Smoking is associated with increased susceptibility to tuberculosis and influenza. However, little information available on the mechanisms underlying this susceptibility. Mice were left unexposed or exposed cigarette smoke then infected Mycobacterium by aerosol influenza A intranasal infection. Some mice given a DNA vaccine encoding an immunogenic M. protein. Gamma interferon (IFN-γ) production T cells from lungs spleens was measured. Cigarette exposure inhibited lung T-cell of IFN-γ during...

The continuing emergence of new strains antibiotic-resistant bacteria has renewed interest in phage therapy; however, there been limited progress applying therapy to multi-drug resistant Mycobacterium tuberculosis (Mtb) infections. In this study, we show that bacteriophage D29 and DS6A can efficiently lyse Mtb H37Rv 7H10 agar plates. However, only kills liquid culture Mtb-infected human primary macrophages. We further subsequent experiments that, after the humanized mice were infected with...

Abstract We studied the role of NK cells in regulating human CD8+ T cell effector function against mononuclear phagocytes infected with intracellular pathogen Mycobacterium tuberculosis. Depletion from PBMC healthy tuberculin reactors reduced frequency M. tuberculosis-responsive CD8+IFN-γ+ and decreased their capacity to lyse tuberculosis-infected monocytes. The CD8+IFN-γ+cells was restored by soluble factors produced activated dependent on IFN-γ, IL-15, IL-18. tuberculosis-activated...

Abstract We used human tuberculosis as a model to investigate the role of NK cytotoxic mechanisms in immune response intracellular infection. Freshly isolated cells and cell lines from healthy donors lysed Mycobacterium tuberculosis-infected monocytes greater extent than uninfected monocytes. Lysis infected was associated with increased expression mRNA for NKp46 receptor, but not NKp44 receptor. Antisera markedly inhibited lysis cell-mediated due reduced MHC class I molecules on surface or...

To determine whether the extent of spread Mycobacterium tuberculosis strains in community correlated with their capacity to replicate human macrophages, intracellular growth rates M. patient isolates were measured. Strain 210 caused disease 43 patients central Los Angeles, 3 "small-cluster" 8–23 patients, and 5 "unique" each only 1 who was positive by sputum acid-fast smear spent substantial amounts time at homeless shelters that transmission sites. grew significantly more rapidly than...

Two-component systems are important constituents of bacterial regulatory networks. Results this investigation into the role MprAB two-component system Mycobacterium tuberculosis indicate that it is associated with regulation several stress-responsive regulons. Using a deletion mutant lacking portions response regulator, MprA, and histidine kinase, MprB, was demonstrated by real-time PCR, primer extension analyses DNA microarrays activates sigma factor genes sigE sigB, under SDS stress during...

The ESX-1 secretion system exports the immunomodulatory protein ESAT-6 and other proteins important in pathogenesis of Mycobacterium tuberculosis. Components substrates are encoded at several loci, but regulation encoding genes is only partially understood. In this study, we investigated role MprAB two-component activity. We determined that directly regulates espA gene cluster, a locus necessary for function. Transcript mapping five cluster form an operon with two transcriptional start...

To evaluate the immunologic factors that contribute to protection against Mycobacterium avium complex (MAC), cytokine production by peripheral blood mononuclear cells (PBMC) from human immunodeficiency virus-negative persons with pulmonary MAC (MAC patients) and healthy control subjects a delayed hypersensitivity skin test response M. sensitin (MAS-positive subjects) was measured. In patients, mycobacterium-stimulated PBMC produced higher concentrations of interleukin (IL)-10 but lower...

We measured serum cytokine concentrations and Mycobacterium tuberculosis–stimulated production by peripheral blood mononuclear cells (PBMCs) obtained from persons infected with M. tuberculosis. Serum interferon-γ (IFN-γ) interleukin-10 (IL-10) were elevated in patients tuberculosis compared healthy who had reactions to tuberculin skin tests, but IL-18 not. In contrast, PBMCs produced less IFN-γ similar amounts of IL-10, subjects tests. Pretreatment reaction inhibited response tuberculosis,...

The Mycobacterium tuberculosis early secreted Ag of 6 kDa (ESAT-6) is a potent for human T cells and putative vaccine candidate. However, ESAT-6 also contributes to virulence in animal models, mediates cellular cytolysis, inhibits IL-12 production by mononuclear phagocytes. We evaluated the effects its molecular chaperone, culture filtrate protein 10 (CFP10), on capacity produce IFN-gamma proliferate response TCR activation. Recombinant ESAT-6, but not CFP10, markedly inhibited stimulated...

IFN-gamma production by T cells is pivotal for defense against many pathogens, and the proximal promoter of IFN-gamma, -73 to -48 bp upstream transcription start site, essential its expression. However, transcriptional regulation mechanisms through this in primary human remain unclear. We studied effects cAMP response element binding protein/activating factor (CREB/ATF) AP-1 factors on stimulated with Mycobacterium tuberculosis. Using EMSA, supershift assays, pulldown we demonstrated that...

As early secreted antigenic target of 6 kDa (ESAT-6) Mycobacterium tuberculosis (Mtb) is an essential virulence factor and macrophages are critical for infection immunity, we studied ESAT-6 stimulated IL-6 production by macrophages. significantly higher secretion murine bone marrow derived (BMDM) compared to culture filtrate protein 10 (CFP10) antigen 85A. Polymyxin B, LPS blocker, did not affect macrophage production. but Pam3CSK4 induced TLR2 knockout BMDM. phosphorylation DNA binding...

The goal of this study was to understand the role altered mitochondrial function in breast cancer progression and determine potential molecular alteration signature developing exosome-based biomarkers.This designed characterize critical components regulating tumorigenesis. Experiments were conducted assess these molecules for biomarker development.We observed a remarkable reduction spontaneous metastases through interplay mitochondria by SH3GL2, vesicular endocytosis-associated protein MFN2,...

Summary The mobile insertion sequence, IS 6110 , is an important marker in tracking of Mycobacterium tuberculosis strains. Here, we demonstrate that can upregulate downstream genes through outward‐directed promoter its 3′ end, thus adding to the significance this element. Promoter activity was orientation dependent and localized within a 110 bp fragment adjacent right terminal inverted repeat. Transcripts from promoter, named OP6110, begin ≈ 85 upstream end . Use green fluorescent protein...

Early secreted antigenic target of 6 kDa (ESAT-6) Mycobacterium tuberculosis is critical for the virulence and pathogenicity M. tuberculosis. IL-8, a major chemotactic cytokine neutrophils T lymphocytes, plays important roles in development lung injury. To further understand role ESAT-6 pathology associated with development, we studied effects on regulation IL-8 expression epithelial cells. induced by increasing gene transcription mRNA stability. induction promoter activity was dependent...

Previously, we found that pathological immune responses enhance the mortality rate of Mycobacterium tuberculosis (Mtb)-infected mice with type 2 diabetes mellitus (T2DM). In current study, evaluated role cytokine IL-22 (known to play a protective in bacterial infections) and 3 innate lymphoid cells (ILC3s) regulating inflammation Mtb-infected T2DM mice. levels were significantly lower than nondiabetic Similarly, serum (TB) patients TB without T2DM. ILC3s an important source infected Mtb,...

Programmed cell death and especially necroptosis, a programmed regulated form of necrosis, have been recently implicated in the progression outcomes influenza mouse models. Moreover, Z-DNA/RNA binding protein 1 (ZBP1) has identified as key signaling molecule for necroptosis induced by A virus (IAV). Direct evidence IAV-induced not shown infected lungs vivo. It is also unclear to what types undergo during pulmonary IAV infection whether ZBP1 expression can be inflammatory mediators. In this...

Summary The ability of Mycobacterium tuberculosis to grow in macrophages is central its pathogenicity. We found previously that the widespread 210 strain M. grew more rapidly than other strains human macrophages. Because principal sigma factors influence virulence some bacteria, we analysed mRNA expression factor, sigA , isolates during growth Isolates had higher levels and intracellular rates, compared with clinical laboratory H37Rv. SigA was also upregulated isolate TB294 macrophages,...

To investigate the role of interleukin (IL)-18 in human tuberculosis, IL-18 production was evaluated blood and at site disease patients with tuberculosis. Mycobacterium tuberculosis-stimulated peripheral mononuclear cells (PBMC) from tuberculosis secreted less interferon-γ (IFN-γ) than did PBMC healthy persons reactive to tuberculin. M. tuberculosis-induced IFN-γ inhibited by anti-IL-18 enhanced recombinant IL-18. Alveolar macrophages response concentrations were higher pleural fluid...

We reported previously that the early secreted antigenic target of 6 kDa (ESAT-6) from Mycobacterium tuberculosis directly inhibits human T cell IFN-γ production and proliferation in response to stimulation with anti-CD3 anti-CD28. To determine mechanism this effect, we treated cells kinase inhibitors before ESAT-6. Only p38 MAPK inhibitor, SB203580, abrogated ESAT-6-mediated inhibition a dose-dependent manner. SB203580 did not reverse IL-17 IL-10 production, suggesting specific effect on...

Early secreted antigenic target of 6 kDa (ESAT-6) Mycobacterium tuberculosis is a T cell Ag that potential vaccine candidate, but it also virulence factor mediates pathogenicity. To better understand the effects ESAT-6 on immune response, we studied effect human dendritic cells (DCs). Peripheral blood monocytes were treated with GM-CSF and IL-4 to yield immature DCs, which matured by addition LPS CD40 ligand (CD40L), or without ESAT-6. inhibited LPS/CD40L-induced DC expression costimulatory...

The regulatory mechanisms that control the ESX-1 secretion system, a key player in pathogenesis of Mycobacterium tuberculosis, have not been fully elucidated. However, factors regulate substrate EspA usually affect function. Previous studies showed espA is directly regulated by nucleoid-associated protein EspR and two-component system (TCS) MprAB. PhoPR TCS also activates espA, but direct target PhoP was unknown. In this report, we reveal MprA PhoP-Rv, PhoP-Ra. PhoP-Rv binding sites espR...

Abstract Macrophages are professional phagocytes known to play a vital role in controlling Mycobacterium tuberculosis ( Mtb ) infection and disease progression. Here we compare growth mouse alveolar (AMs), peritoneal (PMs), liver (Kupffer cells; KCs) macrophages bone marrow-derived monocytes (BDMs). KCs restrict more efficiently than all other despite equivalent infections through enhanced autophagy. A metabolomics comparison of -infected indicates that ornithine imidazole two top-scoring...

ABSTRACT A major obstacle to development of subunit vaccines and diagnostic reagents for tuberculosis is the inability produce large quantities these proteins. To test hypothesis that poor expression some mycobacterial genes in Escherichia coli due, part, presence low-usage E. codons, we used site-directed mutagenesis convert codons high-usage same amino acid Mycobacterium antigens 85A 85B superoxide dismutase. Replacement five wild-type gene antigen increased recombinant protein production...