ABSTRACT
The incidence of herpes simplex virus type 1 (HSV-1) and herpes simplex virus type 2 (HSV-2) in herpes simplex encephalitis (HSE) was investigated using cerebrospinal fluid (CSF) samples from sixty-four cases of HSE. A polymerase chain reaction (PCR) employing primers flanking a region of the HSV thymidine kinase gene common to both HSV-1 and HSV-2 was used to detect HSV in the CSF. HSV-1 and HSV-2 were differentiated by digestion with restriction enzymes. Two enzymes were employed; Aval which cleaved only the HSV-2 gene product and Avall which cleaved only the HSV-1 gene product. Sixty-three cases of HSE were found to be due to HSV-1; one case due to HSV-2. These data confirm previous observations that HSV-2 is a rare cause of post-neonatal herpes encephalitis but indicates that a PCR procedure capable of detection of both viruses is essential for efficient diagnosis of HSE.
Subject(s)
Encephalitis, Viral/virology , Herpes Simplex/virology , Herpesvirus 1, Human/isolation & purification , Herpesvirus 2, Human/isolation & purification , Adolescent , Adult , Aged , Child , Child, Preschool , DNA, Viral/cerebrospinal fluid , DNA, Viral/genetics , Deoxyribonucleases, Type II Site-Specific , Encephalitis, Viral/diagnosis , Encephalitis, Viral/epidemiology , Female , Herpes Simplex/diagnosis , Herpes Simplex/epidemiology , Herpesvirus 1, Human/genetics , Herpesvirus 1, Human/pathogenicity , Herpesvirus 2, Human/genetics , Herpesvirus 2, Human/pathogenicity , Humans , Infant , Male , Middle Aged , Polymerase Chain Reaction , United Kingdom/epidemiologyABSTRACT
In this study we aimed to determine the incidence of herpes simplex virus (HSV) in the lungs of burns patients, and its association with the presence of adult respiratory distress syndrome (ARDS) and pneumonia. Haematoxylin and eosin (H&E), and immunohistochemical (IHC) staining for HSV was performed on lung tissue from 54 patients who had died following burn injury and from nine control cases. Polymerase chain reaction (PCR) for HSV deoxyribonucleic acid (DNA) was performed on a subset both of burns cases and controls. No viral inclusions were detected in H&E sections, but 50% of the burns cases were positive for HSV by IHC staining; no control cases were positive. Nuclear and cytoplasmic immunopositivity for HSV was seen in macrophages and epithelial lining cells. HSV was strongly associated with ARDS (p=0.007), but not with pneumonia (p=0.577). The relative risk of HSV infection was higher for cases with ARDS (2.21) than for those with pneumonia (1.26). PCR for HSV DNA was positive in three out of five burns cases, and in one out of five control cases. Immunohistochemical staining is more sensitive for the detection of herpes simplex virus than haematoxylin and eosin staining for detection of viral inclusions. Burns cases have a high incidence of pulmonary herpes simplex virus infection. Polymerase chain reaction results may not be fully representative due to problems of tissue necrosis postmortem. Pulmonary herpes simplex virus is strongly associated with adult respiratory distress syndrome and the two may be causally linked. Early detection and treatment of pulmonary herpes simplex virus in burns patients may reduce pulmonary complications and mortality.
Subject(s)
Burns/complications , Herpes Simplex/complications , Lung Diseases/complications , Adolescent , Adult , Aged , Aged, 80 and over , Burns/virology , Female , Humans , In Situ Hybridization , Lung/virology , Male , Middle Aged , Pneumonia/complications , Polymerase Chain Reaction , Respiratory Distress Syndrome/complications , Simplexvirus/isolation & purificationABSTRACT
AIMS/BACKGROUND: Herpes simplex virus (HSV) may establish latent infection in the cornea and therefore be transmissible by corneal transplantation. Monitoring of donor cornea culture medium was evaluated for HSV infection. METHODS: HSV was sought using virus isolation in cell culture, and its DNA was amplified to detectable levels using the polymerase chain reaction (PCR). RESULTS: Virus isolation in cell culture was negative on neat, cell pellet, and cell free supernatant prepared from the spent culture media of 80 corneas. Three cell pellets (3.8%) were positive for HSV DNA. The PCR positive culture negative results might have reflected latent rather than active HSV infection of the cornea. Post transplant follow up of the three recipients of corneas with HSV PCR positive organ culture media revealed no evidence of HSV induced eye disease or primary graft failure. CONCLUSION: Screening of corneal culture medium for HSV by virus culture or for HSV DNA by PCR could not be recommended.
Subject(s)
Culture Media/chemistry , DNA, Viral/isolation & purification , Simplexvirus/isolation & purification , Aged , Aged, 80 and over , Base Sequence , Corneal Transplantation , Female , Humans , Male , Middle Aged , Molecular Sequence Data , Polymerase Chain Reaction , Simplexvirus/genetics , Treatment Outcome , Virus LatencyABSTRACT
Five cases of apparent relapse of herpes encephalitis were investigated. All patients recovered after antiviral and corticosteroid therapy. Samples of CSF taken from the patients at intervals through the initial and subsequent encephalitic episode were examined. PCR amplification of a 351 bp sequence from the Herpesvirus simplex (HSV) thymidine kinase gene demonstrated the presence of HSV DNA in CSF taken during the initial encephalitic illness but not during the second encephalitic episode. Intrathecal synthesis of HSV antibody (HSV antibody index > 1.9) was observed in all cases following the first episode, and there appeared to be no significant increase in intrathecal antibody synthesis in the second episode. High levels of CSF myelin basic protein were found during the acute phases of both the initial and the subsequent encephalitic illnesses. These data suggest that at least in our series of five patients, relapse following HSE may not be due to active viral replication.
Subject(s)
Encephalitis, Viral/physiopathology , Herpes Simplex/physiopathology , Herpesvirus 1, Human/genetics , Myelin Basic Protein/cerebrospinal fluid , Adolescent , Adult , Aged , DNA, Viral/cerebrospinal fluid , Encephalitis, Viral/therapy , Encephalitis, Viral/virology , Female , Herpes Simplex/therapy , Herpes Simplex/virology , Humans , Male , Middle Aged , Polymerase Chain Reaction , RecurrenceABSTRACT
Three corneoscleral discs (from two donors) underwent subtotal endothelial loss during routine "long-term" organ culture storage. Laboratory studies of these corneas revealed evidence of herpes simplex virus (HSV) infection. The fellow cornea from one of the donors had been issued for transplant to a patient with keratoconus. Deterioration of the graft was noted 5 days after surgery; the disc was removed at 2 months and was shown to be infected with HSV. In an experiment designed to simulate initial "cleansing" of donor globes, 0.1% polyvinylpyrolidone-iodine protected cells from infection with HSV. It was concluded that the detection of HSV in these corneas could not be explained by external contamination of the ocular surface. Furthermore, culture of conjunctival and pharangeal swabs taken from 47 consecutive donors confirmed that HSV is rarely isolated at or around the time of death. Five pairs of donor corneas destined for use in transplantation were selected at random and investigated for the presence of HSV. HSV DNA was detected by polymerase chain reaction (PCR) in tissue from two of the corneal donors. Sequential stepwise sectioning suggested that HSV DNA when present was distributed in discrete foci within the cornea. These observations suggest that HSV infection may be a cause of severe endothelial loss during corneal organ culture and possibly provide an explanation for some "failures" of corneal grafting.
Subject(s)
Cornea/virology , DNA, Viral/analysis , Herpesvirus 1, Human/isolation & purification , Keratitis, Herpetic/transmission , Tissue Donors , Adult , Aged , Antigens, Viral/analysis , Cornea/pathology , Corneal Transplantation , Female , Graft Rejection/pathology , Graft Rejection/virology , Herpesvirus 1, Human/genetics , Herpesvirus 1, Human/immunology , Humans , Keratitis, Herpetic/pathology , Male , Middle Aged , Polymerase Chain ReactionABSTRACT
A number of techniques for extraction of DNA prior to polymerase chain reaction (PCR) amplification of herpes simplex virus (HSV) DNA in cerebrospinal fluid (CSF) were compared to the use of "native" CSF in the PCR reaction. The results indicate that extraction of DNA (which allows efficient removal of inhibitors of Taq polymerase) is an essential pre-requisite of the PCR detection of CSF HSV DNA.
Subject(s)
DNA, Viral/cerebrospinal fluid , Encephalitis/diagnosis , Herpes Simplex/diagnosis , Polymerase Chain Reaction/methods , Simplexvirus/genetics , Encephalitis/microbiology , Herpes Simplex/cerebrospinal fluid , HumansABSTRACT
T cell antigen specificity is determined by the products of the genes which encode the variable regions of their receptors. Of the T cell receptor (TCR) variable region gene products examined, only V beta 6.7a TCR-positive lymphocytes were reduced in primary Sjögren's syndrome patients with IgG1 hypergammaglobulinaemia compared with an age-, sex- and HLA-matched control population. The levels of V beta 6.7a T cells were also significantly reduced when these patients were compared with an age- and sex-matched but HLA-unmatched control group and non-tissue typed normal people of both sexes. Since published studies show no such abnormality in rheumatoid arthritis, systemic lupus erythematosus or other autoimmune diseases, this abnormality may reflect a pathogenic process specific to primary Sjögren's syndrome.
Subject(s)
Hypergammaglobulinemia/immunology , Immunoglobulin G , Sjogren's Syndrome/immunology , T-Lymphocytes/immunology , Adult , HLA Antigens/analysis , HLA-DR Antigens/analysis , Humans , Middle Aged , Phenotype , Receptors, Antigen, T-Cell/analysisABSTRACT
Thirteen patients with expansion of an unusual subset of T lymphocytes, defined by large size, cytoplasmic granularity and CD3+ CD8+ Leu 7+ surface phenotype, are reported. Although morphologically and/or phenotypically abnormal lymphocytes were found in all patients, only five had an absolute peripheral blood lymphocytosis. Ten patients had a bone marrow lymphocytosis. As in previous series, there was a strong association with neutropenia (12 patients) and polyarthropathy (seven patients). The latter group displayed a wide range of articular disease: classical or definite rheumatoid arthritis in four patients and milder non-erosive disease in the remainder. All 13 patients showed evidence of abnormal B cell function: IgM rheumatoid factor was present in nine patients, neutrophil-specific antibodies in six and all showed an increased level of at least one immunoglobulin isotype. These patients may be difficult to distinguish from those with idiopathic neutropenia and Felty's syndrome. Such a distinction may not be made on clinical grounds alone: critical assessment of lymphocyte morphology, bone marrow examination and analysis of lymphocyte phenotype should be considered in all patients with unexplained neutropenia, particularly in the context of arthritis. It is suggested that the true prevalence of this syndrome may have been greatly underestimated.
Subject(s)
Joint Diseases/diagnosis , Neutropenia/diagnosis , T-Lymphocyte Subsets/immunology , Aged , Aged, 80 and over , Antigens, Differentiation/analysis , Arthritis/diagnosis , Arthritis/immunology , CD3 Complex/analysis , CD8 Antigens/analysis , Female , Humans , Joint Diseases/immunology , Male , Middle Aged , Neutropenia/immunology , SyndromeABSTRACT
Herpes simplex virus thymidine kinase gene specific polymerase chain reaction (PCR) amplification of DNA extracted from lumbar cerebrospinal fluid (CSF) and Southern blotting (SB) were evaluated as a method for the diagnosis of herpes simplex encephalitis (HSE). Positive PCR-SB results were obtained with CSF samples from 9 of 10 patients (11 of 12 CSF specimens) with proven herpes encephalitis as early as 2 days after onset of neurological illness. Our data support the suggestion that PCR techniques may provide a clinically relevant "non-invasive" method for the diagnosis of HSE.
Subject(s)
DNA, Viral/cerebrospinal fluid , Encephalitis/diagnosis , Herpes Simplex/diagnosis , Simplexvirus/genetics , Base Sequence , Blotting, Southern , Encephalitis/complications , Herpes Simplex/complications , Humans , Molecular Sequence Data , Polymerase Chain Reaction , Simplexvirus/enzymology , Thymidine Kinase/geneticsABSTRACT
This research attempt to clarify and quantify the ways in which heroin addicts deal with interpersonal relations. A sample of heroin addicts on a methadone maintenance program were given the Fundamental Interpersonal Relations Orientation-Behavior (FIRO-B) scale. An accidental sample consisted of 35, approximately one-half, of the clinic's population. A mean interpersonal orientation profile is established and interpreted. An analysis of variance was performed for each of the six FIRO-B scores. Patients needs based on FIRO-B scores are discussed with regard to the descriptions of addicts life-styles as described in the literature. The possible etiological basis for these interpersoanl orientations is also discussed. Several therapeutic approaches, as indicated by FIRO-B scores, are explored.