CD16/32-specific biotinylated 2.4G2 single-chain Fv complexed with avidin-FITC enhances FITC-specific humoral immune response in vivo in a CD16-dependent manner.
Fcgamma receptors (FcgammaRs) play an essential role in the regulation of immune response due to their ability to bind immune complexes. Activating FcgammaRs may facilitate antigen presentation and dendritic-cell maturation, while in the late phase of the immune response, the inhibitory FcgammaRIIb may down-regulate B-cell activation upon cross-linking with activating receptors. In this study, we investigated the in vivo role of FcgammaRs on the modulation of humoral immune response. In order to get well-defined immune complexes that can bind to both the activating and the inhibitory FcgammaRs, we designed a mono-biotinylated single-chain fragment variable construct from the rat anti-mouse CD16/32 clone 2.4G2, linked to avidin-FITC, and tested its effect on the FITC-hapten-specific T-independent type 2 (TI-2) and T-dependent (TD) immune response. When injected intravenously in mice, the complex bound to a small portion of B220+, CD11b(high) and CD11c(high) cells and was localized in the spleen on marginal zone macrophages 15 min after treatment. When applied as a booster following primary immunization with TI-2 (FITC-dextran) or TD (FITC-keyhole limpet haemocyanin) antigens, the complex elevated the number of hapten-specific IgM/IgG-producing B cells. This effect was diminished in CD16KO mice, suggesting that the activating-type FcgammaRIII might be a key mediator of this mechanism.
Angyal
Adrienn
A
Department of Immunology, Eotvos Lorand University, 1117 Budapest, Hungary.
Szekeres
Zsuzsanna
Z
Balogh
Péter
P
Neer
Zsuzsa
Z
Szarka
Eszter
E
Virag
Viktor
V
Medgyesi
David
D
Prechl
Jozsef
J
Sarmay
Gabriella
G
D000818:Animals; D000921:Antibody-Producing Cells / Q000187:drug effects / Q000276:immunology*; D001360:Avidin / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology; D019921:Biotinylation; D016207:Cytokines / Q000378:metabolism; D003911:Dextrans / Q000008:administration & dosage / Q000276:immunology; D016650:Fluorescein-5-isothiocyanate / Q000008:administration & dosage / Q000031:analogs & derivatives*; D006433:Hemocyanins / Q000008:administration & dosage / Q000276:immunology; D006825:Hybridomas; D056724:Immunity, Humoral / Q000187:drug effects; D007275:Injections, Intravenous; D008264:Macrophages / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D011485:Protein Binding; D051381:Rats; D017452:Receptors, IgG / Q000172:deficiency / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000276:immunology; D057127:Single-Chain Antibodies / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D013154:Spleen / Q000276:immunology; D013997:Time Factors
D016207:Cytokines; D003911:Dextrans; C499172:Fcgr2b protein, mouse; C485700:Fcgr3 protein, mouse; D017452:Receptors, IgG; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies; C015219:fluorescein isothiocyanate dextran; C036963:fluorescein isothiocyante avidin; D001360:Avidin; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin; D016650:Fluorescein-5-isothiocyanate