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  • 1
    ISSN: 1432-0533
    Keywords: Neoplasms ; Cerebellar diseases ; Cerebellum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Sera from patients with paraneoplastic cerebellar degeneration have been shown to contain high titers of antibody to human Purkinje cells. It is not known, however, whether these sera react with cerebellar material from species other than man. The present study was conducted to determine whether cerebellar tissue of nonhuman species could be used to screen human sera for anticerebellar antibodies and whether similarities in cerebellar antigens between nonhuman and human material might permit attempts to transmit cerebellar degeneration to experimental animals by passive transfer of patient sera. Sections of human, monkey, pig, sheep, cat, rabbit, and rat cerebellar tissue were overlaid with serial dilutions of positive sera from patients with cancer and with paraneoplastic cerebellar degeneration and were stained using indirect immunofluorescence methods. All sera stained specific Purkinje cells when reacted with monkey, pig, and rabbit cerebellar sections, but not all sera stained sheep, cat, or rat tissue. Immunofluorescent labeling of animal cerebellar tissue was less bright than that obtained with human cerebellar sections, and anticerebellar antibody titers were invariably lower when assayed with nonhuman than with human material. Although human anticerebellar antibodies react with cerebellar tissue from other animal species, patient-to-patient variation in staining is sufficiently great that not all patient sera might be suitable for passive transfer experiments, and that attempts to identify anticerebellar antibodies by reacting patient sera with nonhuman cerebellar tissue could be negative where these antibodies are in fact present and could be demonstrated using human material.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0533
    Keywords: Papovaviruses ; JC virus ; Progressive multifocal leukoencephalopathy ; T antigen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Formalin-fixed, paraffin-embedded autopsy sections of brains from two patients with progressive multifocal leukoencephalopathy (PML) were stained by peroxidase-antiperoxidase methods for human papovavirus T antigen, a nonstructural protein expressed in cells lytically infected or transformed by JC, BK, and SV40 viruses. Adjacent sections were stained for papovavirus common structural antigen, a component of JC, BK, and SV40 virions which is synthesized in productively infected but not transformed cells. Intense immunoperoxidase labelling specific for T antigen was detected in large numbers of oligodendrocytes at the edges of demyelinated areas and in occasional oligodendrocytes within otherwise normal brain. Occasional morphologically normal astrocytic cells exhibited similar specific staining, but only rate atypical astrocytic cells contained detectable amounts of T antigen. Examination of adjacent sections stained with antisera to common structural antigen revealed an identical pattern of immunoenzymatic labelling, indicating that most of the cells expressing T antigen were also expressing viral structural proteins. The present study demonstrates that T antigen can be identified by immunoperoxidase methods in routinely processed autopsy material from cases of PML, but that detectable amounts of antigen are found almost exclusively in cells undergoing lytic infection.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Archives of virology 94 (1987), S. 169-173 
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Murine K-papovavirus was serially passaged 10 times in primary cultures of mouse embryo cells, using cell suspensions and media from infected cultures to inoculate fresh flasks at each passage level. Titers of K virus infectivity in cell suspensions and media rose during serial passage, but viral hemagglutinating activity was not detected in cells or media before the 10th passage. Although the infectivity of K virus for mouse embryo cultures was enhanced by serial passage, lethality of the virus for suckling mice was lost. The present study represents the first successful serial transmission of K virus infectionin vitro.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Primary cultures of lungs, kidneys, and glial cells derived from mid-gestation Syrian hamsters were inoculated with 105 hemagglutinating units of murine K-papovavirus and were serially subcultivated to allow appearance of lines of persistently infected or transformed cells. K virus did not replicate in renal cell cultures and produced only transient productive infection of lung cells. Evidence of K virus-induced cell transformation was not detected in either of these cultures. Inoculation of glial cultures with K virus, however, resulted initially in a protracted infection in which 80–100 percent of cells expressed K virus V antigen for 18 subcultivations and in which cloning experiments suggested that all cells in the culture contained the viral genome. After 18 subcultivations numbers of positive cells rapidly dimished, and cells appeared which exhibited altered morphology and density dependence. These altered cells (KVHG3 cells) grew well in serum-free media, could be cloned in soft agar, and were negative for infectious virus or K virus V antigen. Although KVHG3 cells did not exhibit staining when reacted with antisera to K virus T antigen, Southern blot analysis of these cultures demonstrated the presence of K virus DNA integrated into the host chromosomal DNA and indicated that some rearrangement of the viral genome had occurred. Attempts to produce tumors in hamsters with these cells were unsuccessful, as were attempts to induce tumors in newborn hamsters by intracranial inoculation of K virus. The present study demonstrates that K virus is capable of causing productive infection and cell transformation in primary cultures of fetal hamster glial cells but that other hamster cell types are relatively resistant to the virus and that both K virus and K virus-transformed hamster cells are poorly oncogenic for hamstersin vivo.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-0533
    Keywords: Key words Purkinje cells ; Blood-brain barrier ; Human anticerebellar antibody ; Rat ; Paraneoplastic syndromes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Paraneoplastic cerebellar degeneration accompanying gynecological or breast malignancies is frequently associated with an autoantibody response, termed "type I" or "anti-Yo" directed against cytoplasmic antigens of cerebellar Purkinje cells. The role of this antibody response in the pathogenesis of paraneoplastic cerebellar degeneration is unknown; however, it is also not known whether anti-Purkinje cell antibodies from the systemic circulation bind to targe t Purkinje cell antigens under the conditions of brain inflammation and blood-brain barrier disruption, which are frequently present at the onset of cerebellar symptoms. Inbred Lewis rats received intraperitoneal injections of type I or normal IgG in the setting of blood-brain barrier disruption induced by adoptive transfer of experimental allergic encephalomyelitis (EAE) and were killed after 24, 48, and 96 h. Brains of these animals were studied histologically for evidence of EAE and immunohistochemically for binding of human or endogenous rat IgG to target neurons. Rat IgG was detected around vessels and in Purkinje cells of all animals studied. Human IgG was detected around vessels of all animals. In animals examined 96 h after receiving type I human IgG, human IgG was identified within processes of Purkinje cells and within occasional Purkinje cell bodies. Uptake of type I IgG by other cell types was not observed, and neuronal uptake of IgG was not seen in brains of animals receiving normal human IgG. Ou r data demonstrate that circulating type I IgG is internalized by cerebellar Purkinje cells in the setting of blood-brain barrier disruption and suggest a mechanism by which an antibody response directed against cytoplasmic antigens of Purkinje cells may reach target antigens at the onset of paraneoplastic cerebellar degeneration.
    Type of Medium: Electronic Resource
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