Characterization of an FcgammaRI-binding peptide selected by phage display.
The high-affinity IgG receptor, Fcgamma receptor I (FcgammaRI), is expressed exclusively on myeloid cells, and there is a great interest in the targeting of vaccine antigens to FcgammaRI using anti-human FcgammaRI antibodies or fragments derived from such molecules. In order to reduce the size and complexity of the targeting reagent, we have searched for FcgammaRI binding peptides in peptide libraries displayed on phage. The human monocytic cell line U937 was used as target. Phages that displayed the consensus peptide CLRSGXGC were selected and revealed increased binding to IFN-gamma stimulated versus non-stimulated U937 cells as well as to FcgammaRI transfected versus non-transfected IIA1.6 cells. Furthermore, they bound the extracellular domains of soluble FcgammaRI, but neither FcgammaRIIA, FcgammaRIIB nor FcgammaRIIIB. Binding was inhibited by a synthetic version of the peptide, whereas neither human IgG nor the FcgammaRI-specific monoclonal antibodies (mAb) mAb22 and 32.2 interfered. Flow-cytometry analysis and internalization studies showed that a synthetic biotin-conjugated peptide ADGACLRSGRGCGAAK-bio was able to target U937 cells and FcgammaRI transfected IIA1.6 cells, and further to promote internalization and vesicular degradation of streptavidin coupled to 1 microm magnetic beads. These peptides may have potential as FcgammaRI targeting reagents.
Berntzen
G
G
Department of Molecular Biosciences, University of Oslo, Norway.
Brekke
O H
OH
Mousavi
S A
SA
Andersen
J T
JT
Michaelsen
T E
TE
Berg
T
T
Sandlie
I
I
Lauvrak
V
V
D000911:Antibodies, Monoclonal / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D019084:Fluorescent Antibody Technique, Indirect; D005456:Fluorescent Dyes; D006801:Humans; D007074:Immunoglobulin G / Q000494:pharmacology; D007371:Interferon-gamma / Q000276:immunology; D019151:Peptide Library; D010455:Peptides / Q000302:isolation & purification / Q000378:metabolism*; D017452:Receptors, IgG / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D020298:U937 Cells
D000911:Antibodies, Monoclonal; C489017:FCGR1A protein, human; D005456:Fluorescent Dyes; D007074:Immunoglobulin G; D019151:Peptide Library; D010455:Peptides; D017452:Receptors, IgG; D007371:Interferon-gamma; D016650:Fluorescein-5-isothiocyanate