Production and characterization of a human monoclonal antibody, reactive with a conserved epitope on gp41 of human immunodeficiency virus type I.
A human Epstein-Barr virus-transformed lymphoblastoid B-cell line was generated from peripheral blood mononuclear cells (PBMC) of an asymptomatic human immunodeficiency virus type I (HIV-1) seropositive donor, which produces a human monoclonal antibody K14 (IgG1), reactive with an epitope on the transmembrane part (gp41) of the envelope glycoprotein of HIV-1. This monoclonal antibody reacts with a lysate of HIV-1-infected H9 cells, gradient purified HIV-1, and a vaccinia recombinant HIV-1 gp160 protein, but not with HIV-2 antigens in an enzyme-linked immunosorbent assay (ELISA). When used as an immobilized ligand in an immune affinity column, K14 selectively purifies gp41 from a HIV-1-infected H9 cell lysate. Although no reactivity was observed in ELISA with a panel of partially overlapping synthetic nonapeptides spanning the whole length of HIV-1 gp41, it was shown to react with recombinant envelope proteins, provided that they did contain amino acids 643-692: deletion of this part resulted in the disappearance of the reactivity. Testing of an extensive panel of the sera from HIV-1 seropositive or seronegative donors from Europe and Africa, including a selected group of donors before and after HIV-1 seroconversion, in a competition ELISA with horseradish peroxidase-conjugated K14, showed that the epitope recognized on gp41 is immunodominant and conserved. K14 does not neutralize HIV-1 infectivity or virus-mediated cell fusion, and does not mediate antibody-dependent cellular cytotoxicity.
National Institute of Public Health and Environmental Protection, Laboratory of Immunobiology, Bilthoven, The Netherlands.
V J
VJ
Teeuwsen
K H
KH
Siebelink
S
S
Crush-Stanton
B
B
Swerdlow
J J
JJ
Schalken
J
J
Goudsmit
R
R
van de Akker
M J
MJ
Stukart
F G
FG
Uytdehaag
A D
AD
Osterhaus
D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology; D001667:Binding, Competitive; D002472:Cell Transformation, Viral; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis*; D015483:HIV Antibodies / Q000096:biosynthesis* / Q000276:immunology; D015497:HIV-1 / Q000276:immunology*; D004854:Herpesvirus 4, Human; D006801:Humans; D014759:Viral Envelope Proteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D015483:HIV Antibodies; D014759:Viral Envelope Proteins