The facultative-pathogenic M. avium induced a profoundly different host cell signaling response P-gp inhibitor when compared to the non-pathogenic M. smegmatis [14]. In particular, the infection with M. smegmatis led to an increased p38 and ERK1/2 MAPKs activity in BMDMs which was Selleckchem GSK2118436 necessary for increased TNF secretion [14]. Furthermore, this increase in MAPKs was dependent upon prolonged stimulation of calmodulin/calmodulin kinase and cAMP/protein kinase A pathways [15]. In addition, sphingosine

kinase, phosphoinositide-specific phospholipase C and conventional protein kinase C were all implicated in M. smegmatis-induced activation of Erk1/2 [16]. One downstream target of the MAPK p38 was determined to be the transcription factor cyclic AMP response element binding protein (CREB) which was more activated in M. smegmatis-infected cells [17]. In order to understand why non-pathogenic mycobacteria are strongly attenuated we compared their capacity to induce ACP-196 an innate IR to that of facultative-pathogenic mycobacteria.

The induction of apoptosis and the stimulation of TNF expression in macrophages were analyzed and in both cases the macrophage response was much stronger for the non-pathogenic mycobacteria than the facultative-pathogenic mycobacteria. The induction of TNF secretion was important for the increase in caspase-3-dependent host cell apoptosis in BMDM. Furthermore, purified PI-LAM of the nonpathogenic mycobacterial species interacted with the TLR-2 and induced apoptosis and IL-12 p40 expression, whereas the purified Man-LAM of the facultative-pathogenic mycobacteria had no such activity. Altogether, facultative-pathogenic mycobacteria induce less of an innate

immune response in macrophages relative to non-pathogenic mycobacteria. Results and Discussion Non-pathogenic mycobacteria induce increased host cell apoptosis In order to test the Decitabine molecular weight apoptotic response of macrophages following infection with facultative-pathogenic compared to non-pathogenic mycobacteria, we used bone marrow-derived macrophages (BMDM) from BALB/c mice and infected them with M. smegmatis, M. fortuitum, M. bovis BCG, or M. kansasii for two hours. We then incubated the macrophages in infection medium with gentamycin for an additional twenty hours. The percentage of apoptotic cells was determined by quantifying the fraction of hypodiploid positive cells via flow cytometry (Figure 1A). 75-80% of BMDMs infected with M. smegmatis and M. fortuitum were hypodiploid positive which was significantly different (p < 0.001) from BMDMs infect with facultative-pathogenic mycobacteria (Figure 1B). Indeed, BMDMs infected with BCG and M. kansasii did not show any significantly increased levels of apoptosis compared to the untreated control cells during the course of this short term infection (p > 0.05; Figure 1B). Figure 1 Differences in apoptosis induced by facultative-pathogenic versus non-pathogenic mycobacteria in primary murine macrophages.