Mary Devlin

ISOLATION OF HERPES SIMPLEX VIRUS FROM HUMAN TRIGEMINAL GANGLIA, INCLUDING GANGLIA FROM ONE PATIENT WITH MULTIPLE SCLEROSIS

Herpes-simplex virus (H.S.V.) was isolated from 18 of 39 trigeminal ganglia (T.G.) obtained within 12 h of death. The virus was isolated from ten persons who had died of trauma, from one case of lymphoma, and from one case of multiple sclerosis. In the cadaver with histologically confirmed multiple sclerosis, large bilateral areas of demyelination were present near the points of entry of the nerve root, and the possibility that H.S.V. migration to the root entry zone caused demyelination cannot be excluded.

Extraction of cell-associated varicella-zoster virus DNA with Triton X-100-NaCl

Varicella-zoster virus (VZV) DNA was extracted from infected cells with 0.25% Triton X-100-0.2 M NaCl and purified by isopycnic centrifugation in CsCl. In each of eight experiments, 1.8-9.8 micrograms VZV DNA was obtained from 107 infected cells. The VZV DNA obtained by this procedure had a molecular weight of 88-100 x 106 as determined by sucrose gradient sedimentation and electron microscopy, and cleavage patterns after digestion with four restriction enzymes that corresponded to patterns previously described with six strains of VZV; the pattern of BamHI-cleaved Triton-NaCl-extracted VZV DNA was identical to the pattern seen after DNA extraction from virions. These studies expand the usefulness of Triton X-100-NaCl for extraction of large molecular weight viral DNA from a system where considerable cell-free virus is produced (Pignatti et al., 1979, Virology 93, 260) to a system known for its marked cell association.

Persistence of varicella-zoster virus DNA in blood mononuclear cells of patients with varicella or zoster

Varicella-zoster virus (VZV) DNA was detectable by in-situ hybridization in blood mononuclear cells (MNCs) of patients with varicella or zoster for 2–56 days after the onset of a rash. VZV DNA was present in many MNCs from one acute varicella patient 2 days after the onset of the rash and was rarely found in MNCs during acute zoster, convalescent zoster, and convalescent varicella. The morphology of MNCs containing VZV was heterogenous, although most viral-DNA-containing MNCs were large monocytoid cells. Serial examination of blood MNCs from one adult with varicella revealed VZV DNA up until 8 weeks, but not 16 weeks, after the appearance of the rash; parallel studies in four zoster patients showed VZV DNA up until 3 weeks, but not later than 7 weeks after the appearance of the rash. These results indicate that MNCs become infected with VZV during the primary encounter with VZV (varicella) and during reactivation (zoster) and that infection continues for weeks after the onset of the skin rash. Furthermore, the detection of VZV DNA in blood MNCs of uncomplicated zoster patients coincides with the period during which these patients experience pain.

The production of varicella zoster virus antiserum in laboratory animals

SummaryHyperimmune anti-varicella zoster virus (VZV) antisera were prepared in BALB/c mice, Lewis rats and New Zealand rabbits by subcutaneous inoculations of purified VZ virions suspended in Freunds adjuvant. VZV antibodies were demonstrated both by an enzyme-linked immunosorbent assay (ELISA) and by indirect immunofluorescence (IF). VZV could not be detected in organs of any inoculated animals by IF or by cocultivation of tissue fragments from infected animals with human diploid fibroblasts.

Affinity-purified varicella-zoster virus glycoprotein gp1/gp3 stimulates the production of neutralizing antibody

Varicella-zoster virus glycoprotein gp1/gp3 was purified by affinity chromatography using anti-gp1/gp3 monoclonal antibody 19.1 linked to CNBr-activated Sepharose CL-4B. Rabbits immunized with purified glycoprotein gp1/gp3 developed mono-specific neutralizing antibody.

The internal organization of the varicella-zoster virus genome.

DNA was extracted from varicella-zoster (VZ) virions prepared in sucrose gradients. Thirty-eight molecules examined by electron microscopy were found to have a mean length of 46.7 micrometers. Examination of self-annealed VZV DNA molecules revealed that the virus genome was composed of a unique linear large sequence with a mol. wt. of 74.4 X 10(6) to 78.4 X 10(6), and a unique short sequence of mol. wt. approx. 9.8 X 10(6) flanked by inverted repeat sequences of 4.7 X 10(6) mol. wt.

Human brain in tissue culture. VI.Presence of glial fibrillary acidic protein in subcultivated human fetal brain cells as demonstrated by immunofluorescent and immunoperoxidase staining

SummaryGlial fibrillary acidic protein (GFAP) was present in cell cultures derived from human fetal brain tissue as determined by indirect immunofluorescence (IF) and immunoperoxidase (IP) staining using rabbit anti-human GFAP antisera. The IF and IP techniques were comparable in localizing the cytoplasmic distribution and the frequent perinuclear concentration of GFAP in brain cells. The horseradish peroxidase technique was more sensitive and a 1:20–1:40 dilution of anti-GFAP serum could be applied in the initial step of the peroxidase staining as compared to a 1:10 dilution of anti-GFAP serum for IF staining. Sequential studies of subcultivated human fetal brain cell lines by these techniques indicated that some brain cell lines become GFAP-negative rapidly, whereas other cell lines remain GFAP-posifive for no less than ten subcultivations in vitro. GFA protein was never present in any PMLSV40-transformed human brain cells.

Varicella-Zoster Virus Latency in The Nervous System

The cause of multiple sclerosis (MS) is not known. The main hypotheses are that the disease is produced by a virus or is a virustriggered immunopathology. Although evidence to support the theory that MS is caused by a virus is only presumptive, there are four compelling reasons to search for virus in MS tissue. First, although MS does not occur until adult life, epidemiologic studies suggest that the causative agent is acquired in childhood.[1] An exposure period before or at the time of puberty followed by a long latent period before the onset of the disease is known to occur with other types of infection that lead to clinical CNS disease.[2] Furthermore, a point-source epidemic of MS occurred on the Faroe Islands from 1943-1960 during the British occupation;[3] thus MS on the Faroes would appear to be transmissible and probably infectious. Secondly, the cerebrospinal fluid (CSF) of patients with MS contains oligoclonal bands and increased levels of immunoglobulin (Ig)G irrespective of the amount of total protein. Only a few other diseases in humans are associated with oligoclonal bands and increased CSF-IgG; many are inflammatory and have also been shown to be infectious.[4] Third, progressive multifocal leucoencephalopathy is a human demyelinating disease in which a papovavirus produces a lytic infection of oligodendrocytes.[5] Fourth, numerous DNA and RNA viruses have been shown to be capable of producing demyelinating disease in rodents after experimental inoculation.[6]