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. 2014 Sep 10;8(2):414–428. doi: 10.1038/mi.2014.79

Figure 7.

Figure 7

CD83 homotypic interaction mediates anti-inflammatory responses through p38 mitogen-activated protein kinase. (a) siRNA knockdown of CD83 abrogates response to CD83.Fc or αCD83 antibody. Enzyme-linked immunosorbent assay (ELISA) of interleukin (IL)-12p40 shows CD83 required on surface of cells to mediate response. Double asterisks (**) indicate CD83.Fc is significantly different from mature dendritic cell (mDC) control, P=0.0085. Triple asterisks (***) indicate αCD83 is significantly different from mDC control, P<0.0001. (b) Truncated CD83 lentiviral expression constructs. (c) ELISA of IL-12p40 production. Lentiviral overexpression of full-length (FL) human CD83 does not inhibit MUTZ3 DC response to CD83; however, expression of the cytoplasmic truncated CD83 blocks inhibitory effect of CD83. Fc protein and αCD83 antibody. Triple asterisks (***) indicate CD83.Fc is significantly different from mDC control, P<0.0001. Double asterisks (**) indicate αCD83 is significantly different from mDC control, P=0.0037. (d) CD83 chimeric lentiviral expression constructs. Extracellular domain of CD79a fused to CD83 transmembrane region (TM) and FL or truncated cytoplasmic domain. (e) ELISA of IL-12p40 production. Lentiviral overexpression of FL chimera inhibits MUTZ3 DC response to anti-CD79a antibody, while expression of truncated chimera abrogates this response. Triple asterisks (***) indicate anti-79a treated significantly different from control mDC, P<0.0001. Cross-linking alone is sufficient to drive anti-inflammatory response through CD83 cytoplasmic domain. (a, c, e) Results representative of three independent experiments. (f) Western blots of monocyte-derived dendritic cell (MDDC) lysates show p38 and signal transducer and activator of transcription factor 3 (STAT3) phosphorylation upon stimulation and CD83 treatments for 5 min. Phosphorylation of p38 is decreased in cells treated with CD83.Fc or αCD83 antibody. No differences seen in phosphorylation of STAT3. Total p38 and total STAT3 used as a loading controls. Results representative of MDDCs generated from three independent donors.