CD4 changes conformation upon ligand binding.
Aurintricarboxylic acid (ATA) has been shown to block the binding site for both HIV gp120 and mAb anti-Leu 3a on CD4. We have unexpectedly found that brief treatment with > or = 1 micrograms/ml ATA rapidly disengages another mAb, OKT4E, after it has been bound to CD4 on human PBL. OKT4E is specific for a discontinuous epitope overlapping the MHC class II-binding region in the N-terminal CD4 domain. Interestingly, among 10 other mAb tested, only anti-Leu 8, specific for a leukocyte homing receptor is also quickly released from the cells by ATA treatment. Disengagement of the OKT4E mAb is also seen on a CD4-positive cell line (HPB-ALL) and with recombinant soluble CD4 (sCD4) bound to immobilized OKT4E. In all of these cases, disengagement is prevented if OKT4E is cross-linked, or the Leu 3a site is blocked by the mAb, but not by gp120. Photobleaching fluorescence resonance energy transfer (pFRET) measurements suggest that OKT4E is released as an indirect consequence of ATA-evoked conformational changes of CD4. Similar changes were detected as a result of gp120 binding to PBL. These data raise the possibility of a novel type of immunomodulation: induced disengagement of a bound ligand from its Ag.
Szabò
G
G
Center for Drug Evaluation and Research, FDA, Washington, DC 20204.
Pine
P S
PS
Weaver
J L
JL
Rao
P E
PE
Aszalos
A
A
D000911:Antibodies, Monoclonal / Q000276:immunology; D000937:Antigen-Antibody Reactions / Q000187:drug effects*; D001312:Aurintricarboxylic Acid / Q000494:pharmacology*; D015704:CD4 Antigens / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D008024:Ligands; D008560:Membrane Fluidity; D008856:Microscopy, Fluorescence; D011487:Protein Conformation
D000911:Antibodies, Monoclonal; D015704:CD4 Antigens; D008024:Ligands; D001312:Aurintricarboxylic Acid