herpes
Enhanced atherogenesis is not an obligatory response to systemic herpes virus infection in the apoE-deficient mouse: comparison of murine gamma-herpes virus-68 and herpes simplex virus-1.

Alber DG, Vallance P, Powell KL.

Wolfson Institute for Biomedical Research, University College London, London, UK. d.alber ucl.ac.uk

Viral and bacterial infectious agents have been implicated in the etiology of atherosclerosis. We have previously shown that a gamma-herpes virus can accelerate atherosclerosis in the apolipoprotein E-deficient (apoE-/-) mouse. To address whether a virally induced systemic immune response is sufficient to trigger enhanced atheroma formation, we infected apoE-/- mice with murine gamma-herpes virus-68 (MHV-68) or herpes simplex virus-1 (HSV-1). In this study, we show that both viruses were able to induce a cell-mediated and humoral immune response in the apoE-/- mouse, which was sustained over a period of 24 weeks. Although intranasal or intraperitoneal infection with MHV-68 induced similar levels of virus-specific IgG1 and IgG2a antibodies in the serum of apoE-/- mice, those infected with HSV-1 showed higher anti-HSV-1 IgG2a compared with IgG1 antibody levels. In addition, viral message was not detected in the aortas of HSV-1-infected animals, whereas we have shown previously that MHV-68 mRNA can be detected in the aortas of infected mice as early as 5 days after infection. Compared with control mice, apoE-/- mice infected with MHV-68 showed accelerated atherosclerosis, whereas mice infected with HSV-1 did not. These data indicate that a systemic immune response to any particular infectious agent is insufficient to induce enhanced atherosclerosis in the apoE-/- mouse and point to specific infections or immune mechanisms that might be essential for virally enhanced atherogenesis.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12006392&dopt=Abstract herpes medicine



herpes
[Development of a method for creating a colony of Macaca mulatta, free of herpes B virus infection]

[Article in Russian]

Klots IN, Rezikian SA, Indzhiia LV, Chalian VG, Meishvili NV, Lapin BA.

Institute of Medical Primatology, Sochi-Adler, Russia.

To form the colony of monkeys, free of herpes B virus, the serological study was made with the aim of finding out the carriers of this virus. 482 rhesus macaques (Macaca mulatta) from the Adler monkey house were examined for the presence of antibodies to herpes B virus by the method of point immunoblotting with the use of Herpes virus simiae as antigen. The contamination of monkeys in different open-air cages varied from 12.5% to 92%. In different age groups, it was 27% in nonpubescent monkeys (49 out of 182 animals), 55% in adolescent monkeys (55 out of 99), 73% in pubescent monkeys (131 out of 179) and 95% in monkeys over 15 years (21 out of 22 animals). 9 groups of rhesus macaques (comprising altogether 81 animals), free of herpes B virus, were selected. The monkeys were repeatedly tested within a year; after that 10-17% of formerly seronegative monkeys were rejected and removed from the selected group. After the third testing 2.5% more of the animals were found to have seroconversion. The colony of rhesus macaques thus created exists at present. The animals are subjected to constant serological observation.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9700888&dopt=Abstract herpes medicine



herpes
Tumor necrosis factor-alpha response and herpes virus infection in Bell's palsy.

Larsson C, Bernstrom-Lundberg C, Edstrom S, Bergstrom T.

Department of Clinical Virology, Goteborg University, Sweden.

OBJECTIVES: To attempt early diagnosis of patients with Bell's palsy by detection of herpesviral DNA in body fluids, and to investigate whether tumor necrosis factor-alpha (TNF-alpha), a cytokine associated with demyelination, is involved in the inflammatory response in this disease. STUDY DESIGN: Eleven patients with acute facial palsy admitted within 1 week after onset of the disease were followed in a consecutive prospective study. METHODS: Antibodies reactive to herpes viruses were determined by enzyme-linked immunosorbent assay in serum samples from acute and convalescent (> 2-week interval) cases. Intrathecal antibody response was investigated by immunoblotting. Polymerase chain reaction amplification of herpesviral DNA was attempted from samples of serum, cerebrospinal fluid, tear fluid, and saliva TNF-alpha and its soluble receptors (types I and II) were assessed in serum and cerebrospinal fluid samples. RESULTS: Ten of the 11 patients demonstrated serologic evidence of herpesviral primary infection or reactivation, supporting the evidence that herpes viruses are the most prevalent etiologic agents in Bell's palsy. Despite this, DNA amplifications by polymerase chain reaction were negative for herpes viruses in the body fluids tested. TNF-alpha concentrations were significantly elevated in serum, as compared with controls. Only one patient had a remaining facial nerve dysfunction at follow-up after 3 months. CONCLUSION: The absence of herpes DNA in body fluids in the acute stage of serologically confirmed Bell's palsy suggests that viral replication is transient in cases with an early restoration of the facial nerve function. The elevated serum levels of TNF-alpha indicate that this cytokine might be a pathogenetic factor related to the demyelination in this disease.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9707238&dopt=Abstract herpes medicine



herpes
Coagulation initiated on herpes viruses.

Sutherland MR, Raynor CM, Leenknegt H, Wright JF, Pryzdial EL.

The Canadian Red Cross Society, Research and Development Department, 1800 Alta Vista Drive, and Department of Biochemistry, University of Ottawa, Ottawa, ON Canada K1G 4J5.

Herpesviruses have been previously correlated to vascular disease and shown to cause thrombogenic and atherogenic changes to host cells. Herein we show that even in the absence of cells, purified cytomegalovirus (CMV) and herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) can initiate thrombin production. Functional assays demonstrated that purified HSV-1 and HSV-2 provide the necessary phospholipid (proPL) for assembling the coagulation factors Xa and Va into prothrombinase, which is responsible for generating thrombin. These observations are consistent with our earlier studies involving CMV. The presence of proPL on all three herpes viruses was confirmed directly by flow cytometry and electron microscopy by using annexin V and factor Va, respectively, as proPL-specific probes. Of equal importance, we found that CMV, HSV-1, and HSV-2 were also able to facilitate factor Xa generation from the inactive precursor factor X, but only when factor VII/VIIa and Ca2+ were present. Monoclonal antibodies specific for tissue factor (TF), the coagulation initiator, inhibited this factor X activation and, furthermore, enabled identification of TF antigen on each virus type by flow cytometry and electron microscopy. Collectively, these data show that CMV, HSV-1, and HSV-2 can initiate the generation of thrombin by having essential proPL and TF activities on their surface. Unlike the normal cellular source, the viral activity is constitutive and, therefore, not restricted to sites of vascular injury. Thus cell-independent thrombin production may be the earliest event in vascular pathology mediated by herpes viruses.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9391056&dopt=Abstract herpes medicine



herpes
Herpes simplex virus type-2, cytomegalovirus and Epstein-Barr virus infection in acute non A to E hepatitis Thai patients.

Maneerat Y, Wilairatana P, Pongponratn E, Puthavathana P, Chaisri U, Kurathong S, Juthaputhi A, Clayson ET, Snitbhan R, Raengsakulrach B, Vaughn DW.

Faculty of Tropical Medicine, Siriraj Hospital, Bangkok, Thailand.

A significant number of acute non A to E hepatitis cases are reported in Thailand every year, and the etiologies of these cases are unknown. Members of the herpesviridae family have been reported to cause either a self limited or fatal hepatitis in a small proportion of patients in other parts of the world. To determine whether herpes viruses may play a role in acute non A to E hepatitis, sera from 32 acute hepatitis patients without markers for acute hepatitis A to E virus infection were examined for IgM to herpes virus type 2 (HSV-2), cytomegalovirus (CMV), and Epstein-Barr virus (EBV) using commercially available assays. IgM to HSV-2 was detected in four sera, IgM to CMV was detected in one serum, and IgM to EBV was detected in one serum. All of the acute non A to E hepatitis patients recovered and none had underlying conditions associated with impaired immunity. These results suggest that herpes viruses should be considered in the differential diagnosis for Thai patients with hepatitis.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9438547&dopt=Abstract herpes medicine



herpes
Intracellular metabolism of the N7-substituted acyclic nucleoside analog 2-amino-7-(1,3-dihydroxy-2-propoxymethyl)purine, a potent inhibitor of herpes virus replication.

Neyts J, Balzarini J, Andrei G, Chaoyong Z, Snoeck R, Zimmermann A, Mertens T, Karlsson A, De Clercq E.

Rega Institute for Medical Research, Katholieke Universiteit Leuven, Belgium. johan.neyts rega.kuleuven.ac.be

We investigated the intracellular metabolism of S2242 (2-amino-7-(1,3-dihydroxy-2-propoxymethyl)purine), the only known antivirally active acyclic nucleoside analogue with the side chain substituted at the N7 position of the purine ring. Uptake of S2242 by CEM cells increased linearly with increasing extracellular concentrations of the compound and was blocked by inhibitors of nucleoside transport. S2242 was phosphorylated in a time- and concentration-dependent manner to its monophosphates, diphosphates, and triphosphates. Intracellular half-life of the diphosphates and triphosphates in CEM cells was approximately 3-6 hr. A strong correlation was found between the cytostatic action of the compound and its phosphorylation in different cell lines. In accord with the findings that (1) the cytostatic potential of S2242 is reversed by deoxycytidine (dCyd) and (2) the growth of deoxycytidine kinase-deficient (dCK-) cells is refractory to the inhibitory effect of S2242, the amount of metabolites formed from S2242 in the dCK- cell line was approximately one hundredth of that in the wild-type cells. The observation that purified dCK phosphorylates S2242 to its monophosphate further corroborates these results. The activity of S2242 against herpes simplex virus, varicella-zoster virus, and human herpes virus type 6 was reversed by 50-100-fold on the addition of exogenous dCyd. Compound S2242 was not preferentially phosphorylated in herpes simplex virus 1-, varicella-zoster virus-, or human herpes virus type 6-infected cells (Vero, human embryonic lung, and HSB-2 cells, respectively), and exogenously added dCyd reduced substantially the formation of S2242 metabolites in these cells. In human cytomegalovirus (HCMV)-infected human embryonic lung cells, a 5-25-fold increase in S2242 metabolite formation was observed compared with the noninfected cells, suggesting that an HCMV-encoded or -induced enzyme causes the specific phosphorylation of S2242. Exogenously added dCyd had little effect on the activity of S2242 against HCMV and on the phosphorylation of the compound in HCMV-infected cells. S2242 was not specifically phosphorylated by the HCMV-encoded UL-97 kinase in cells infected with a vaccinia/UL-97 recombinant. S2242 was found to be a substrate (K(m) = 90 microM) for purified human deoxyguanosine kinase; the latter enzyme was stimulated 3-4-fold in HCMV-infected cells.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9443944&dopt=Abstract herpes medicine



herpes
The bovine herpes virus type 1 UL3.5 open reading frame encodes a virion structural protein.

Schikora B, Lu Z, Kutish GF, Rock D, Magyar G, Letchworth GJ.

Department of Animal Health and Biomedical Sciences, University of Wisconsin, Madison 53706, USA.

The bovine herpes virus type 1 (BHV-1) open reading frame (ORF) UL3.5 is similar to ORFs found in pseudorabies virus, infectious laryngotracheitis virus, equine herpes virus type 1, and varicella zoster virus, but clearly absent from herpes simplex virus. The published sequence for this ORF predicts a 126-amino-acid (13.2 kDa) protein product with an isoelectric point of 12.3. We confirmed the UL3.5 sequence, expressed the ORF as a glutathione-S-transferase fusion protein, and made rabbit antibodies against the purified fusion protein. The antiserum detected a 13-kDa protein in Western blots of MDBK cells infected with BHV-1, but not with other herpes viruses or uninfected cells. The BHV-1 UL3.5 protein was characterized as a component of the virion envelope or tegument because it was expressed as a late protein, it was present in the cytoplasm but not the nucleus of infected cells, and it was removed from purified virions by detergent extraction.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9448691&dopt=Abstract herpes medicine



herpes
Simultaneous identification of two populations of sympathetic preganglionic neurons using recombinant herpes simplex virus type 1 expressing different reporter genes.

Levatte MA, Mabon PJ, Weaver LC, Dekaban GA.

Neurodegeneration Group, The John P. Robarts Research Institute, London, Ontario, Canada.

We generated neurotropic herpes simplex type 1 viruses expressing human placental alkaline phosphatase and studied the utility of this enzyme as a marker of infected neurons. The neurotropism of these viruses was assessed by their ability to infect sympathetic preganglionic neurons after adrenal injection in hamsters. The transneuronal transfer of these viruses was examined by their ability to cross the peripheral synapse from the kidney to renal preganglionic neurons or to cross the central synapse from the adrenal gland to the medulla oblongata. Finally, we injected an alkaline phosphatase-expressing herpes simplex virus into the adrenal gland and a beta-galactosidase-expressing herpes simplex virus (US5gal) into the muscular wall of the small intestine to label two neural circuits in one animal and to assess the feasibility of a dual-virus labelling system. The alkaline phosphatase gene was inserted into the glycoprotein J locus or the virus-induced host shut-off locus in the herpes simplex genome to create viruses which replicate (gJHAP HSV or vhsHAP HSV) or into the thymidine kinase locus to generate a virus that does not replicate in neurons in vivo (TK- HAP HSV). Each of the three viruses was retrogradely transported from the adrenal gland of hamsters to sympathetic preganglionic neurons, suggesting that the neurotropism of these viruses was maintained. gJHAP HSV travelled transneuronally from the kidney to sympathorenal preganglionic neurons and from the adrenal gland to neurons in the rostral ventrolateral medulla. Neuronal infection with alkaline phosphatase-expressing virus could be identified using histochemistry but detailed morphology of these neurons was not revealed. However, staining by anti-herpes simplex virus immunoperoxidase demonstrated that they had normal morphology. Identification of two distinct neural circuits in one animal was achieved with our dual-virus labelling system. The nonreplicating TK- HAP HSV was used in combination with US5gal to identify intestinal and adrenal sympathetic preganglionic neurons. The beta-galactosidase-expressing intestinal neurons were labelled bilaterally in the nucleus intermediolateralis, pars principalis, and alkaline phosphatase-expressing adrenal neurons were found ipsilaterally. Some clusters of sympathetic preganglionic neurons in the nucleus intermediolateralis, pars principalis contained mostly intestinal sympathetic preganglionic neurons and a few adrenal sympathetic preganglionic neurons. In other areas, the opposite pattern occurred. About 3-7% of the labelled sympathetic preganglionic neurons were double-labelled by both markers. The distinct and crisp morphology and dendritic processes of neurons stained by beta-galactosidase histochemistry contrasted with the partial staining of neurons by alkaline phosphatase, revealing beta-galactosidase as a better marker of infected neurons. In conclusion, alkaline phosphatase-expressing herpes simplex viruses are yet neurotropic after insertion of this marker enzyme into any of three different loci of the herpes simplex genome. One replicating alkaline phosphatase-expressing virus travelled transneuronally. These alkaline phosphatase-expressing herpes simplex virus can be used together with beta-galactosidase-expressing herpes simplex viruses to determine the target specificity of sympathetic preganglionic neurons controlling visceral organs or can be used to express two different recombinant genes in two targeted neuronal populations. This study suggests that sympathetic preganglionic neurons controlling the intestine and adrenal gland are almost completely distinct.

Online source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=9466444&dopt=Abstract herpes medicine