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Ligation of Dendritic Cell-Associated lectin-1 Induces Partial Maturation of Human Monocyte Derived Dendritic Cells

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Ligation of Dendritic Cell-Associated lectin-1 Induces Partial Maturation of Human Monocyte Derived Dendritic Cells

Elizabeth J Ryan et al. Hum Immunol.

Abstract

Dendritic cell-associated lectin-1 (DCAL-1), also known as C-type lectin-like-1 (CLECL1), is a novel C-type lectin-like molecule expressed by antigen presenting cells including dendritic cells (DCs). Here we report that incubation of immature DCs (iDCs) with an anti-DCAL-1 monoclonal antibody (mAb) induced downstream signaling, including phosphorylation of c-Jun N-terminal kinase (JNK) and p44/42 MAP kinase. Furthermore, ligation of DCAL-1 expressed by iDCs specifically enhanced HLA-DR expression, whereas the expression of other co-stimulatory molecules remained unchanged and minimal cytokine secretion was detected. DCs that express high levels of major histocompatibility complex (MHC) class II in the absence of high levels of other co-stimulatory molecules and inflammatory cytokine secretion may play an important role in the maintenance of immune tolerance. Therefore, our data suggests an important role for DCAL-1 in the regulation of the immune response.

Figures

Fig. 1
Fig. 1
Ligation of dendritic cell-associated lectin-1 (DCAL-1) induced the phosphorylation of downstream signaling molecules in dendritic cells (DCs). Immature DCs were incubated with an isotype matched control (IgM), anti-DCAL-1 (UW50), anti-CD40 (G28-5) or PMA/Ionomycin for the time points indicated, cell lysates were prepared, and the levels of phosphorylated proteins were determined by western blotting. (A) Stimulation of immature DCs with anti-DCAL-1 induces tyrosine phosphorylation. (B) Levels of specific phospho-proteins were analyzed. Total p-38 MAPK levels were used as a control for equal protein loading. The results shown are representative of two experiments performed with iDCs obtained from different donors.
Fig. 2
Fig. 2
Anti-dendritic cell-associated lectin-1 (anti-DCAL-1) treatment of immature dendritic cells (DCs) specifically upregulates HLA-DR. Immature DCs were treated with either 10 μg/ml of soluble Anti-DCAL-1, isotype control (mouse IgM) or 1 μg/ml of E. coli lipopolysaccharide as a positive control for 48 hours and the expression of DC maturation markers analyzed by flow cytometry. (A) Dot plots show the expression of CD1a versus CD83. (B) HLA-DR expression; the grey histograms represent the expression of HLA-DR following the different treatments and the black histogram indicates the staining of an isotype control antibody. Numbers represent the M.F.I. Isotype control versus anti-DCAL-1 treatment; p < 0.01 by Wilcoxon signed rank test. (C) CD86 (Numbers represent the M.F.I.), (D) CCR7 (Numbers indicate the percentage of cells expressing CCR7), (E) CCR5 (Numbers indicate the % of cells expressing CCR5). This experiment was performed on different donors (n = 10) with similar results and one of these experiments is shown.

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