fermentans and M. penetrans prevent apoptosis and stimulate host cell growth of infected cells whereas the predominantly surface-colonizing species M. hominis and M. salivarium promote apoptosis [33]. Inhibition of P2X7-signaling appears to be more important for intracellular pathogens as shown by the treatment of M. tuberculosis infected macrophages with ATP, which results in killing of both the intracellular mycobacteria and the host, whereas

conditions such as complement-mediated cytolysis, Fas ligation, and CD69 activation induced only lysis of the macrophages while preserving the bacterial vitality https://www.selleckchem.com/products/cilengitide-emd-121974-nsc-707544.html [34–36]. With regard to the findings that M. hominis, a well known colonizer of epithelial surfaces, has also been found in the intracellular compartment in cultured HeLa cells [37], Trichomonas vaginalis [38] and human spermatozoa [39], OppA-mediated cytoadhesion of M. hominis may play a key role in invasion. In case of infection the extracellular

ATP-level is increased. Thus, an OppA-mediated decrease of this danger signal, thus preventing P2X7 – mediated signaling, with concomitant cytoadhesion are proposed mechanisms for mycoplasma survival to circumvent host immune defense mechanisms and facilitate invasion. Conclusions The present study demonstrates that the enzymatic function of OppA as main ecto-ATPase of M. hominis is essential for adhesion and suggests that the unique feature of this mycoplasma has an impact on patho-physiological important processes in host-pathogen interactions. Methods CH5424802 datasheet Etomidate HeLa cell culture The human cervical carcinoma cell line HeLa S3 (ATCC CCL2.2) was obtained from the American Type Culture Collection (Rockville, MD, USA) and cultivated in Dulbecco’s Modified Eagle Medium (Invitrogen GmbH, Darmstadt, Germany) with 10% horse serum (PAA laboratories GmbH, Pasching, Austria.) Mycoplasma culture conditions and purification

of proteins The M. hominis strains FBG was grown in PPLO broth base medium containing 1% (w/w) arginine as described previously [40]. Stocks were prepared from a mid-logarithmic-phase broth culture and stored in 1 ml portions at -70°C. For the Selleckchem Ilomastat purification of distinct proteins, cells of 1 L mid-logarithmic-phase broth culture were sedimented (10.000 × g, 20 min, 4°C) and the sediment washed twice with PBS and resuspended in 10 ml PBS. After protein concentration was estimated by Bradford analysis [41] and adjusted to 1 mg protein/ml PBS, membrane proteins were solubilised by 0.5% (w/v) N-dodecylmaltoside (Roche, Grenzach- Wyhlen, Germany). After 1 h incubation on a rotation wheel followed by centrifugation (15.000 × g, 20 min, RT), the supernatant was incubated with sepharose-coupled antibodies DC10, BG2 or CG4 and the respective proteins OppA, P50 and P60/P80 were isolated as previously described [6]. Dephoshorylation of wild type OppA 2 μg OppA were incubated with 5 units shrimp alkaline phosphatase in 50 μl [10 mM Tris/HCl, pH 7.