Immunological study of the nef protein from HIV-1 by polyclonal and monoclonal antibodies.
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.
Kienzle
N
N
Abteilung Virologie, Institut für Medizinische Mikrobiologie und Hygiene, Homburg, Federal Republic of Germany.
Bröker
M
M
Harthus
H P
HP
Enders
M
M
Erfle
V
V
Buck
M
M
Müller-Lantzsch
N
N
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001483:Base Sequence; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D015693:Gene Products, nef / Q000096:biosynthesis / Q000276:immunology*; D015483:HIV Antibodies / Q000276:immunology*; D015658:HIV Infections / Q000276:immunology; D015497:HIV-1 / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009841:Oligonucleotides; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000276:immunology; D054311:nef Gene Products, Human Immunodeficiency Virus
D000911:Antibodies, Monoclonal; D015693:Gene Products, nef; D015483:HIV Antibodies; D009841:Oligonucleotides; D011993:Recombinant Fusion Proteins; D054311:nef Gene Products, Human Immunodeficiency Virus