Immunological study of the nef protein from HIV-1 by polyclonal and monoclonal antibodies |
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Authors: | N. Kienzle M. Bröker H. -P. Harthus M. Enders V. Erfle M. Buck N. Müller-Lantzsch |
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Affiliation: | (1) Abteilung VirologieAbteilung VirologieAbteilung Virologie, Institut für Medizinische Mikrobiologie und Hygiene, Homburg, Federal Republic of Germany;(2) Behringwerke AG, Marburg;(3) GSF-Abteilung für Molekulare Zellpathologie, Neuherberg, Federal Republic of Germany |
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Abstract: | 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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