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. 2009 Jul 6;206(7):1603–1614. doi: 10.1084/jem.20090547

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© 2009 Cao et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jem.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 5. — BST2 activates primary pDCs and inhibits IFN and cytokine production by pDCs. (A) pDCs were incubated with anti-ILT7, recombinant Fc, or BST2-Fc proteins and then analyzed for calcium influx. pDCs pretreated with 5 µM of Syk inhibitor were also analyzed. Data are representative of three independent experiments. (B) The amounts of secreted cytokines from pDCs cultured with plate-bound Fc or BST2-Fc are shown. pDCs were activated overnight with either 0.2 µM of CpG 2216 or MOI 6 of Flu. Data are representative of five independent experiments with ten donors. (C) The levels of gene transcripts from pDCs cultured with purified Fc or BST2-Fc are shown. The relative expression of each gene was normalized with S18 and calculated against the value obtained from normal total PBMCs. Data are representative of two independent experiments. (D) BST2 does not affect co-stimulatory molecule expression by pDCs. pDCs cultured with plate-bound Fc or BST2-Fc and then activated with 0.2 µM of CpG 2216 for 48 h. Surface levels of CD80 and CD86 are shown. Data are representative of two independent experiments. (E) pDCs preincubated with medium, 20 µg/ml of IgG1 or 20 µg/ml of neutralizing ILT7 mAb were cultured overnight with plate-bound Fc or BST2-Fc in the presence of CpG 2216. The amounts of secreted IFN-α were measured by ELISA. Percent of BST2-mediated IFN-α suppression was calculated as percent ratio of IFN-α ([Fc minus BST2-Fc]/[Fc]) and plotted. Data are representative of two independent experiments with four donors. (F) The amounts of secreted IFN-α from Flu-challenged pDCs cultured with HEK293 with or without surface HA-tagged BST2 are shown. Data are representative of two independent experiments.