A polyethylene catheter was inserted into the right carotid artery

p isolates was also previously observed in bovine macrophages. Similarly, a lack of correlation between 9 Interaction of Map Isolates with Ovine MDMs SU6668 web genotype and growth rate in the Bactec MGIT 960 system was observed for the ovine isolates; with C, S and B isolates from sheep growing at equivalent rates in MGIT cultures. Previously, it was suggested that a bovine, a bison, and a human type-C isolates induced anti-inflammatory and antiapoptotic responses in bovine MDMs, which would favour bacterial survival and persistence. In the current study, we showed that the successful survival of bovine and ovine isolates of Map within ovine MDMs correlated with an increased expression of BCL2-1, TIMP-1, TGFb-1, and IL10. In addition, a reduced proinflammatory immune response mediated by IL2 and TNFa-2 was observed in the infected cells. Our results also demonstrated that ovine macrophages infected with ovine isolates of distinct genotype did not differentially express BCL2-1, TIMP-1, TGFb-1, IL10, IL2 and TNFa-2. Previously, the bovine isolate-6 of Map that grew within BoMac cells was reported to induce the expression of the anti-inflammatory cytokines TGFb-1 and IL10 in BoMac cells that antagonized the proinflammatory response by down-regulating the production of TNF-a which favour bacterial survival. In regard to the apoptotic response, BoMac cells infected with the bovine isolate-6 had increased levels of expression of the apoptotic inhibitor BCL2-1 and of the inhibitor of tissue destruction TIMP-1 which might cause low levels of apoptosis. Overall, a strong correlation between the successful survival of Map isolates within bovine and ovine macrophages and the patterns of production of BCL2-1, TIMP-1, TGFb-1, TNFa-2 and IL10 was observed. Our results also indicated that the proinflammatory cytokines IL1a and IL2 were down-regulated in BoMac cells and in ovine MDMs infected with bovine isolates of Map, respectively. It is well known that the expression of these cytokines in the presence of intracellular bacteria is one of the first steps leading to activation of macrophages and effective bacteria killing. Consistently with our results, other authors also observed an up-regulation of TGFb and/or IL10 after the infection of bovine MDMs with live Map that down-regulated the production of TNFa. Similarly, rapid intracellular macrophage growth rates by strains of M. tuberculosis strains correlated with rapid production of IL10 that antagonizes the proinflammatory response by down-regulating the production of TNFa in THP-1 cells during the early stages of PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19660665 infection. Although MDMs can provide insights into Map-host interactions at the very early stages of the infection, this cellular model is unable to mimic later stages of the infection, such as early granuloma formation. To address this issue, three-dimensional in vitro models of granuloma have been recently developed. The formation of small, rounded granuloma-like structures was previously reported by coculture of human blood lymphocytes with autologous macrophages infected with live M. tuberculosis, M. leprae, or M. bovis or stimulation with mycobacterial antigens such as purified protein derivatives or lipomannan. In the present study, we reported for the first time the development of an in vitro model of ovine granuloma using Map-infected PBMCs cultured in an extracellular matrix composed of fibronectin and collagen, components of the surrounding tissue in which the natural granuloma is anchore