herpes
Cloning and functional analysis of Kaposi's sarcoma-associated herpes virus DNA polymerase and its processivity factor.

Lin K, Dai CY, Ricciardi RP.

Department of Microbiology, School of Dental Medicine, School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.

Kaposi's sarcoma-associated herpes virus (KSHV), or human herpes virus 8, is a newly identified virus with tumorigenic potential. Here, we cloned and expressed the DNA polymerase (Pol-8) of KSHV and its processivity factor (PF-8). Pol-8 bound specifically to PF-8 in vitro. Moreover, the DNA synthesis activity of Pol-8 was shown in vitro to be strongly dependent on PF-8. Addition of PF-8 to Pol-8 allowed efficient synthesis of fully extended DNA products corresponding to the full-length M13 template (7,249 nucleotides), whereas Pol-8 alone could incorporate only several nucleotides. The specificity of PF-8 and Pol-8 for each other was demonstrated by their inability to be functionally replaced by the DNA polymerases and processivity factors of herpes simplex virus 1 and human herpes virus 6.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9621095&dopt=Abstract herpes medicine



herpes
Prevalence of herpes simplex virus infection in patients suspected of genital herpes; and virus typing by type specific fluorescent monoclonal antibodies.

Puthavathana P, Kanyok R, Horthongkham N, Roongpisuthipong A.

Department of Microbiology, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand.

During the period between April 1994 and February 1996, a total of 154 female patients who attended the Clinic of Female Sexually Transmitted Diseases, Siriraj Hospital with clinical symptoms suspected of genital herpes were investigated for herpes simplex virus (HSV) infection by the virus isolation method in Vero cell cultures. Swabs from external genital lesions and the cervix from each patient were collected separately and used as the clinical specimens for isolation of HSV. The virus isolates were identified by indirect immunofluorescence (IIF) staining of the infected cell cultures using polyclonal HSV-2 specific antiserum which was reactive to common HSV antigens for both types of viruses. Typing of HSV was performed by direct IF using monoclonal antibody specific to HSV-1 or HSV-2. HSV was isolated from 78.6 per cent (121 of 154) of the cases studied; and among the infected cases, there were 47.9 per cent (58 of 121) in whom the infection involved both external genital lesions and cervixes, and 50.4 per cent (61) in whom the infection was limited to external genital lesions only. There were 2 cases (1.7%) in whom HSV was isolated from cervixes but not external genital lesions. Seventy-five HSV isolates were further subjected to typing. The present study showed that HSV-1 was accounted for 18.7 per cent (14 isolates), while HSV-2 took the remaining part of 81.3 per cent (61 isolates). The data demonstrated an increase in the prevalence of HSV-1 in genital herpes in our people.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9623019&dopt=Abstract herpes medicine



herpes
A cell surface protein with herpes virus entry activity (HveB) confers susceptibility to infection by mutants of herpes simplex virus type 1, herpes simplex virus type 2, and pseudorabies virus.

Warner MS, Geraghty RJ, Martinez WM, Montgomery RI, Whitbeck JC, Xu R, Eisenberg RJ, Cohen GH, Spear PG.

Department of Microbiology-Immunology, Northwestern University Medical School, Chicago, Illinois 60611, USA.

Certain mutant strains of herpes simplex virus type 1 (HSV-1) are unable to infect cells in which entry is dependent on HVEM, the previously described herpes virus entry mediator designated here as herpes virus entry protein A (HveA). These mutant viruses can infect other cells where entry is apparently dependent on other co-receptors. The mutant virus HSV-1(KOS)Rid1 was used to screen a human cDNA expression library for ability of transfected plasmids to convert resistant Chinese hamster ovary cells to susceptibility to virus entry. A plasmid expressing the previously described poliovirus receptor-related protein 2 (Prr2) was isolated on the basis of this activity. This protein, designated here as HveB, was shown to mediate the entry of three mutant HSV-1 strains that cannot use HVEM as co-receptor, but not wild-type HSV-1 strains. HveB also mediated the entry of HSV-2 and pseudorabies virus but not bovine herpes virus type 1. HveB was expressed in some human neuronal cell lines, fibroblastic cells, keratinocytes, and primary activated T lymphocytes. Antibodies specific for HveB blocked infection of HveB-expressing CHO cells and a human fibroblastic cell strain HEL299. Differences in ability of HSV-1 and HSV-2 strains to use HveB for entry should influence the types of cells that can be infected and thereby account in part for serotype and strain differences in tissue tropism and pathogenicity.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9657005&dopt=Abstract herpes medicine



herpes
The effect of gramicidin, a topical contraceptive and antimicrobial agent with anti-HIV activity, against herpes simplex viruses type 1 and 2 in vitro.

Bourinbaiar AS, Coleman CF.

Metatron, Inc., New York, New York, USA.

The effect of an anti-HIV compound, gramicidin, previously used as a topical antibiotic and vaginal contraceptive, on the replication of herpes simplex viruses (HSV) type 1 and 2 has been examined. Human WI-38 fibroblasts were inoculated with either HSV type in the presence of serial dilutions of gramicidin and reduction in viral yield was measured by ELISA. The 50% inhibitory dose (IC50) of gramicidin against 3 HSV-1 and 4 HSV-2 isolates was equal to 0.3 microgram/ml and was comparable to the efficacy of the anti-HSV agent acyclovir (ACV). The IC50 of gramicidin required to protect WI-38 from cytolytic effect of HSV was 10 micrograms/ml at day 5 postinfection, indicating that at this time point the activity of gramicidin was inferior than that of ACV. Nevertheless, gramicidin suppressed the replication of ACV-resistant thymidine kinase and DNA polymerase HSV mutants at doses effective against ACV-sensitive strains. The results suggest that the antimicrobial and spermostatic agent, gramicidin, has potential against sexually transmitted diseases (STDs) and for prophylaxis of sex-borne HIV and HSV infections.

PIP: Symptomatic human herpes simplex virus type 1 (HSV-1) infections are rather benign in immunocompetent individuals. The primary clinical manifestations of HSV-2 infection, which is mainly transmitted sexually, are anogenital lesions. Genital herpes affects one third of the world's population, and possibly 80% of those infected with HIV. HSV infections are especially severe and even life-threatening in people with AIDS. Only 20% of herpes seropositive persons have symptomatic infection, with the remainder asymptomatic but able to shed the virus. HSV infections are usually treated with nucleoside analogs such as acyclovir (ACV), but HSV eventually becomes resistant to ACV due to the loss or mutation of the viral thymidine kinase (TK) or changes in viral DNA polymerase. Gramicidin has recently been identified as a potent nontoxic anti-HIV agent 3-5 times more active than nonoxynol-9. Findings are reported from an assessment of the effect of gramicidin upon the replication of HSV-1 and HSV-2. Human WI-38 fibroblasts were inoculated with either HSV type in the presence of serial dilutions of gramicidin, while reduction in viral yield was measured by ELISA. The 50% inhibitory dose (IC50) of gramicidin against 3 HSV-1 and 4 HSV-2 isolates was equal to 0.3 mcg/ml and was comparable to the efficacy of ACV. The IC50 of gramicidin required to protect WI-38 from the cytolytic effect of HSV was 10 mcg/ml at day 5 postinfection, indicating that gramicidin was less active than ACV. Gramicidin nonetheless suppressed the replication of ACV-resistant thymidine kinase and DNA polymerase HSV mutants at doses effective against ACV-sensitive strains. These results suggest that gramicidin could be used against STDs and to prevent sexually transmitted HIV and HSV infections.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9672588&dopt=Abstract herpes medicine



herpes
Otarine herpes virus-1: a novel gammaherpes virus associated with urogenital carcinoma in California sea lions (Zalophus californianus).

King DP, Hure MC, Goldstein T, Aldridge BM, Gulland FM, Saliki JT, Buckles EL, Lowenstine LJ, Stott JL.

Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis, CA 95616, USA. dpking.vir gtnet.gov.uk

The incidence of neoplasia in California sea lions (CSLs) is considered to be unusually high. Electron microscopic examination of some of these urogenital tumours revealed the presence of virions with typical herpes-like structure. While current attempts to cultivate this virus have not been successful, molecular studies employing DNA extracted from tumour tissues allowed both the classification of the agent and its identification in tumours and archived tissue samples. Two genome fragments generated using degenerate primers in PCR demonstrated highest identities with other mammalian gammaherpes viruses. Phylogenetic analysis showed that this novel virus, tentatively designated Otarine herpes virus-1 (OtHV-1), grouped with members of the gammaherpes virus subfamily and was distinct from PHV-2, a previously described pinniped gammaherpes virus. An OtHV-1 specific PCR was established and used to investigate the presence of this virus in CSL tissues. PCR of DNA isolated from animals with these tumours, demonstrated that this virus was present in 100% (16/16) of tumours. Furthermore, DNA extracted from archived brain and muscle tissues was also positive in 29% (4/14) and 50% (7/14) of cases examined. This preliminary study provides evidence to support the hypothesis that the presence of this novel gammaherpes virus is a factor in the development of urogenital carcinoma in CSLs.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11888696&dopt=Abstract herpes medicine



herpes
Evaluation of specific herpes DNA viruses in idiopathic megarectum and idiopathic megacolon.

Gattuso JM, Debinski HS, Kangro HO, Jeffries D, Kamm MA.

St. Mark's Hospital, Harrow, Middlesex, UK.

PURPOSE: The aetiology of idiopathic megarectum and idiopathic megacolon is unknown. A previous study in patients with chronic idiopathic intestinal pseudo-obstruction, a condition also associated with a dilated gut, identified the possible involvement of herpes viruses. This study therefore aimed to determine whether these viruses may also be implicated in the pathogenesis of these conditions. METHODS: Resected large bowel from three patients with idiopathic megarectum and three patients with idiopathic megacolon were studied. Histology for viral inclusions and nested polymerase chain reaction (PCR) using specific primers for cytomegalovirus, Epstein-Barr virus, herpes simplex virus type 1 and varicella zoster virus was performed. DNA was extracted from paraffin-embedded blocks by proteinase K and phenol chloroform extraction. RESULTS: Viral inclusions were not seen. PCR failed to identify DNA of the four herpes viruses tested. CONCLUSION: Patients with idiopathic megarectum or idiopathic megacolon may have subtle abnormalities of the enteric innervation, but these do not appear to be attributable to the neurotropic effects of the herpes viruses studied.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9689563&dopt=Abstract herpes medicine



herpes
Treatment of recurrent genital herpes with interferon alpha-2alpha.

Cardamakis E, Relakis K, Kotoulas IG, Michopoulos J, Metallinos K, Mantouvalos H, Tzingounis V.

Department of Obstetrics and Gynecology, University of Patras, Diagnosis Diagnostic Center, Athens, Greece.

The purpose of the present study was to evaluate the efficacy and safety of parenteral administration of interferon alpha-2alpha in the treatment of recurrent herpes genitalis. A total of 97 patients (66 males, 31 females, mean age 34.86 +/-16.74 years), who had at least five recurrences of genital herpes during the previous 12 months, participated in a prospective open study on the effects of treatment with interferon alpha-2alpha (Roferon-A; Roche). The patients were treated with interferon alpha-2alpha (3 x 10(6) IU) by subcutaneous injection, three times weekly for 4 weeks, and the same schedule was repeated after 3 and 6 months. All patients were asymptomatic at the start of the study. After initiation of treatment, all patients reported to the clinic every 3 months for 2 years (the males were submitted to peoscopy and the females to Pap test and colposcopy) at the time of their recurrences. Comparison was made of the number of recurrences, duration of lesions, duration and severity of pain, and itching and burning. Prophylactic administration of interferon alpha-2alpha prevented recurrences of genital herpes virus infection in 51 patients (20 males and 31 females). Interferon administration shortened the healing time from 8.5 days before treatment to 2.5 days after treatment (p < 0.001). There was a significant reduction in the number of recurrences during the study period, from 7.46 before treatment to 2.64 after treatment (p < 0.001). On the basis of the overall efficacy and adverse effects, this regimen may be of value in the routine treatment of recurrent herpes.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9692344&dopt=Abstract herpes medicine



herpes
Recognition of herpes simplex virus type 2 tegument proteins by CD4 T cells infiltrating human genital herpes lesions.

Koelle DM, Frank JM, Johnson ML, Kwok WW.

Department of Medicine, University of Washington, Seattle, Washington 98105, USA.

The local cellular immune response to herpes simplex virus (HSV) is important in the control of recurrent HSV infection. The antiviral functions of infiltrating CD4-bearing T cells may include cytotoxicity, inhibition of viral growth, lymphokine secretion, and support of humoral and CD8 responses. The antigens recognized by many HSV-specific CD4 T cells localizing to genital HSV-2 lesions are unknown. T cells recognizing antigens encoded within map units 0. 67 to 0.73 of HSV DNA are frequently recovered from herpetic lesions. Expression cloning with this region of DNA now shows that tegument protein VP22 and the viral dUTPase, encoded by genes UL49 and UL50, respectively, are T-cell antigens. Separate epitopes in VP22 were defined for T-cell clones from each of three patients. Reactivity with the tegument protein encoded by UL21 was identified for an additional patient. Three new epitopes were identified in VP16, a tegument protein associated with VP22. Some tegument-specific CD4 T-cell clones exhibited cytotoxic activity against HSV-infected cells. These results suggest that herpes simplex tegument proteins are processed for antigen presentation in vivo and are possible candidate compounds for herpes simplex vaccines.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9696844&dopt=Abstract herpes medicine