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Biomedical subjects

Z Wroblewska

Publications and source records attributed to Z Wroblewska.

At least 37 records · Page 2Linked to original sources

The production of varicella Zoster virus antiserum in laboratory animals. Brief report.

Hyperimmune 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 Freund's 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.

Animals↗

Demonstration of JC virus by immunofluorescence in multiple cell types in experimentally infected adult human brain cell cultures.

JC virus (JCV)-induced T antigen developed in the nuclei of approximately 50% of adult human brain (AHB) cells infected with JCV. Approximately 10% of these T antigen-positive cells contained glial fibrillary acidic protein (GFAP) and were identified as astrocytes in immunofluorescence studies. About 50% of these T antigen-positive cells bound antiserum to fibronectin on their surface and were identified as fibroblasts. No cells binding antiserum to galactocerebroside (GC), an oliodendrocyte-specific marker, were detected in infected or non-infected AHB cultures. the cell type of AHB that constitutes the remaining 40-50% of T antigen-positive cells that are GFAP-, fibronectin- and GC-negative remains to be determined.

Adult↗

Comparison of human cytomegalovirus growth in MRC-5 human fibroblasts, brain, and choroid plexus cells in vitro.

Cell cultures derived from human brain, choroid plexus, and human lung fibroblasts (MRC-5) were infected with the Towne strain of human cytomegalovirus (CMV). The cytopathic effect, beginning 24-48 hours after infection, was characterized by foci of enlarged rounded cells that spread slowly and eventually coalesced to destroy the entire monolayer within one week. Cowdry type A inclusion bodies and herpes virus nucleocapsids were seen in infected cells. CMV-specific antigen was demonstrated by immunofluorescence in fibroblasts and astrocytic cells of brain cultures and in cells of choroid plexus cultures as well as in MRC-5 fibroblasts. Despite these morphologic and immunochemical similarities the growth of CMV differed in cells of brain and choroid plexus origin as compared with MRC-5 cells. In brain and choroid plexus cell cultures most of the virus remained cell-associated throughout the observation period of one week, whereas in MRC-5 cells the CMV was found in both cell-associated and cell-free fractions of harvested material.

Antigens, Viral↗

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

Glial 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-positive for no less than ten subcultivations in vitro. GFA protein was never present in any PML-SV40-transformed human brain cells.

Animals↗

The use of vesicular stomatitis (visna virus) pseudotypes to demonstrate visna virus receptors in cells from different species.

Superinfection of visna virus (VV)-infected cells with vesicular stomatitis virus (VSV) resulted in the formation of a pseudotype virus population containing a VSV genome within a VV coat [designated VSV (VV)] as determined by plaque reduction neutralization with antisera to VSV and VV. These VSV (VV) virions were capable of infecting cell cultures from a number of species that were non-permissive for VV alone. Limited propagation of VV in some mammalian species would thus appear to be due to an intracellular restriction rather than to absence of VV receptors.

Animals↗

Herpes simplex type 1 DNA in human brain tissue.

Herpes simplex virus type 1 (HSV-1) is known to reside latently in the trigeminal ganglia of man. Reactivation of this virus causes skin lesions and may occasionally infect other tissues, including the brain. To determine whether the brain tissue of humans free of clinical signs of HSV-1 infection contains any trace of HSV-1, we examined the DNA from brain tissue by endonuclease digestion, separation of the fragments by gel electrophoresis, and hybridization with labeled HSV-1 DNA probes. Hybrid bands were detected autoradiographically in experiments using cloned and virion-purified fragments of the HSV-1 genome. HSV-1 DNA sequences were found in 6 of 11 human brain DNA samples tested. In some cases, these bands corresponded to the bands expected for the complete viral genome, whereas others contained bands representing only a part of the genome. In some cases, the terminal fragments could be found, suggesting that the DNA was in a linear, nonintegrated form.

Brain↗

Analysis of a predominant immunoglobulin population in the cerebrospinal fluid of a multiple sclerosis patient by means of an anti-idiotypic hybridoma antibody.

We have produced hybridoma antibodies directed against immunoglobulins present in the cerebrospinal fluid of a patient with multiple sclerosis (MS). One hybridoma antibody recognized an idiotypic determinant of an immunoglobulin population [an idiotype (Id)] which constituted approximately 1% of the immunoglobulin present in the cerebrospinal fluid. The Id focused in the pH range 8.0-8.4. It was present at roughly 10- to 15-fold higher relative (compared to total immunoglobulin) concentration in cerebrospinal fluid than in serum of the homologous MS patient. The Id could not be detected in three cerebrospinal fluid samples and 28 serum samples of heterologous MS patients or in the serum of 43 optic neuritis patients. The Id persisted in the homologous MS patient at increased concentration over the entire (6 years) observation period. The Id could be shown to react with a Theiler murine encephalomyelitis virus strain WW which was isolated from mice after inoculation with periplaque white matter from brain in a histologically confirmed case of MS.

Animals↗

Growth of JC virus in adult human brain cell cultures.

Adult 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 times in vitro and they support JCV replication after multiple subcultivations. These properties make the AHB cell line useful for propagating JCV.

Adult↗

Growth of murine cytomegalovirus in murine and heterologous brain cell cultures. Brief report.

Murine cytomegalovirus (MCMV) produced a cytopathic effect in mouse brain, guinea pig embryonic brain, human brain and fibroblast cells. Virus-specific antigen was detected by immunofluorescence in these cells after primary infection with MCMV. MCMV also replicated in ouse embryo brain and guinea pig brain cells. Although definite evidence of MCMV replication could not be demonstrated in human cells, MCMV infectivity was maintained for 12 days in human cell cultures.

Animals↗

Growth of the WW strain of Theiler virus in mouse central nervous system organotypic culture.

Myelinated organotypic cultures of mouse central nervous system (CNS) provide a favorable milieu for growth of the WW strain of Theiler virus (TV). WW-TV induced a cytopathic effect characterized by neuronal destruction and swelling of myelin sheaths. Tissue culture medium and homogenates of infected cultures produced neurologic disease in mice, and TV was demonstrated in cultures by indirect immunofluorescence and electron microscopy. Ultrastructurally, severe alterations in neurons and the presence of cytoplasmic inclusions containing paracrystalline arrays of 30-nm virus particles were evident in astrocytes. Demyelination appeared to be secondary to lysis of oligodendrocytes by virus. CNS organotypic cultures provide an environment for the study of TV-induced demyelination.

Animals↗

Acute demyelinating disease in a chimpanzee three years after inoculation of brain cells from a patient with MS.

Brain cells from a patient with classic multiple sclerosis were inoculated intracerebrally into the frontal lobe of a newborn chimpanzee. The animal developed acute quadriplegia three years, two months later and was killed four days after the onset of symptoms. Central nervous system lesions were primarily localized in the spinal cord at root entry zones; these were characterized by demyelination and regeneration of myelin by Schwann cells.

Animals↗

Detection by complementation of defective or uninducible (herpes simplex type 1) virus genomes latent in human ganglia.

Reconstruction experiments have shown that temperature-sensitive (ts) mutants of herpes simplex virus type 1 (HSV-1)(Glasgow strain 17) grow, complement, and recombine with similar efficiency in human nerve ganglion cells, human brain cells, normal human fibroblasts (WI38), and baby hamster kidney (BHK) 21/C13 hamster cells. Cultures of human trigeminal, superior cervical, and vagus ganglia that had failed to release herpes simplex virus spontaneously were superinfected with a range of ts mutants of HSV-1 and incubated at both permissive (31 degrees C) and nonpermissive (38.5 degrees C) temperatures. Progeny virus was assayed at both temperatures to determine if complementation of or recombination with the input genomes had occurred. The results showed that the ganglia from 8 of 14 individuals, which had been consistently negative for spontaneous release of virus, contained information that could be detected or rescued following superinfection with ts mutants of herpes simplex virus. In two additional cases, positive results were obtained after the superinfection of negative ganglia explants, but in each of these herpes simplex virus had previously been spontaneously released from one of six ganglia explanted.

DNA, Viral↗

Cytomegalovirus isolation from a chimpanzee with acute demyelinating disease after inoculation of multiple sclerosis brain cells.

A strain of cytomegalovirus (CMV) was isolated during the third subcultivation of explants from the left frontal lobe of a chimpanzee that developed paralysis more than 3 years after intracerebral inoculation at birth with brain cell cultures derived from a patient with multiple sclerosis. Another strain of CMV was also isolated from a lymph node culture taken from the same chimp. The isolates, designated MZM-13 and MZM-14, produced a cytopathic effect characteristic for CMV when inoculated into brain, ganglion, or fibroblast cultures of human or simian origin. Infected cells contained characteristic Cowdry A intranuclear as well as intracytoplasmic inclusion bodies, and 100-nm spherical herpes-like virus particles were detected by electron microscopy in the nucleus and cytoplasm of infected cells. Virus was further identified as CMV with convalescent human anti-CMV serum. Complement-fixing antibody to CMV was present at a titer of 1:32 when the acutely ill chimpanzee was sacrificed. No antibody was detected at birth or at 1 or 2 years of age. A newborn chimpanzee inoculated intracerebrally with MZM-13 developed clinically asymptomatic lesions in the central nervous system characterized by acute and chronic inflammation and degeneration of myelin in cranial and spinal nerve roots. Restriction endonuclease analysis of viral deoxyribonucleic acid isolated from these two viruses indicated that MZM-13 and MZM-14 are identical and are closely related to chimpanzee CMV. No similarity in restriction endonuclease fragment patterns was found between MZM virus and the Towne and Clegg strains of human CMV.

Animals↗

A technique for the elution of cell-surface antibody from human brain tissue.

An artificial system is described in which anti-dinitrophenyl (DNP) antibody attaches to hapten-conjugated human brain tissue. Treatment of this material with acetic acid at pH 2.5 to 3.0 for 90 seconds followed by immediate neutralization results in dissociation of antibody from the hapten-conjugated brain. Less than 1 ng of specific antibody can be detected, as determined by the ability of eluted material to combine with DNP in an established radioimmunoassay system. Similar methods may be applied to neurological disorders in which immunoglobulins are thought to be produced in the central nervous system.

Antibodies↗

Enhanced parainfluenza I (6/94) virus detection in latently infected human brain cell cultures by treatment with cytochalasin D and dimethyl sulfoxide.

The ability of cytochalasin D (CD) and dimethyl sulfoxide (DMSO) to enhance parainfluenza I (6/94) virus replication was studied in various cell culture systems. Treatment of CV1 cells with CD (1 microgram/ml) dissolved in DMSO prior to primary 6/94 virus exposure at 10(0)--10(5) multiplicities of infection did not substantially enhance virus replication. However, there was a transient increase in cell associated virus one day after infection of DMSO-treated cultures. CD treatment of cultures of human brain cells latently infected with 6/94 virus (LIHB cells) did not enhance 6/94 virus detection. Cocultivation of CV1 cells with CD-treated LIHB cell cultures, and cocultivation of LIHB cell cultures with CD-treated CV1 cells, resulted in the production of both cell-associated and cell-free 6/94 virus three and five days after cocultivation. No virus was detected after similar cocultivation of untreated LIHB cell cultures with untreated CV1 cells. The usefulness of CD-DMSO treatment in the rescue of virus from 6/94 LIHB cell cultures appears limited to a cocultivation system. The use of these techniques to enhance virus rescue from human tissues suspected of harboring latent viral genomes is discussed.

Animals↗

Rat tracheal organ culture supports replication of parainfluenza 1 (6/94) virus and promotes 6/94 virus rescue from latently infected human brain cells.

Rat tracheal organ culture (TOC) supported replication of parainfluenza 1 (6/94) virus. Cell-associated and cell-free viruses were found after primary infection of TOC. In contrast to other mammalian systems, rat TOC was capable of maintaining 6/94 virus infectivity after primary infection. Rat TOC may be considered a potential indicator system that could be used to detect virus latent in human tissue.

Animals↗