Zofia Wroblewska

Retroviral sequences related to human T-lymphotropic virus type II in patients with chronic fatigue immune dysfunction syndrome.

Chronic fatigue immune dysfunction syndrome (CFIDS) is a recently recognized illness characterized by debilitating fatigue as well as immunological and neurological abnormalities [Straus, S.E. (1988) J. Inf. Dis. 157, 405-412]. Once thought to be caused by Epstein-Barr virus, it is now thought to have a different but unknown etiology. We evaluated 30 adult and pediatric CFIDS patients from six eastern states for the presence of human T-lymphotropic virus (HTLV) types I and II by Western immunoblotting, polymearse chain reaction, and in situ hybridization of blood samples. The majority of patients were positive for HTLV antibodies by Western blotting and for HTLV-II gag sequences by polymerase chain reaction and in situ hybridization. Twenty nonexposure healthy controls were negative in all assays. These data support an association between an HTLV-II-like virus and CFIDS.

Experimental demyelination produced by the A59 strain of mouse hepatitis virus

Intracerebral inoculation of 4- to 6-week-old C57BL/6 mice with the A59 strain of mouse hepatitis virus (MHV), a murine coronavirus, produced biphasic disease. Acute hepatitis and mild meningoencephalitis were followed by subacute spastic paralysis with demyelinating lesions in the brain and spinal cord as determined by Eponembedded toluidine-blue-stained sections and by electronmicroscopy. MHV-A59 was cultured by plaque assay from the blood, brain, spinal cord, and liver of infected mice during the acute phase, but not in the chronic stage. MHV-A59 antigen was detected by immunofluorescence (IF) until 3 months postinfection (PI). Serum anti-MHV-A59 antibodies were detected from 7 days to 5 months PI. The induction of demyelination by MHV-A59 provides a suitable system to study virus-induced demyelination further.

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.

Human brain in tissue culture: Part. 5. Identification of glial cells by immunofluorescence

Glial fibrillary acidic (GFA) protein was demonstrated by immunofluorescence in both primary and long-term cultures of adult human brain cells explanted from normal, neoplastic and non-neoplastic CNS diseases, and in mouse brain cells subcultivated in vitro. GFA protein was present in the cytoplasm of at least 4 distinct cell types and persisted throughout the finite lifetime of brain cell cultures.

CSF antibodies to myelin basic protein and oligodendrocytes in multiple sclerosis and other neurological diseases

ABSTRACT – Cerebrospinal fluid (CSF) from 18 multiple sclerosis (MS) patients, 13 subacute sclerosing panencephalitis (SSPE) patients, 22 other neurological disease (OND) patients, and 7 neurotic patients as controls were tested in an 125I‐labeled anti‐human F(ab′)2 binding assay for the presence of antibodies to normal human brain cells from tissue culture, human fibroblasts, plasma membranes of MS and normal human brain, myelin basic protein (MBP) and bovine oligodendrocytes. Antibodies to MBP and to oligodendrocytes were found in the CSF of MS, SSPE and OND patients. Absorption of CSF with bovine CNS myelin significantly diminished binding activity to oligodendrocytes. Antibodies in the CSF against MBP and oligodendrocytes, on which some myelin determinants are expressed, seem to be a common feature of diseases in which demyelination is a component.

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.

Varicella-zoster virus infection of human brain cells and ganglion cells in tissue culture

SummaryThe growth of varicella-zoster virus (VZV) in cultures of human brain (HB) and human ganglion (HG) cells was compared to VZV growth in human fibroblasts. Infected cultures were monitored by histologic, electron microscopic (EM), and virologic techniques. Two to three days after VZV infection of all cell cultures at a multiplicity of infection (MOI) of 0.1, a multifocal cytopathic effect (CPE) developed. CPE was characterized by multinucleated cells and virus-specific intranuclear inclusions as determined by immunofluorescence and EM. In VZV-infected HB and HG cells only, large vacuoles were also seen in the cytoplasm of dying cells. Some vacuoles were almost devoid of structures. Within and at the limiting membranes of other vacuoles, aggregates of VZV particles (measuring 210–230 nm) were seen enveloped in osmiophilic material. VZV infection of HB and HG cultures was strongly cell-associated. Clarified tissue culture medium removed at maximum CPE failed to infect homologous HB or HG cells. When an inoculum of VZV-infected HB or HG cells was transferred to homologous uninfected cultures for 10–15 passages, the incubation period for CPE remained constant, and the titer of VZV in cells sampled randomly corresponded to the amount of virus that was used for original infection.

Growth of JC virus in adult human brain cell cultures.

SummaryAdult human brain (AHB) cells infected with JC virus (JCV) developed a cytopathic effect (CPE) beginning 12–14 days after infection. Ultrastructurally, 37–40 nm papova virions were seen in the nuclei of infected cells, and both T and V antigen were demonstrated by indirect immunofluorescence. The hemagglutinating titer of JCV in infected AHB cells was 10–40 times higher than the amount of JCV used to initiate infection. AHB cells are more readily available than primary human fetal brain cells, they can be subcultured 15–25 timesin vitro and they support JCV replication after multiple subcultivations. These properties make the AHB cell line useful for propagating JCV.

Persistent parainfluenza type 1 (6/94) infection of brain cells in tissue culture

SummaryA state of persistent infection with parainfluenza type 1 virus (6/94 strain) was established in cultures of human and bovine brain cells. Following primary infection of human brain cells, viral cytopathic effect (CPE) and hemadsorption (HAD) depended on the multiplicity of infection. After persistent infection was established the virus rapidly became cell-associated; no CPE occurred and no viral antigen was detectable by HAD, immunofluorescence (FA), or immunoprecipitation. Infectious virus could be recovered only by fusion or cocultivation. This was in marked contrast with infected bovine brain cells, where, following primary infection, little or no CPE occurred. A productive infection rapidly evolved and persisted without CPE, but with 100 per cent HAD and FA positive cells.

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.