Pili prove pertinent to enterococcal endocarditis

Enterococcus faecalis is an important agent of endocarditis and urinary tract infections, which occur frequently in hospitals. Antimicrobial therapy is complicated by the emergence of drug-resistant strains, which contribute significantly to mortality associated with E. faecalis infection. In this issue of the JCI, Nallapareddy and colleagues report that E. faecalis produces pili on its surface and that these proteinaceous fibers are used for bacterial adherence to host tissues and for the establishment of biofilms and endocarditis (see the related article beginning on page 2799). This information may enable new vaccine strategies for the prevention of E. faecalis infections

Isolation and Characterization of a Generalized Transducing Phage for Pseudomonas aeruginosa Strains PAO1 and PA14

A temperate, type IV pilus-dependent, double-stranded DNA bacteriophage named DMS3 was isolated from a clinical strain of Pseudomonas aeruginosa. A clear-plaque variant of this bacteriophage was isolated. DMS3 is capable of mediating generalized transduction within and between P. aeruginosa strains PA14 and PAO1, thus providing a useful tool for the genetic analysis of P. aeruginosa

Sortase D Forms the Covalent Bond That Links BcpB to the Tip of Bacillus cereus Pili*

Bacillus cereus and other Gram-positive bacteria elaborate pili via a sortase D-catalyzed transpeptidation mechanism from major and minor pilin precursor substrates. After cleavage of the LPXTG sorting signal of the major pilin, BcpA, sortase D forms an amide bond between the C-terminal threonine and the amino group of lysine within the YPKN motif of another BcpA subunit. Pilus assembly terminates upon sortase A cleavage of the BcpA sorting signal, resulting in a covalent bond between BcpA and the cell wall cross-bridge. Here, we show that the IPNTG sorting signal of BcpB, the minor pilin, is cleaved by sortase D but not by sortase A. The C-terminal threonine of BcpB is amide-linked to the YPKN motif of BcpA, thereby positioning BcpB at the tip of pili. Thus, unique attributes of the sorting signals of minor pilins provide Gram-positive bacteria with a universal mechanism ordering assembly of pili

Mycobacterium tuberculosis resides in lysosome-poor monocyte-derived lung cells during chronic infection.

Mycobacterium tuberculosis (Mtb) infects lung myeloid cells, but the specific Mtb-permissive cells and host mechanisms supporting Mtb persistence during chronic infection are incompletely characterized. We report that after the development of T cell responses, CD11clo monocyte-derived cells harbor more live Mtb than alveolar macrophages (AM), neutrophils, and CD11chi monocyte-derived cells. Transcriptomic and functional studies revealed that the lysosome pathway is underexpressed in this highly permissive subset, characterized by less lysosome content, acidification, and proteolytic activity than AM, along with less nuclear TFEB, a regulator of lysosome biogenesis. Mtb infection does not drive lysosome deficiency in CD11clo monocyte-derived cells but promotes recruitment of monocytes that develop into permissive lung cells, mediated by the Mtb ESX-1 secretion system. The c-Abl tyrosine kinase inhibitor nilotinib activates TFEB and enhances lysosome functions of macrophages in vitro and in vivo, improving control of Mtb infection. Our results suggest that Mtb exploits lysosome-poor lung cells for persistence and targeting lysosome biogenesis is a potential host-directed therapy for tuberculosis

Characteristics of patients with drug susceptible, PZA resistant, and MDR mycobacterial infection in San Francisco between 1991 and 2011.

<p>Abbreviations: HIV, Human Immunodeficiency Virus; AFB, acid-fast bacilli; Ref, referent group.</p>a<p>One of the susceptible cases was Native-American.</p>b<p>Housing status was not known for one of the patients.</p>c<p>X-ray data was not available for one of the susceptible controls and one of the PZA-MDR cases.</p>d<p>Three of the susceptible cases died upon diagnosis and one moved. One of the PZA monoresistant cases died before treatment. One of the MDR cases was lost to follow up. Treatment regimens were not available for 5 of the PZA monoresistant, 7 of the MDR, and 2 of the PZA-MDR cases. These cases were excluded from treatment outcomes analysis.</p>e<p><i>p</i> values for multinomial logistical regression analysis are not recorded for categories with zero cases.</p

Site of infection based on tuberculosis drug resistance pattern in San Francisco between 1991 and 2011.

a<p>Five of the pulmonary and seven of the extrapulmonary infections were from <i>M. bovis</i>.</p>b<p><i>p</i> values for multinomial logistical regression analysis are not recorded for categories with zero cases.</p

Dynamic post-translational modification profiling of M. tuberculosis-infected primary macrophages

Macrophages are highly plastic cells with critical roles in immunity, cancer, and tissue homeostasis, but how these distinct cellular fates are triggered by environmental cues is poorly understood. To uncover how primary murine macrophages respond to bacterial pathogens, we globally assessed changes in post-translational modifications of proteins during infection with Mycobacterium tuberculosis, a notorious intracellular pathogen. We identified hundreds of dynamically regulated phosphorylation and ubiquitylation sites, indicating that dramatic remodeling of multiple host pathways, both expected and unexpected, occurred during infection. Most of these cellular changes were not captured by mRNA profiling, and included activation of ubiquitin-mediated autophagy, an evolutionarily ancient cellular antimicrobial system. This analysis also revealed that a particular autophagy receptor, TAX1BP1, mediates clearance of ubiquitylated Mtb and targets bacteria to LC3-positive phagophores. These studies provide a new resource for understanding how macrophages shape their proteome to meet the challenge of infection

Autophagy restricts Mycobacterium tuberculosis during acute infection in mice

Whether or not autophagy has a role in defence against Mycobacterium tuberculosis infection remains unresolved. Previously, conditional knockdown of the core autophagy component ATG5 in myeloid cells was reported to confer extreme susceptibility to M. tuberculosis in mice, whereas depletion of other autophagy factors had no effect on infection. We show that doubling cre gene dosage to more robustly deplete ATG16L1 or ATG7 resulted in increased M. tuberculosis growth and host susceptibility in mice, although ATG5-depleted mice are more sensitive than ATG16L1- or ATG7-depleted mice. We imaged individual macrophages infected with M. tuberculosis and identified a shift from apoptosis to rapid necrosis in autophagy-depleted cells. This effect was dependent on phagosome permeabilization by M. tuberculosis. We monitored infected cells by electron microscopy, showing that autophagy protects the host macrophage by partially reducing mycobacterial access to the cytosol. We conclude that autophagy has an important role in defence against M. tuberculosis in mammals