HveC/HIgR. Binding of wt HSV-1 gD to soluble nectin2 was not detectable. (iv) A major region of nectin2 functional in virus entry mapped to the V domain, located at the N terminus.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10627537&dopt=Abstract herpes medicine



herpes
The ends on herpes virus DNA replicative concatemers contain pac2 cis cleavage/packaging elements and their formation is controlled by terminal cis sequences.

McVoy MA, Nixon DE, Hur JK, Adler SP.

Department of Pediatrics, Medical College of Virginia Campus of Virginia Commonwealth University, Richmond, Virginia 23298-0163, USA. mmcvoy hsc.vcu.edu

Herpesviruses have large double-stranded linear DNA genomes that are formed by site-specific cleavage from complex concatemeric intermediates. In this process, only one of the two genomic ends are formed on the concatemer. Although the mechanism underlying this asymmetry is not known, one explanation is that single genomes are cleaved off of concatemer ends in a preferred direction. This implies that cis elements control the direction of packaging. Two highly conserved cis elements named pac1 and pac2 lie near opposite ends of herpes virus genomes and are important for cleavage and packaging. By comparison of published reports and by analysis of two additional herpes viruses, we found that pac2 elements lie near the ends formed on replicative concatemers of four herpes viruses: herpes simplex virus type 1, equine herpes virus 1, guinea pig cytomegalovirus, and murine cytomegalovirus. Formation of pac2 ends on concatemers depended on terminal cis sequences, since ectopic cleavage sites engineered into the murine cytomegalovirus genome mediated formation of pac2 ends on concatemers regardless of the orientation of their insertion. These findings are consistent with a model in which pac2 elements at concatemer ends impart a directionality to concatemer packaging by binding proteins that initiate insertion of concatemer ends into empty capsids.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10627574&dopt=Abstract herpes medicine



herpes
Oncolytic herpes virus effectively treats murine squamous cell carcinoma and spreads by natural lymphatics to treat sites of lymphatic metastases.

Wong RJ, Joe JK, Kim SH, Shah JP, Horsburgh B, Fong Y.

Head and Neck Service, Department of Surgery, Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA.

Oncolytic herpes viruses have significant antitumoral effects in animal models when delivered directly to established tumors. Lymphatic metastases are a common occurrence for many tumor types. This study investigates the potential of an attenuated, replication-competent, oncolytic herpes simplex virus (NV1023) both to treat a primary tumor by direct injection and to travel through the lymphatic system to treat metastatic tumor within the lymph nodes draining lymph from the site of primary cancer. Isosulfan blue dye was injected into murine auricles to determine normal lymphatic drainage patterns and demonstrated consistent blue staining of a group of ipsilateral cervical lymph nodes. Auricular injections of NV1023 resulted in viral transit to these lymph nodes as measured by 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside histochemistry and viral plaque assay. An oncolytic herpes virus (NV1066) expressing green fluorescent protein also demonstrated viral transit from the auricle to the cervical lymph nodes on fluorescence microscopy. Using the SCC VII cell line, a novel murine model of auricular squamous cell carcinoma was developed with an approximately 20% incidence of cervical lymph node metastases. Delivery of NV1023 or NV1066 to the surgical beds after excision of auricular SCC VII tumors resulted in successful viral infection of metastatic SCC VII cells within the cervical lymph nodes. After a 7-week follow-up, significantly enhanced locoregional control (p < 0.05, Fisher exact test) and disease-free survival (p < 0.05, log rank test) were evident with NV1023 treatment. This study demonstrates that the delivery of an oncolytic herpes virus to a primary tumor site after surgical excision may have a significant impact on reducing both primary site recurrence and regional nodal metastases.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12133274&dopt=Abstract herpes medicine



herpes
[Recurrent eruptions of herpes zoster]

[Article in Czech]

Cerny Z.

Department of Infectious Diseases, Medical Faculty, Masaryk University, Brno, Czech Republic. zcerny med.muni.cz

BACKGROUND: Repeated eruptions of herpes zoster are frequently detected in immunocompromised patients, but rarely may occur also in immunocompetent persons. While detailed analyses of repeated manifestations of this disease in the literature are scarce, experience with a small group of affected persons is presented. AIMS OF THE STUDY: To search for the nature of herpes zoster relapses occurrence and to use it in treatment quality improvement. METHODS: Analysis and processing of clinical documentation of the first and repeated eruptions of herpes zoster in 12 patients. Study of sex and age, presence of accompanying diseases, duration of the interval from the first eruption to the relapse, localization, complications and relation to antiviral therapy. RESULTS: The interval between the first and repeated manifestation ranged from 1 week to 30 years. The male to female ratio was 10/2. In 3 patients the disease was accompanied by malignancy, in 3 by diabetes mellitus and in 1 by proved immunodeficiency. The most frequent eruption localization was intercostal space. In 6 patients the relapse was localized at the same site, in 6 of them at different. Early relapses in the same localization appeared in immunodeficient persons and also in persons after the first attack treatment with acyklovir. Primary manifestation was accompanied by complications in 42%, repeated in 92%. CONCLUSIONS: Early herpes zoster relapses following shortly after the primary attack treated with acyklovir should be considered as a manifestation of immunity disorder requiring immediate treatment with other antiviral drug. Late relapses can be treated with acyclovir. (Tab. 2, Fig. 2, Ref. 22.)

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10645043&dopt=Abstract herpes medicine