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  • 1
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary We constructed and expressed different overlapping fusion proteins with the nef gene of HIV-1 and generated specific polyclonal rabbit and monoclonal mouse antibodies against these recombinant proteins. The rabbit antisera, one of the monoclonal antibodies as well as a serum from a HIV-1 infected patient recognized the nef protein with Mr 27 kDa in latently HIV-1 infected glioma cells in the immunoblot. In contrast, these antibodies could not detect nef in productively HIV-1 infected Molt-3 cells neither in immunoblot nor in indirect immunofluorescence assays. These results indicate the possible participation of nef in viral latency. The recombinant nef proteins were used as probes for anti-nef antibodies in human sera. We observed in 17 of 57 sera tested specific anti-nef antibodies. All of these anti-nef positive sera also contained antibodies directed against viral structural proteins. The NH2-terminal region of the recombinant nef was shown to be the major immunodominant antigenic site in the immunoblot assay.
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  • 2
    ISSN: 1433-9285
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract   Background: We tested the hypothesis that the political change occurring in East Germany after the fall of the Berlin Wall in November 1989 affected patients with long-term schizophrenia, resulting in more and longer hospitalisations. Methods: In two samples in East Germany (120 patients in East Berlin, 70 patients in Chemnitz) and in a control group from West Berlin (40 patients), hospitalisations for each month between November 1984 and October 1994 were assessed. Each sample included all schizophrenia patients who were in continuous treatment in the given community mental health centre throughout the full observation period. Results: Hospitalisation indices were not higher after November 1989 than before in any of the groups. Conclusions: There is no evidence that political change in East Germany negatively affected the course of long-term schizophrenia as assessed by hospitalisations. If consistent medical care is provided, characteristics of the political system may have less impact on the course of schizophrenia than is sometimes assumed.
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  • 3
    ISSN: 1432-1440
    Keywords: AIDS (acquired immunodeficiency syndrome) ; Expression cloning ; HIV-1 (human immunodeficiency virus type 1) ; Immunofluorescence ; Recombinant protein ; Westernblot
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Serologic testing for human immunodeficiency virus type 1 (HIV-1) is currently based on enzyme linked immunosorbent assay (ELISA) as screening method. Positive ELISA-results have to be confirmed by at least one second procedure such as Western blotting or immunofluorescence. To obtain new diagnostic reagents for confirmatory testing, we expressed viral antigens in procaryotic systems. Peptides representing epitopes of structural core (gag)- and envelope (env)-proteins of HIV were produced inE. coli as stable immunogenic β-galactosidase fusion proteins. Recombinant proteins were taken for immunoblot-assays. The results of Western blotting with those fusion proteins were in general comparable with conventional ELISA, immunofluorescence, immunoblot with cell-culture derived virus and commercially available ELISA tests based on recombinant proteins. Immunoblots using recombinant transmembrane protein (gp41) derived polypeptide were more sensitive than the conventional procedure with purified virion proteins. Western blotting with recombinant fusionproteins provide reliable and inexpensive serodiagnostics without handling of infectious cell cultures.
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  • 4
    ISSN: 1432-1440
    Keywords: Cytomegalovirus ; Procaryotic expression ; Recombinant antigen ; Phosphorylated protein pp150 ; Monoclonal antibody ; Immunofluorescence ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Procedures for diagnostics of cytomegalovirus infections include histopathology, cell culture, serology, and direct detection of viral antigens or nucleic acids within infected cells or tissues. In order to develop a new diagnostic reagent for viral antigen detection, we generated a mouse monoclonal antibody. This antibody was raised against a recombinant antigen representing part of the large phosphorylated structural protein pp150 of human cytomegalovirus. The monoclonal antibody was shown to be useful for antigen detection by immunofluorescence and immunoenzymatic staining in infected cells from cell culture as well as from infected organs. The antibody proved to be reactive even in paraffin-embedded sections from tissue specimens.
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  • 5
    ISSN: 1432-1831
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Immunization studies with HSV-1 and HSV-2 envelope proteins expressed in Escherichia coli were performed. After active immunization of mice with a gD-1 antigen (Leu53-Ala312) expressed as a fusion protein, the animals were protected from a lethal challenge with HSV-1 and HSV-2. In addition, antisera from rabbits immunized with the same gD-1 antigen also conferred passive immunity to mice against a challenge infection with either HSV-1 or HSV-2. In contrast to these successful gD-1 protection experiments, various gC-1 and gC-2 fusion proteins from E. coli failed to induce protective immunity. Moreover, the mice sera from immunized animals were not able to react with the authentic, glycosylated gC-1 and gC-2 envelope proteins, whereas sera raised against authentic gC-1 and gC-2 glycoproteins do recognize the gC fusion proteins from E. coli. These results indicate, that E. coli might represent an ideal system for expressing gD antigens as a possible component of a HSV vaccine, whereas gC antigen cannot be produced in an immunocompetent form in E. coli.
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