Bovine herpes simplex virus 1 glycoprotein B does not productively interact with cell surface heparan sulfate in a pseudorabies virion background.

Attachment to cell surface heparan sulfate proteoglycans is the first step in infection by several alphaherpes viruses. This interaction is primarily mediated by virion glycoprotein C (gC). In herpes simplex virus, in the absence of the nonessential gC, heparan sulfate binding is effected by glycoprotein B. In contrast, gC-negative pseudorabies virus (PrV) infects target cells via a heparan sulfate-independent mechanism, indicating that PrV virion gB does not productively interact with heparan sulfate. To assay whether a heterologous alphaherpes virus gB protein will confer productive heparan sulfate binding on gC-negative PrV, gC was deleted from an infectious PrV recombinant, PrV-9112C2, which expresses bovine herpes simplex virus 1 (BHV-1) gB instead of PrV gB. Our data show that gC-negative PrV-BHV-1 gB recombinant 9112C2-delta gCbeta was not inhibited in infection by soluble heparin, in contrast to the gC-positive parental strain. Similar results were obtained when wild-type BHV-1 was compared with a gC-negative BHV-1 mutant. Moreover, infection of cells proficient or deficient in heparan sulfate biosynthesis occurred with equal efficiency by PrV-9112C2-delta gCbeta, whereas heparan sulfate-positive cells showed an approximately fivefold higher plating efficiency than heparan sulfate-negative cells with the parental gC-positive virus. In summary, our data show that in a PrV gC-negative virion background, BHV-1 gB is not able to mediate infection by productive interaction with heparan sulfate, and they indicate the same lack of heparin interaction for BHV-1 gB in gC-negative BHV-1.

Production of herpes simplex B virus recombinant glycoproteins and evaluation of their diagnostic potential.

B virus (cercopithecine herpes simplex virus 1) is the only deadly alphaherpes virus that is zoonotically transmissible from macaques to humans. The detection of humoral immune responses is the method of choice for the rapid identification of B virus-infected animals. We evaluated the diagnostic potential of recombinant B virus glycoproteins for the detection of immunoglobulin G (IgG) antibodies in monkey and human sera. Glycoproteins B, C, and E and secreted (sgG) and membrane-associated (mgG) segments of glycoprotein G (gG) were expressed in the baculovirus expression system, while gD was expressed in CHO cells. We developed recombinant protein-based IgG enzyme-linked immunosorbent assays (ELISAs) and compared their diagnostic efficacies by using B virus antibody-negative (n = 40) and -positive (n = 75) macaque sera identified by a whole antigen-based ELISA and Western blotting. The diagnostic sensitivities of the gB-, gC-, gD-, and mgG-ELISAs were 100, 97.3, 88.0, and 80.0%, respectively. The specificities of the gB-, gC-, and gD-ELISAs and of the mgG-ELISA were 100 and 97.5%, respectively. In contrast, the sensitivities and specificities of sgG- and gE-ELISAs were low, suggesting that sgG and gE are less effective diagnostic antigens. Sera from nonmacaque monkeys cross-reacted with gB, gC, and gD, and only baboon sera reacted weakly with mgG. Human herpes simplex virus type 1 (HSV-1)- and HSV-2-positive sera pools reacted with gB and gD, whereas sera from B virus-infected individuals reacted with all four antigens. These data indicate that gB, gC, gD, and mgG have a high diagnostic potential for B virus serodiagnosis in macaques, whereas mgG may be a valuable antigen for discrimination between antibodies induced by B virus and those induced by other, closely related alphaherpes viruses, including HSV-1 and -2.

Cytomegalovirus periodontal presence is associated with subgingival Dialister pneumosintes and alveolar bone loss.

Destructive periodontal disease is associated with human cytomegalovirus (HCMV), Epstein-Barr type 1 virus (EBV-1) and other members of the Herpesviridae family as well as with various gram-negative anaerobic bacteria, including the Dialister pneumosintes species. This study aimed to determine possible interrelationships between periodontal HCMV, EBV-1, herpes simplex virus and D. pneumosintes, and relate the microbiological findings to periodontitis clinical status. Sixteen subjects each contributed paper point samples from two progressing and two stable periodontitis lesions, as determined by ongoing loss of probing attachment. Polymerase chain reaction methodology was used to identify the study herpes simplex viruses and D. pneumosintes. Chi-squared tests, Fisher exact tests and multivariate logistic regression were employed to identify statistical associations among herpes simplex viruses, bacteria and clinical variables. HCMV, and no other virus or combination of viruses, was positively associated with the presence of D. pneumosintes, and the relationship was specific for individual periodontitis sites with no detectable subject effect. D. pneumosintes was in turn positively associated with periodontal pocket depth and disease-active periodontitis. When the average percentage of alveolar bone loss in all teeth was treated as a response, HCMV remained significant even after D. pneumosintes was included in the model, suggesting that both HCMV and D. pneumosintes affected bone loss or, alternatively, HCMV affected factors not studied that themselves can induce bone loss. We hypothesize that periodontal HCMV sets the stage for subgingival proliferation of D. pneumosintes and subsequent periodontal disease progression. Studies on herpesviral-bacterial interactions may hold great promise for delineating important etio-pathogenic aspects of destructive periodontal disease.

Prevention of herpes simplex genitalis by the 'Bulgarian' vaccine F.HSV-2V(PRK): preliminary clinical evidence.

AIM: To examine the antigenic properties of the formalin-inactivated herpes simplex virus type 2 (HSV-2) virus-particle vaccine F. HSV-2V(PRK), which has been used therapeutically in Bulgaria for 30 years, and to make preliminary assessment of its potential protective efficacy by a follow-up of vaccinated patients with herpes simplex genitalis. METHODS: Properties of the vaccine were examined by standard immunological laboratory tests. Fifty-five patients at risk of herpes simplex genitalis received 2-4 vaccinations and were monitored during a 6-year follow-up. RESULTS: The vaccine was antigenic in laboratory tests and absorbed neutralizing antibody from hyperimmune rabbit serum against herpes simplex virus type 1 (HSV-1). In vaccinated patients, there was an overall contraction rate of herpes simplex genitalis of 5.4%. There was no evidence of significant local or generalized adverse effects from vaccination. CONCLUSION: Bulgarian vaccine F.HSV-2V(PRK) may have protective efficacy, which, in association with its apparent safety from our findings and from its clinical use for over 30 years in Bulgaria, suggests that it should be scrutinized by a formal clinical trial.

Herpes Simplex keratitis after ophthalmic surgery

PURPOSE: We report 6 cases of herpes simplex keratitis after ophthalmic surgery, in eyes without clinical history of herpes simplex keratitis. CASES: These cases comprised 6 patients examined at our hospital between April 1992 and November 2001. Past operations were keratoplasty in 5 eyes and cataract surgery in 1 eye. Clinical findings and predisposing factors were evaluated retrospectively. The period between herpetic epithelial keratitis onset and ophthalmic surgery ranged from 1.5 to 79 months. Predisposing factors included corticosteroid therapy and operative wound. The herpetic epithelial lesions were dendritic ulcers in 2 eyes, geographic ulcer in 1 eye, and atypical epithelial lesions in 3 eyes; in all cases, herpes simplex virus (HSV)-DNA was detected by polymerase chain reaction (PCR) in tear fluid. All herpetic epithelial lesions healed with oral and topical Acyclovir. CONCLUSIONS: When corticosteroids are used following ophthalmic surgery, physicians should be alert to the possibility of herpetic epithelial keratitis, even in patients with no clinical history of herpes simplex keratitis. PCR detection in tear fluid is helpful in diagnosing this disease.

Detection of herpes simplex virus infection of the CNS: the experience of hospital Geral de Santo Antonio.

BACKGROUND: PCR detection of CSF Herpes virus DNA is an important tool in the diagnosis of CNS infections. Use of this test has been shown to have an impact on patient management as measured by shortened patient stays, specific therapeutic intervention, reduction of empirical expensive therapy administration, all of which should translate into significant health care savings. OBJECTIVE: The present study aimed at implementing, and evaluating both clinically and analytically the performance of several commercially available PCR based assays for the detection of Herpes virus infections of the CNS. STUDY DESIGN: A total of 314 patients with suspected CNS Herpesvirus infection were investigated, between 1999 and 2001, by Stair primers PCR. Starting on January 2002, two commercially available real-time-PCR systems were implemented and tested using the Stair primers PCR assay as golden standard and three external control proficiency panels along with serial dilutions of positive clinical samples. RESULTS: Sensitivity of the assay was determined to be <200 copies per ml for HSV and <1250 copies per ml for CMV. Positive results were obtained for 17 patients (6 HSV-1, 1 HSV-2, 1 EBV, 1 CMV, 6 VZV and 2 HHV-6) whose clinical and analytical findings were consistent with the PCR results. A real-time-PCR procedure was introduced in 2002 with similar sensitivity, but a more rapid response. CONCLUSION: Conventional end-point PCR proved useful to the diagnosis of CNS herpes simplex virus infection, with an impact on the clinical intervention. However, the use of Real-Time-PCR has greatly enhanced these advantages, making results available at a much earlier time, thus significantly reducing the need for empirical treatment.

Prognostic value of Hutchinson's sign in acute herpes zoster ophthalmicus.

PURPOSE: To determine the prognostic value of nasociliary skin lesions (Hutchinson's sign) for ocular inflammation and corneal sensory denervation in acute herpes zoster ophthalmicus. METHODS: A longitudinal observational study with a 2-month follow-up was performed involving 83 non-immunocompromised adults with acute herpes zoster ophthalmicus, with a skin rash duration of less than 7 days, referred by their general practitioner. All skin lesions at the tip, the side and the root of the nose, representing the dermatomes of the external nasal and infratrochlear branches of the nasociliary nerve, were documented by taking photographs and marking anatomical drawings. Ocular inflammatory signs were observed by slit-lamp biomicroscopy, and corneal sensitivity was measured with the Cochet-Bonnet esthesiometer at 2-month follow-up. RESULTS: Hutchinson's sign was a powerful predictor of ocular inflammation and corneal denervation in herpes zoster ophthalmicus [relative risks: 3.35 (CI 95%: 1.82-6.15) and 4.02 (CI 95%:1.55-10.42), respectively]. The manifestation of herpes zoster skin lesions at the dermatomes of both nasociliary branches was invariably associated with the development of ocular inflammation. CONCLUSION: Clinicians should be alert for early skin lesions within the complete nasociliary dermatome, because they are a reliable prognostic sign of sight-threatening ocular complications in acute herpes zoster ophthalmicus.

A role for a new herpes simplex virus (KSHV) in different forms of Kaposi's sarcoma.

PIP: Samples were examined by polymerase chain reaction (PCR) for the presence of the putative Kaposi's sarcoma herpes simplex virus (KSHV). KS DNA from HIV-negative, African, endemic (EKS) samples, and epidemic HIV-positive KS (AKS), and sporadic KS (SKS) samples were tested from Tanzania and Sweden. All of the HIV KS (18 African EKS and 4 Swedish SKS) as well as the HIV-positive AIDS-related KS (16 African and 7 Swedish AKS) biopsies were shown to contain the previously described DNA sequences. KS lesions from children, females, and males in various tissues were analyzed including skin, lymph nodes, gut and oral mucosa. All forms of KS showed a single PCR product of the expected size (233 base pairs). To exclude amplification of other types of herpes simplex virus, virus preparations of Epstein-Barr virus (EBV), herpes simplex virus, cytomegalovirus, vesicular stomatitis, and human herpes simplex virus type 6 (HHV6) were assayed, again by PCR, using the KSHV primers. No PCR products were obtained with any of these virus strains. However, most HIV-positive and HIV-negative KS DNA samples also contained either EBV and/or HHV6 sequences. All biopsies from non-KS tissues (cells) of HIV-positive and HIV-negative individuals were consistently negative for KSHV by PCR. The observation that the same herpes simplex virus-like DNA sequence is present in endemic and sporadic, as well as AIDS-related, Kaposi's sarcoma cases suggests a possible pathogenic association between this putative novel, herpes-like virus and KS. The herpes simplex virus-like DNA sequences described by Y. Chang in 1994 may indeed represent a novel herpes simplex (KSHV), etiopathologically associated with various clinical forms of Kaposi's sarcoma. Its pathogenic importance is indicated by its presence in different KS tissues with various clinical types of KS and its absence from non-KS-involved tissues. Furthermore, the presence of KSHV in KS of children suggests a nonsexual mode of transmission.

Concatemeric intermediates of equine herpes simplex virus type 1 DNA replication contain frequent inversions of adjacent long segments of the viral genome.

In common with other alpha-herpes viruses, the genome of equine herpes simplex virus type-1 (EHV-1) comprises covalently linked long and short unique sequences of DNA, each flanked by inverted repeats. Equimolar amounts of two genomic isomers, generated by free inversion of the short segment, relative to the long segment, are packaged into EHV-1 virions. In contrast with herpes simplex virus (HSV), inversion of genomic long segments has not been described. In the current work, the structures of high molecular weight intermediates of EHV-1 DNA replication were studied by field inversion gel electrophoresis. It is shown that adjacent long segments of the viral genome are frequently inverted in concatemeric intermediates of EHV-1 DNA replication. Further, like HSV concatemers, high molecular weight intermediates of EHV-1 replication are flanked exclusively by the long segment of the viral genome. Hence, despite the fact that only two, rather than four, isomers of EHV-1 DNA are packaged into virions, the intermediates of EHV-1 DNA replication closely resemble those of herpes simplex virus type 1 in structure. These data have implications relating to the mechanisms involved in packaging of alpha-herpes virus DNA.

The potential role of suppressive therapy for sex partners in the prevention of neonatal herpes: a health economic analysis.

BACKGROUND: The development of suppressive therapy and type specific tests for herpes simplex infections allow for screening to reduce the risk of neonatal herpes. OBJECTIVES: To assess the potential effectiveness, cost effectiveness, and benefit of suppressive therapy among herpes simplex virus serodiscordant sex partners during pregnancy. METHODS: Decision and economic analyses are used to compare the incidence and costs of neonatal herpes simplex in California (2000) for three interventions: (1) no management; (2) current guidelines (caesarean delivery for women with lesions); (3) screening for women at risk and use of suppressive treatment in sex partners. RESULTS: Screening and suppressive therapy are the most effective interventions, while current guidelines have limited effectiveness, but the latter provide the most cost effective results. CONCLUSIONS: While current guidelines are cost saving, they forgo a potential 82% decrease in neonatal herpes simplex incidence that would be possible with screening and suppressive therapy if society were willing to pay the necessary US$363 000 per case prevented. To evaluate HSV screening and drug therapy completely, clinical trials and an economic assessment of infant mortality "value" to society are required.