Soluble CD16/FcγRIII Induces Maturation of Dendritic Cells and Production of Several Cytokines Including IL-12
FcγRIII (CD16), a low affinity FcR which binds IgG-containing immune-complexes, exists under membrane-associated forms and under a soluble form (sFcγRIII). The latter, present in biological fluids (serum, saliva), is generated by proteolytic cleavage of the two membrane-associated FcγRIII isoforms, FcγRIII-A (expressed by macrophages and NK cells) and FcγRIII-B (expressed exclusively by neutrophils). Herein we demonstrate that dendritic cells (DCs), generated by culturing monocytes with GM-CSF and IL-4, bind biotinylated recombinant sFcγRIII. This binding is specific and involves the complement receptor CR3 (CD11b/CD18) and CR4 (CD11c/CD18). Indeed, preincubation of DCs with anti-CD11b and anti-CD11c mAbs decreased by 52% and 62% respectively the binding with sFcγRIII. Moreover, electron microscopy showed that binding of gold-labeled sFcγRIII to DCs maintained at 4°C occured within clathrin-coated pits. Once internalized, at 37°C, sFcγRIII entered the endocytic pathway and reached the MHC class II compartments. Furthemore, DCs incubated for 48 h with multivalent sFcγRIII expressed increased levels of CD40, CD80, CD86, CD54, CD58, HLA class I and class II molecules and decreased levels of CD23 and CD32. These effects result in an increased capacity of DCs to trigger proliferative responses by CD4+ CD45RA+ allogeneic T cells. RT-PCR amplification demonstrated that incubation of DCs for 20 h in the presence of multivalent sFcγRIII induced the appearance of GM-CSF and IL-12 p40 mRNA. Among the cytokines constitutively expressed, IL-lβ and IL-8 were strongly up-regulated whereas IL-6 and IL-12 p35 mRNA were increased to a lesser extent and the expression of MIP-lα mRNA remained constant. Finally, ELISA tests demonstrated that DCs incubated with multivalent sFcγRIII secreted the cytokines IL-1β, IL-6, IL-8, GM-CSF and IL-12 p75. Thus, while becoming internalized sFcγRIII could affect the capacity of DCs to present antigens and, via the induction of accessory molecules and the release of the IL-12 p75 protein, could initiate Thl type immune response.
KeywordsEndocytic Pathway Type Immune Response Induce Maturation Counterflow Centrifugation Elutriation Recombinant sCD16
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