TLR2 and TLR4 genes increased 6.54-fold and 5.28-fold, respectively, in the healthy control group. However, the expression of IL10 (2.90-fold) 3-h poststimulation was less compared than that seen in the stimulated tuberculosis group (8.74-fold). We compared the gene expression levels in the two groups. The results showed that two of the seven genes examined (TLR2 and IL10) were differentially

expressed in both the stimulated tuberculosis subjects and the stimulated healthy control subjects (P≤0.05 by t-test). Although TLR2 showed increased expression in Sotrastaurin both stimulated groups, it had a greater fold increase in the stimulated control group (6.54-fold) over the stimulated tuberculosis group (2.64-fold). This may indicate that TLR2 plays a larger role in regulation in healthy animals. In contrast, IL10 expression in stimulated tuberculosis animals (8.74-fold) was greater than that seen in the stimulated control group (2.90-fold). Thus, TLR2 may

play a key role in the response of MDMs from healthy cattle to M. bovis stimulation, while IL10 may play a similar key role during M. bovis stimulation of MDMs from tuberculosis cattle. The CPE and the relationship between M. bovis and MDMs cells were observed directly by microscopy (Fig. 2) and Ziehl–Neelsen stain (Fig. 3). The present findings learn more demonstrate that the CPE could be seen under microscopy after 3 h of stimulation, and it became

more severe over time. Necrosis and detachment Protein kinase N1 of cells were caused by the intrusion and adherence of bacteria. At 3 h, the medium incubated with cells was almost clear and intracellular fast-acid bacteria were seldom seen. However, by 10 h, the medium became unclear due to cellular debris and dead cell granules and bacteria could be observed inside the cytoplasm and the nucleus of some cells. Twenty-four hours after stimulation, the massive cellular death made microscopy images obscure and only a small number of cells survived. The M. bovis used in this study was a virulent strain, triggering a strong interaction and quickly leading to massive cellular death. Based on the observations made by microscopy and fast-acid stain, there are no obvious differences in CPE of M. bovis between MDMs from tuberculosis and healthy control cattle. The growth and survival status of intracellular M. bovis were assessed by bacterial CFU in the MDMs from tuberculosis and healthy control cattle (Fig. 4). Our data indicate that at 3 h, the CFU of intracellular survival of M. bovis is very low and difficult to measure in several subjects (less than three bacterial clones on a 7H10 agar plate). This result is consistent with the previous observation, by microscopy and fast-acid stain, that M. bovis grows poorly in cells after 3 h of infection. This study also shows that 10 h after stimulation, CFUs of M.