FcR gamma-chain is essential for both surface expression and function of human Fc gamma RI (CD64) in vivo.
Most Ig receptors exist as hetero-oligomeric complexes with separate ligand binding (alpha) and signal transducing (beta, gamma, or zeta) subunits. For Fc gamma RIIIa and Fc epsilon RI, association with the FcR gamma-chain is essential for surface expression. However, the human high affinity IgG receptor, hFc gamma RI, was found to be surface-expressed by itself in transient transfection models. We have now analyzed the integrity of hFc gamma RI expression in more detail in stable transfectants. In vitro we noted that, in the absence of FcR gamma-chain, surface expression of hFc gamma RI rapidly declined to background levels, in both IIA1.6 B cells and NIH3T3 fibroblasts. The effect of FcR gamma-chain on hFc gamma RI surface expression in vivo was evaluated by using two newly generated transgenic mouse lines, selectively expressing hFc gamma RI on myeloid cells. These transgenic mice were crossed with FcR gamma-chain-deficient mice. Analysis of blood monocytes and peritoneal macrophages showed that surface expression of hFc gamma RI was reduced by approximately 80%. The remaining approximately 20% of receptors were still capable of binding IgG-opsonized RBC, suggesting FcR gamma-chain not to be critical for hFc gamma RI ligand-binding capacity. Importantly, however, hFc gamma RI signaling capacity was lost in FcR gamma-chain-deficient cells. No phagocytosis could be observed using either ligand sensitized (EA-IgG2a) or CD64-targeted erythrocytes (using a bispecific antibody) in both hFc gamma RI transgenic lines. This documents the FcR gamma-chain to be indispensable for both surface membrane expression and function of human Fc gamma RI in vivo.
van Vugt
M J
MJ
Department of Immunology, University Hospital Utrecht, The Netherlands.
Heijnen
I A
IA
Capel
P J
PJ
Park
S Y
SY
Ra
C
C
Saito
T
T
Verbeek
J S
JS
van de Winkel
J G
JG
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D006801:Humans; D007142:Immunoglobulin gamma-Chains / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D017452:Receptors, IgG / Q000235:genetics / Q000276:immunology*; D015398:Signal Transduction
D007142:Immunoglobulin gamma-Chains; D017452:Receptors, IgG