asteroides growth and filament formation [23]. In human neutrophils, α-defensins HNP 1-3 are stored as active peptides in primary (azurophil) granules in concentrations of >10 mg/ml [24]. As granule-contents are minimally diluted after fusion with the phagocytic vacuole, HNP 1-3 targets ingested pathogens in concentrations multitudes higher than those needed for potent antinocardial killing observed in our study (LD90 of N. farcinca, N. nova and N. asteroides = 64 μg/ml). In

contrast, the human cathelicidin EPZ5676 ic50 is stored as inactive precursor hCAP-18 in secondary (specific) granules and is processed to LL-37 by proteinase 3 after secretion into the extracellular milieu. Like the human β-defensin hBD-3, LL-37 Saracatinib is additionally produced upon infection or inflammation by epithelial cells of the respiratory/gastrointestinal tract or by keratinocytes. Levels of LL-37 e.g. in airway surface fluids are estimated to be 1-5 μg/ml [25]. Concentrations of β-defensins are estimated to be in the range of 1-10 μg/ml [13]. Thus, in vivo concentrations of LL-37 and hBD-3 will most likely be not sufficient to exert direct nocardial killing. Nevertheless, LL-37 and hBD-3 may take part in antinocardial defense by additive or synergistic action with other antimicrobial peptides and proteins abundantly selleck products present along epithelial barriers. In favour of this hypothesis, we found additive killing of N. farcinica

in a model assay using a combination of LL-37 and HNP 1-3. Moreover,

owing to a wide range of biological activities, LL-37 and hBD-3 may further contribute due to chemotactic effects on neutrophils, monocytes and T cells [26, 27]. We found N. brasiliensis to exhibit complete resistance to all investigated human AMPs and to be susceptible only to bovine indolicidin. N. brasiliensis is the most frequently reported cause of progressive not cutaneous and lymphocutaneous disease. Furthermore, N. brasiliensis often causes infection in otherwise immunocompetent hosts. These clinical features are in accordance with our findings, demonstrating a complete resistance of N. brasiliensis against human epithelial, i.e. keratinocyte-derived and neutrophil-derived AMPs. N. brasiliensis is known to produce a variety of proteases [28]. To evaluate a potential resistance due to proteolytic degradation of AMPs (particularly linear α-helical LL-37), CFU-assays were conducted in the presence of protease inhibitors. However, protease inhibitors did not alter AMP-resistance in N. brasiliensis. One might speculate about species-specific variances in bacterial cell wall constituents yielding to differential nocardial AMP susceptibility/resistance [29]. Additionally, other mechanisms, i.e. active efflux by multi-drug transporters or modifications on the bacterial cell surface may confer AMP resistance. The current study revealed N. brasiliensis to be susceptible only to indolicidin, a tryptophan- and proline-rich 13 amino acid peptide of bovine neutrophils.