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Background  Bovine tuberculosis (BTB) caused by  Mycobacterium bovis  continues to cause substantial losses to global agriculture and has significant repercussions for human health. The advent of high throughput genomics has facilitated large scale gene expression analyses that present a novel opportunity for revealing the molecular mechanisms underlying mycobacterial infection. Using this approach, we have previously shown that innate immune genes in peripheral blood mononuclear cells (PBMC) from BTB-infected animals are repressed  in vivo  in the absence of exogenous antigen stimulation. In the present study, we hypothesized that the PBMC from BTB-infected cattle would display a distinct gene expression program resulting from exposure to  M. bovis . A functional genomics approach was used to examine the immune response of BTB-infected ( n  = 6) and healthy control ( n  = 6) cattle to stimulation with bovine tuberculin (purified protein derivative – PPD-b)  in vitro . PBMC were harvested before, and at 3 h and 12 h post  in vitro  stimulation with bovine tuberculin. Gene expression changes were catalogued within each group using a reference hybridization design and a targeted immunospecific cDNA microarray platform (BOTL-5) with 4,800 spot features representing 1,391 genes.    Results  250 gene spot features were significantly differentially expressed in BTB-infected animals at 3 h post-stimulation contrasting with only 88 gene spot features in the non-infected control animals ( P  ≤ 0.05). At 12 h post-stimulation, 56 and 80 gene spot features were differentially expressed in both groups respectively. The results provided evidence of a proinflammatory gene expression profile in PBMC from BTB-infected animals in response to antigen stimulation. Furthermore, a common panel of eighteen genes, including transcription factors were significantly expressed in opposite directions in both groups. Real-time quantitative reverse transcription PCR (qRT-PCR) demonstrated that many innate immune genes, including components of the TLR pathway and cytokines were differentially expressed in BTB-infected ( n  = 8) versus control animals ( n  = 8) after stimulation with bovine tuberculin.    Conclusion  The PBMC from BTB-infected animals exhibit different transcriptional profiles compared with PBMC from healthy control animals in response to  M. bovis  antigen stimulation, providing evidence of a novel gene expression program due to  M. bovis  exposure.

Antigen stimulation of peripheral blood mononuclear cells from Mycobacterium bovis infected cattle yields evidence for a novel gene expression program