Use of synthetic peptides for the detection of antibodies against the nef regulating protein in sera of HIV-infected patients.
Human sera were tested for the presence of anti-nef antibodies by radioimmunoassay (RIA), with recombinant radiolabelled nef expressed in E. coli. Of the 300 HIV-positive sera tested by RIA, 70 +/- 5.3% were found to be anti-nef positive. Anti-nef antibodies bound to nef with a high affinity (K 0.5 = 2.2 x 10(-9) M). In 31 of the sera, the specificity of anti-nef antibodies was further analysed by enzyme-linked immunosorbent assay (ELISA) with large synthetic peptides ranging from 31 to 66 amino acid residues and spanning the total sequence of nef from HIV-1. The results obtained showed that the immunodominant antigenic sites of nef were located close to the N- and C-terminal regions of the molecule.
Sabatier
J M
JM
CNRS UA 1179-INSERM U 172, Laboratoire de Biochimie, Faculté de Médecine Nord, Marseille, France.
Clerget-Raslain
B
B
Fontan
G
G
Fenouillet
E
E
Rochat
H
H
Granier
C
C
Gluckman
J C
JC
Van Rietschoten
J
J
Montagnier
L
L
Bahraoui
E
E
D000163:Acquired Immunodeficiency Syndrome / Q000276:immunology*; D000595:Amino Acid Sequence; D005260:Female; D019204:GTP-Binding Proteins / Q000235:genetics / Q000276:immunology; D015693:Gene Products, nef; D005809:Genes, Regulator; D006678:HIV / Q000235:genetics / Q000276:immunology* / Q000502:physiology; D015483:HIV Antibodies / Q000032:analysis*; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012191:Retroviridae Proteins / Q000235:genetics / Q000276:immunology*; D014779:Virus Replication; D054311:nef Gene Products, Human Immunodeficiency Virus
D015693:Gene Products, nef; D015483:HIV Antibodies; D010455:Peptides; D011994:Recombinant Proteins; D012191:Retroviridae Proteins; D054311:nef Gene Products, Human Immunodeficiency Virus; D019204:GTP-Binding Proteins