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PubMed2002

Use of PCR for the diagnosis of herpesvirus infections of the central nervous system.

Polymerase chain reaction (PCR) analysis of cerebrospinal fluid (CSF) has revolutionized the diagnosis of nervous system viral infections, particularly those caused by human herpesviruses (HHV). The PCR technique allows the detection of minute quantities of DNA or RNA in body fluids and tissues. Both fresh-frozen and formalin-fixed tissues may be utilized for PCR assays, with the latter making archival studies possible. CSF PCR has now replaced brain biopsy as the gold standard for the diagnosis of herpes simplex virus (HSV) encephalitis. PCR analysis of both CSF and nervous system tissues has also broadened our understanding of the spectrum of disease caused by HSV-1 and -2, cytomegalovirus (CMV), Epstein-Barr virus (EBV), varicella zoster virus (VZV) and HHV-6. PCR results obtained from tissue specimens must be interpreted cautiously, since this highly sensitive technique may detect portions of viral genomic material that may be present even in the absence of active viral infection. Tissue PCR results in particular must be corroborated with clinical and neuropathologic evidence of central nervous system (CNS) infection. In several neurological diseases, negative PCR results have provided evidence against a role for herpesviruses as the causative agents. This review summarizes the role of CSF PCR in the diagnosis and therapeutic management of herpesvirus infections of the nervous system, particularly those caused by HSV and VZV.

Central Nervous System Viral Diseases↗
PubMed1987

Immunocytochemical confirmation of oral and respiratory herpesvirus infection by the immunoperoxidase method.

Recognition of diagnostic viral changes in general, and due to herpesvirus infection (HVI) in particular, from a variety of materials examined cytologically has added a new dimension to diagnostic cytology. Such recognition provides valuable information to the clinician when no infection is suspected. Previously, the diagnosis of HVI of the respiratory tract was only possible at postmortem, tissue culture, or diagnostic rise in antibody titer. This study describes our experience in the diagnosis of HVI of the respiratory tract initially suggested by sputum cytology and later confirmed by the immunoperoxidase method. The importance of sputum cytology in the diagnosis of HVI is emphasized.

Adult↗
PubMed1978

Therapeutic effect of trisodium phosphonoformate on cutaneous herpesvirus infection in guinea pigs.

When applied topically, trisodium phosphonoformate (PFA) displayed activity against established cutaneous herpesvirus infections in guinea pigs similar to that exhibited by the closely related phosphonoacetic acid (PAA); however, unlike PAA, PFA was not locally skin irritating. The therapeutic benefits of topical application of PFA were clearly evident when application was delayed for 48 h after virus inoculation, at which time lesions were well developed. The therapeutic effect was dependent on the concentration of PFA and the duration of treatment. PFA exhibited significant activity against established infections when administered intraperitoneally, although it was less effective via this systemic route than when applied topically.

Animals↗
PubMed1982

The effect of herpesvirus infection on ribosomal RNA synthesis and on nucleolar size and number in HeLa cells.

The effects of infection by the herpesvirus pseudorabies virus on the nucleolus and on ribosomal RNA synthesis in HeLa cells were examined. The size of individual nucleoli and the total nucleolar content per cell are reduced to approximately 35 per cent of normal by 6 hours of infection. The proportion of cells containing one nucleolus rises from 10 to 50 per cent in the same time. The synthesis of ribosomal precursor RNA declines to approximately 40 per cent in this period.

Cell Nucleolus↗
PubMed1987

Cyclophosphamide immunosuppression during lymphotropic herpesvirus infection in the guinea pig model. A histopathologic and virologic study.

Guinea pigs infected with lymphotropic herpesvirus (GPHLV) were given the immunosuppressive agent cyclophosphamide (Cy). All Cy-treated animals revealed the expected lymphoid depletion of spleen and lymph node B zones. Acute GPHLV infection of Cy-treated animals resulted in increased blood and spleen leukocyte viral infectivity titers and lymphoid tissue lesions containing cells positive for GPHLV antigen and intranuclear inclusions. During latent GPHLV infection, Cy treatment resulted in declining leukocyte viral infectivity titers without pathologic lesions. Morphologic data suggest that tissue histiocytic cells may be involved in the productive viral infection observed in Cy-immunosuppressed animals during acute GPHLV infection. During latency, however, infectious virus appears restricted to a Cy-sensitive, probably lymphoid, cell. This animal model appears useful for the study of lymphotropic viral infection during immunosuppression.

Animals↗
PubMed2005

Effect of herpesvirus infection on pancreatic duct cell secretion.

AIM: To examine the effect of acute infection caused by herpesvirus (pseudorabies virus, PRV) on pancreatic ductal secretion. METHODS: The virulent Ba-DupGreen (BDG) and non-virulent Ka-RREp0lacgfp (KEG) genetically modified strains of PRV were used in this study and both of them contain the gene for green fluorescent protein (GFP). Small intra/interlobular ducts were infected with BDG virus (10(7) PFU/mL for 6 h) or with KEG virus (10(10) PFU/mL for 6 h), while non-infected ducts were incubated only with the culture media. The ducts were then cultured for a further 18 h. The rate of HCO(3)(-) secretion (base efflux -J(B-)) was determined from the buffering capacity of the cells and the initial rate of intracellular acidification (1) after sudden blockage of basolateral base loaders with dihydro-4,4-diisothiocyanatostilbene-2,2-disulfonic acid (500 micromol/L) and amiloride (200 micromol/L), and (2) after alkali loading the ducts by exposure to NH(4)Cl. All the experiments were performed in HCO(3)(-)-buffered Ringer solution at 37 degrees (n = 5 ducts for each experimental condition). Viral structural proteins were visualized by immunohistochemistry. Virally-encoded GFP and immunofluorescence signals were recorded by a confocal laser scanning microscope. RESULTS: The BDG virus infected the majority of accessible cells of the duct as judged by the appearance of GFP and viral antigens in the ductal cells. KEG virus caused a similarly high efficiency of infection. After blockage of basolateral base loaders, BDG infection significantly elevated -J(B-) 24 h after the infection, compared to the non-infected group. However, KEG infection did not modify -J(B-). After alkali loading the ducts, -J(B-) was significantly elevated in the BDG group compared to the control group 24 h after the infection. As we found with the inhibitor stop method, no change was observed in the group KEG compared to the non-infected group. CONCLUSION: Incubation with the BDG or KEG strains of PRV results in an effective infection of ductal epithelial cells. The BDG strain of PRV, which is able to initiate a lytic viral cycle, stimulates HCO(3)(-) secretion in guinea pig pancreatic duct by about four- to fivefold, 24 h after the infection. However, the KEG strain of PRV, which can infect, but fails to replicate, has no effect on HCO(3)(-) secretion. We suggest that this response of pancreatic ducts to virulent PRV infection may represent a defense mechanism against invasive pathogens to avoid pancreatic injury.

Animals↗
PubMed1987

Laboratory diagnosis of herpesvirus infections.

The diagnosis of infection due to herpesviruses can be performed by several laboratory techniques, including virus culture, demonstration of virus-specific cellular antigens, detection of virus-induced serum antibodies, detection of viral nucleic acids (using radiolabeled probes), or visualization of histologic changes in infected tissue. Virus culture is the most sensitive technique for diagnosis of most infections due to herpes simplex virus (HSV) and cytomegalovirus (CMV), but is insensitive for the diagnosis of varicella-zoster (VZV) infection and not routinely available for Epstein-Barr virus (EBV). Identification of viral antigens on infected cells with monoclonal antibodies is useful for definitive identification of virus in cell culture and for rapid diagnosis of infection due to HSV, VZV, or CMV. Serologic studies are useful for the diagnosis of EBV infection, because clinical symptoms and an antibody response occur simultaneously, and other diagnostic tests are generally unavailable. Although serologic studies may be used retrospectively to diagnose infections due to HSV, VZV, or CMV, these results are seldom available in time to be useful in managing the acute infection or in guiding antiviral chemotherapy. Newer diagnostic techniques, such as detection of viral nucleic acids using radiolabeled probes, may become useful in the future for the rapid diagnosis of EBV or CMV infection.

Antibodies, Viral↗
PubMed1984

Pathologic changes of experimental simian varicella (Delta herpesvirus) infection in African green monkeys (Cercopithecus aethiops).

Gross and microscopic lesions of experimental simian varicella (Delta herpesvirus) infection in African green monkeys (Cercopithecus aethiops) are described. In stratified squamous epithelium, such as the epidermis, tongue, and esophagus, focal areas of epidermal hyperplasia, ballooning degeneration, and blood-filled vesicle formation with ulceration were apparent. Most visceral organs were involved, and the changes included necrosis, hemorrhage, and characteristic intranuclear eosinophilic inclusions in a variety of cells. A generalized vascular involvement was present with intranuclear inclusions in the endothelial and smooth muscle cells in various organs. The nervous system was normal on gross and microscopic examination. Virus titer and serum transferase values were correlated with the clinical signs of infection.

Animals↗
PubMed1977

Cell-mediated immune responses in recurrent herpesvirus infections. I. Lymphocyte proliferation assay.

Studies in immunosuppressed and immunodeficient patients indicate that the cell-mediated immune response appears to be responsible for controlling reactivated herpesvirus infections. In this study, the various parameters of a herpesvirus (types 1 and 2) antigen specific lymphocyte proliferation assay were optimized and used to evaluate individuals with clinical, recurrent HSV-1 and HSV-2 infections. Normal individuals with neutralizing antibody to HSV-1 or HSV-2 responded to virus antigen in culture as well as individuals with recurrent disease. Normal individuals without neutralizing antibody responded with a significantly lower response. Specificity of the lymphocyte proliferation assay was observed most strikingly in normal individuals with a rare HSV-1 infection during the vesicular eruption. Specificity was also observed by determining the ratio of the response to HSV-1 as compared to the response to HSV-2. Evaluated in this manner, individuals with recurrent HSV-1 infections had significantly higher ratios than individuals with HSV-2 infections and vice versa. Data from individuals with recurrent disease was compared to that of normal individuals to determine whether the former demonstrated a specific alteration in this response. Individuals with recurrent disease were found to have higher neutralizing antibody titers than normals. The neutralizing antibody titers in normal individuals correlated well with the lymphocyte proliferation assay results, whereas a similar evaluation in individuals with recurrent disease gave a negative correlation. The ratio of HSV-1 response/HSV-2 response also demonstrated a suppressed response in recurrent infections to the homologous virus during active disease, which disappeared when the individual was convalescent. These studies indicate that individuals with recurrent HSV infections have virus antigen specific alterations of their cell-mediated immune response, which can be associated with their disease.

Antibody Specificity↗
PubMed2005

Reactivation of koi herpesvirus infections in common carp Cyprinus carpio.

Two co-habitation studies with common carp were conducted to determine whether latent infections of koi herpesvirus (KHV) exist. Fish were exposed to KHV using 2 different temperature profiles, which induced low and high initial mortality. Subsequently, certain groups of fish were co-habited with naive fish while others were not. Koi herpesvirus was reactivated in fish from 3 of the 5 experimental tanks. Reactivation of the virus occurred regardless of the initial mortality associated with the virus or whether fish were co-habited with naive fish. The reactivation of the virus in our experiments occurred several months after the initial exposure to KHV and appeared to be temperature dependent.

Animals↗
PubMed1999

Comparison of PCR, virus isolation, and indirect fluorescent antibody staining in the detection of naturally occurring feline herpesvirus infections.

Cats with clinical signs suggestive of ocular infection with feline herpesvirus type 1 (FHV 1) and cats without such signs were assayed by 3 methods to detect FHV. Comparison of polymerase chain reaction (PCR), virus isolation, and indirect fluorescent antibody staining techniques for the detection of FHV demonstrated higher sensitivity of PCR in detecting this common infectious agent of cats. Compared with PCR, sensitivity and specificity for virus isolation was 49% and 100%, respectively, and those of indirect immunofluorescence were 29% and 96%, respectively. FHV was detected in 13.7% of client-owned cats with conjunctivitis and in 31% of shelter cats with no ocular signs. The use of FHV PCR as a diagnostic test for FHV-associated disease is limited because of the occurrence of healthy carriers.

Animals↗
PubMed1998

Nested polymerase chain reaction and in situ hybridization for diagnosis of canine herpesvirus infection in puppies.

The usefulness of two nucleic acid detection systems in suspected cases of spontaneous canine herpesvirus (CHV) infection in puppies was evaluated. Formalin-fixed, paraffin-embedded tissues from seven 1-3-week-old naturally infected puppies with lesions characteristic of CHV infection were investigated in a retrospective study. Nested polymerase chain reaction (PCR) and nonradioactive in situ hybridization (ISH) were used to detect nucleotide sequences of the CHV thymidine kinase (TK) gene. According to the original necropsy reports, CHV was isolated in four of the seven puppies using primary canine lung and/or kidney cells. In all seven puppies, gross and histologic lesions consisted of disseminated focal necroses and hemorrhages predominantly in kidneys, lung, liver, and spleen. In addition, few small amphophilic intranuclear inclusion bodies were detected by light microscopy mainly in epithelial cells of kidney, lung, and liver. ISH was performed with a 111-base-pair (bp) digoxigenin-labeled double-stranded DNA probe. Viral DNA was detected in the nuclei of cells near and within lesions. Various cell types, including bronchiolar and alveolar epithelial cells, hepatocytes, renal tubular epithelial cells, neurons, fibrocytes, cardiac myocytes, and endothelial cells, were positive for viral DNA. PCR amplification products of the expected length of 168 bp containing the expected cleavage site for the restriction enzyme EcoRI, derived from paraffin blocks containing lung, kidney, and liver tissues, were detected in all seven puppies. The specificity of the obtained amplicon was further confirmed by Southern blot analysis. ISH and PCR are both useful methods for diagnosing CHV infection in formalin-fixed, paraffin-embedded tissues and are highly specific and sensitive methods for further investigations of the pathogenesis of CHV-induced lesions.

Animals↗
PubMedDate not supplied

[Effect of experimental herpesvirus infection on morphofunctional state of mast cells].

Mast cells are involved in the reaction of the organism to herpesvirus infection, which manifests by changes in their count, morphology, and function. The onset of infection is characterized by primary activation of mast cells, the peak of disease by suppressed function, and convalescence by secondary activation and stabilization of cell morphology and function. Viral infection running in the presence of immunosuppression caused by cyclophosphamide is characterized by deep persistent suppression of the morphology and functions of mast cells at the peak of disease.

Animals↗
PubMed1980

Polymorphonuclear neutrophil-mediated antibody-dependent cell cytotoxicity of herpesvirus-infected cells: ultrastructural studies.

Bovine polymorphonuclear neutrophils (PMN) can mediate antibody-dependent cell cytotoxicity (ADCC) of herpesvirus-infected cells. Since cytotoxicity occurs only in the presence of PMN and specific antiviral antibody, but not until viral membrane antigens are expressed on the target cell, it is concluded that antibody must recognize viral membrane antigens before cytotoxicity can occur. Cytotoxicity also requires very close contact between the target cell and the PMN cell. These interactions occur as early as 1 h after incubating antibody, infected cells, and PMN, but the actual lysis and release of intracellular components occur over an extended period. It was assumed that degranulation was not involved in the initiation of cytotoxicity, but was involved in the final stage of destruction. The mechanism of lysis is proposed to involve the interaction of PMN membranes with target cell membranes with subsequent reorganization and activation of the PMN plasma membrane at points of contact with the target cell. This results in possible production of transmembrane channels which allows for the release of target cell contents.

Animals↗
PubMed1980

Complement-dependent, polymorphonuclear neutrophil-mediated cytotoxicity of herpesvirus-infected cells: possible mechanism(s) of cytotoxicity.

Highly enriched bovine polymorphonuclear neutrophils (PMN) were able to destroy herpesvirus-infected cells in the presence of complement. Our results suggest that some viral antigens are capable of activating the alternate complement pathway directly which results in cytotoxicity however, the addition of PMN plus complement resulted in high levels of cytotoxicity. Chelation of Ca2+ or Mg2+ indicated that Ca2+ was involved in PMN--target cell interactions but Mg2+ was involved in activation of the alternate complement pathway. Cytotoxicity in the presence of PMNs was shown to be under bidirectional control of cyclic nucleotides. Thus agents that elevated cAMP reduced cytotoxicity whereas agents that elevated GMP increased it. If the PMN is secreting some product during the cytotoxic reaction it must be performed since chemical shown to inhibit DNA, RNA and protein synthesis had no influence on CDNC. Our results are discussed in terms of the mechanism of CDNC lysis as well as in the vivo significance of such a defence mechanism.

Animals↗
PubMed1984

Diagnosis of herpesvirus infections: correlation of Tzanck preparations with viral isolation.

Evaluation of Giemsa-stained smears of scrapings from the base of vesicles or ulcers ( Tzanck preparation) for the presence of multinucleated giant cells and/or intracellular inclusions were diagnostic for herpesvirus in 18 of 21 cases (86%) of culturally proved herpesvirus infections. Smears from four patients with varicella-zoster infection also revealed cytologic alterations characteristic of herpesvirus.

Diagnostic Errors↗
PubMed2002

Improvement of serological discrimination between herpesvirus-infected animals and animals vaccinated with marker vaccines.

Control/eradication plans of bovine herpesvirus 1 (BHV1) and suid herpesvirus 1 (SHV1) infections involve vaccination with inactivated or attenuated gE-deleted marker vaccines and associated companion serological tests to discriminate naturally infected from vaccinated animals. Blocking or competitive enzyme-linked immunosorbent assays (ELISAs) have been designed for the detection of specific antibodies against BHV1 or SHV1 gE glycoprotein. The antigen source usually consists of a crude viral preparation in which gE is associated with other envelope glycoproteins. Such assays suffer from a lack of specificity which is not due to serological cross-reactions with other pathogens. Interestingly, false-positive results occur with sera collected from multivaccinated cattle or pigs. After multivaccination with a marker vaccine, the binding of the conjugated monoclonal antibody used as a tracer, could be hampered by antibodies directed against the other viral glycoproteins. In order to validate the steric hindrance hypothesis, a simple preadsorption of such samples was carried out with a preparation of antigen devoid of gE, prior to the blocking ELISA itself. The decrease in antibody concentrations against the major glycoproteins, clearly leads to a better discrimination between positive and negative samples; that is between infected and multivaccinated animals, without significant loss of sensitivity. This experiment confirms the steric hindrance hypothesis, therefore serum preadsorption could be an easy way to improve the specificity of currently available diagnostic tests.

Animals↗