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. 2011 Jun 28;6(6):e21674. doi: 10.1371/journal.pone.0021674

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This is an open-access article free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.

This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration, which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.

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PBMCs from seven M. bovis-BCG-vaccinated calves (black bars) and four non-vaccinated calves (grey bars) were treated with 10 µg/ml of M. bovis PPD and 0 or 100 ng/ml 25(OH)D₃ as indicated for 24 h. After treatment, PBMCs were sorted by FACS according to CD3/4/8/γδTCR (CD3; T cell), IgM (B cell), and CD14 (monocyte) expression on the cell surface. The amount of 1α-OHase (A), VDR (B), 24-OHase (C), iNOS (D), RANTES (E), IFN-γ (F), IL-17A (G), and IL-17F (H) mRNA in the sorted cells was determined by quantitative real-time RT-PCR. The amount of mRNA for each gene was normalized to the amount of RPS9 mRNA in each sample. Expression of each gene in each cell type is relative to non-stimulated, non-sorted PBMCs. Error bars represent SE. * P<0.05, *** P<0.001, Means are different.