Mycobacterium bovis is an important pathogenic bacterial species and the causative agent of most cases of tuberculosis in cattle. Bovine tuberculosis (BTB) continues to pose a threat to livestock worldwide and also has serious implications

for human health (Tiruviluamala & Reichman, 2002). Macrophages are the major host cell of M. bovis infection and they mediate the host immune response to BTB through pathogen recognition and activation of an inflammatory response (Casadevall, 2008; Ahmad, 2011). Studies have shown differential gene expression between animals with tuberculosis and healthy animals (MacHugh et al., 2009; Moller NVP-BKM120 datasheet & Hoal, 2010; Maertzdorf et al., 2011). These studies have implicated innate immune responses, TLR signaling and Th1 cytokines in the cellular response to M. bovis (Werling & Jungi, 2003; Netea et al., 2005). Innate immunity relies heavily on the behavior

of inflammatory molecules. Proinflammatory cytokines TNF-α, IL1, and its receptor IL1R1, play an important role in the innate immune response, which is an essential mechanism for host defense against invading M. bovis (Salgame, 2005). TLR2 and TLR4 could recognize BTB products and rapidly generate a defensive response involving numerous proinflammatory cytokines and Th1 cytokines to restrict the growth of intracellular M. bovis. IL10 could downregulate the Th1 response and upregulate Th2 response in pathogen–host interaction, which may lead to the lack of protection of the host from M. selleck kinase inhibitor bovis (Jacobs et al.,

2000). We hypothesized that macrophages from tuberculosis infection and healthy cattle may have distinctive immune regulation and gene expression PAK6 in response to M. bovis stimulation. In this study, we use a monocyte-derived macrophages (MDMs) model to study the interaction of M. bovis and macrophages from tuberculosis cattle as well as healthy controls. Using real-time quantitative PCR, the expression of seven genes implicated in BTB (IL1β, IL1R1, IL1A, TNF-α, IL10, TLR2 and TLR4) was examined in MDMs from tuberculosis and healthy control cattle stimulated with M. bovis, respectively. We also observed the cytopathic effect (CPE) caused by M. bovis stimulation in MDMs cells by microscopy directly. Using Ziehl–Neelsen staining, bacteria entering into MDMs cells were detected to obtain a general impression of pathogen–host interaction. The growth and survival status of intracellular M. bovis may directly reflect the capability of cells in resisting and killing intracellular bacterium. We assessed the survival status of intracellular M. bovis by bacterial CFU in the MDMs to see the difference in the bacterial load between MDMs from tuberculosis and healthy control cattle. Virulent M. bovis Beijing strain (bovis strain 93006) was received from the China Institute of Veterinary Drug Control (CVCC, China). This is a wild-type strain isolated from tuberculosis lesions of a tuberculin skin test-positive cow in Beijing in 1953.