The C terminus of the mycobacterium ESX-1 secretion system substrate ESAT-6 is required for phagosomal membrane damage and virulence.

SignificanceTuberculosis (TB), an ancient disease of humanity, continues to be a major cause of worldwide death. The causative agent of TB, Mycobacterium tuberculosis, and its close pathogenic relative Mycobacterium marinum, initially infect, evade, and exploit macrophages, a major host defense against invading pathogens. Within macrophages, mycobacteria reside within host membrane-bound compartments called phagosomes. Mycobacterium-induced damage of the phagosomal membranes is integral to pathogenesis, and this activity has been attributed to the specialized mycobacterial secretion system ESX-1, and particularly to ESAT-6, its major secreted protein. Here, we show that the integrity of the unstructured ESAT-6 C terminus is required for macrophage phagosomal damage, granuloma formation, and virulence.

Analysis of the Diversity and Functional Potential of Bacterial Communities in Tuberculomas.

The dynamics of lung microbiota in tuberculosis remains poorly understood. Sequencing of variable regions of the 16S rRNA gene from surgically excised tuberculosis foci and biopsy specimens of normal lung tissue allowed characterization of the diversity and predictive potential of bacterial communities. Taxonomic diversity indices attested to differences in the structure of microbial communities between "healthy" lungs and tuberculomas. The microbial composition of "healthy" lungs varied in taxonomic diversity and was presented by both gram-positive and gram-negative bacteria with sufficiently similar metabolic potential. The microbiota of the examined tuberculomas consisted of Mycobacterium tuberculosis in 99.9% of cases. A significant part of the metabolic pathways predicted by PICRUSt2 included cholesterol catabolism, sulfate assimilation, and various pathways for the biosynthesis of cell wall components.

Diagnosis and treatment of paediatric tuberculosis: An insight review.

Tuberculosis (TB) is a major public health problem, invading all age groups world-wide. It is an opportunistic infection affecting the individuals alone or with co-infections. Childhood TB is a neglected aspect and a significant health problem in epidemic areas. It constitutes more than 20% of TB incidence. Pediatric TB exists in the shadow of adult TB. The clinicians concentrate on pulmonary manifestation of TB, whereas it is a major problem in both pulmonary and extra-pulmonary infections. The rate of infection with this disease is mostly associated with poverty, social disruption and human immunodeficiency virus (HIV) infection. The diagnosis of extra-pulmonary TB (EPTB) is more difficult than pulmonary TB (PTB). Delayed diagnosis and executive treatment contribute to increase in the mortality rate in endemic areas. This article provides the evidence-based simple and safe screening method, indicating rapid, highly sensitive and specific diagnostic tests for pulmonary and EPTB in children. The most important aspect of treatment is the correct course of anti-tubercular drugs. This review serves the purpose of quick reference for microbiologists, epidemiologists, academicians, students and researchers. It provides guidance regarding early diagnosis and treatment accuracy of pediatric TB.

Modeling nanoparticle delivery of TB drugs to granulomas.

Tuberculosis, which typically presents as a pulmonary disease, has a complex pathology. The primary site of infection, the Ghon focus, recruits immune cells and a granuloma forms. At earlier stages the granuloma is still vascularized, offering the best opportunity for drug treatment. In the more progressive state blood flow is reduced and a distinct caseous structure develops. Effective delivery of drugs to bacilli in the core of the granuloma becomes very difficult. It is perceivable that granuloma cores could create conditions where bacilli persist and develop resistance. In this study we analyze drug delivery to granulomas by means of a nanoparticle delivery system. The model consists of two parts; the overall distribution of the nanoparticles is described by a simple circulatory model and this result is used in the second part, focusing on transport in a capillary lined with macrophages. Nanoparticles enter the macrophages where they are metabolized and the drugs are released. The model reveals significant differences in drug concentrations between the plasma and macrophages. Based on the results of the model, strategies for improved drug delivery are proposed.

Intracardiac tuberculomas caused by Mycobacterium tuberculosis in a dog.

BACKGROUND: This paper presents an unusual form of disseminated Mycobacterium tuberculosis infection in a dog. The infection lasted at least one year and its main gross lesions were massive cardiac tuberculomas. To the best of our knowledge, this is the first report of heart tuberculomas in a dog. CASE PRESENTATION: A 9-year-old mixed-breed male dog weighing 10 kg was referred to the clinic for cardiological evaluation before general anesthesia. The echocardiography revealed a lump of about 20 mm in diameter in the area of the left atrium. Almost one year later the same dog was presented again in severe clinical state (fever, anorexia, weight loss, depression, cough, dyspnea, lymphadenomegaly, vomiting, recent episodes of fainting). Due to progression of the disease and poor effects of treatment the owner decided to euthanize the dog. Most prominent lesions observed during autopsy were diffuse pneumonia, fibrinous pericarditis and epicarditis as well as large, yellow, semisolid masses of caseous necrosis in the left and right atrium (30 mm and 15 mm in diameter, respectively). From both pulmonary and cardiac lesions M. tuberculosis was isolated on Lowenstein-Jensen slants and in Bactec Mycobacteria Growth Indicator Tube 960 liquid media, and confirmed by BD ProbeTec ET Direct Detection Assay and spoligotyping. CONCLUSION: Companion animals may occasionally suffer from tuberculosis but majority of cases probably remain misdiagnosed or undetected. Typically tuberculosis in dogs affects lungs and their regional lymph nodes. Even in humans tuberculomas are rare manifestation of mycobacterial infection, mostly seen in the central nervous system. Atypical location of main tuberculous lesions may account for lack of correct ante mortem diagnosis in this case.

Mycobacterium tuberculosis: Manipulator of Protective Immunity.

Mycobacterium tuberculosis (MTB) is one of the most successful pathogens in human history and remains a global health challenge. MTB has evolved a plethora of strategies to evade the immune response sufficiently to survive within the macrophage in a bacterial-immunological equilibrium, yet causes sufficient immunopathology to facilitate its transmission. This review highlights MTB as the driver of disease pathogenesis and presents evidence of the mechanisms by which MTB manipulates the protective immune response into a pathological productive infection.

[Interlaboratory test: Isolation of Mycobacterium bovis from granulomatous lesions in bovine]. / Ensayo interlaboratorio: aislamiento de Mycobacterium bovis a partir de lesiones granulomatosas en bovinos.

Mycobacterium bovis is the causative agent of bovine tuberculosis. The diagnostic laboratory confirmation is made through bacterial isolation. The aim of interlaboratory tests is to assess the performance of each participant in comparison with other of similar capacities. The test objective was to determine the efficiency of isolation of M. bovis. Four laboratories were part of the test and processed 25 blind tissue samples from granulomatous lesions and with previous M. bovis isolation. The laboratory that had the highest proportion of isolates was A (68%), followed by C (60%) and then B and D (both with 52%). The greatest concordance was observed between B-D and B-C laboratories (68%). The differences could be due to specific factors in each laboratory procedures. This type of interlaboratory tests highlights errors in the bacteriology and identifies critical points in the process to detect M. bovis accurately.

Differential risk of tuberculosis reactivation among anti-TNF therapies is due to drug binding kinetics and permeability.

Increased rates of tuberculosis (TB) reactivation have been reported in humans treated with TNF-α (TNF)-neutralizing drugs, and higher rates are observed with anti-TNF Abs (e.g., infliximab) as compared with TNF receptor fusion protein (etanercept). Mechanisms driving differential reactivation rates and differences in drug action are not known. We use a computational model of a TB granuloma formation that includes TNF/TNF receptor dynamics to elucidate these mechanisms. Our analyses yield three important insights. First, drug binding to membrane-bound TNF critically impairs granuloma function. Second, a higher risk of reactivation induced from Ab-type treatments is primarily due to differences in TNF/drug binding kinetics and permeability. Apoptotic and cytolytic activities of Abs and pharmacokinetic fluctuations in blood concentration of drug are not essential to inducing TB reactivation. Third, we predict specific host factors that, if augmented, would improve granuloma function during anti-TNF therapy. Our findings have implications for the development of safer anti-TNF drugs to treat inflammatory diseases.

Looking within the zebrafish to understand the tuberculous granuloma.

Tuberculosis is characterized by the formation of complex immune cell aggregates called granulomas, which for nearly a century have been viewed as critical host-beneficial structures to restrict bacterial growth and spread. A different view has now emerged from real-time visualization of granuloma formation and its consequences in the optically transparent and genetically tractable zebrafish larva. Pathogenic mycobacteria have developed mechanisms to use host granulomas for their expansion and dissemination, at least during the innate phases of infection. Host processes that are intended to be beneficial-death of infected macrophages and their subsequent phagocytosis by macrophages that are newly recruited to the growing granuloma-are harnessed by mycobacteria for their own benefit. Mycobacteria can also render the granuloma a safe-haven in the more advanced stages of infection. An understanding of the host and bacterial pathways involved in tuberculous granuloma formation may suggest new ways to combat mycobacterial infection.

Primary musculoskeletal mycobacterium infection with large cystic masses after total hip arthroplasty.

Primary mycobacterial infections in the musculoskeletal system are rare with a limited number of published case reports. This report describes a case involving a primary musculoskeletal tuberculous abscess. A 62-year-old male patient who had a right total hip arthroplasty performed 8 years earlier, using metal-on-metal articulation presented with a 1-year history of non-tender masses on his right thigh. Initially, it was assumed he had metallosis. Intraoperatively, an incision into the mass was conducted which resulted in draining of a whitish-grey pus like fluid. A diagnosis of tuberculosis was confirmed with both microscopic and histological examination. The patient was treated over a course of six months with an anti-tuberculosis medication regimen following the confirmation of a solitary soft tissue tuberculosis infection. At the 24 month follow-up, the patient was asymptomatic with no relapse of the mass.

Metabolomic signatures in guinea pigs infected with epidemic-associated W-Beijing strains of Mycobacterium tuberculosis.

With the understanding that the laboratory propagated strain of Mycobacterium tuberculosis H37Rv is of modest virulence and is drug susceptible, in the present study, we performed a nuclear magnetic resonance-based metabolomic analysis of lung tissues and serum obtained from guinea pigs infected by low dose aerosol exposure to clinical isolates of Mycobacterium tuberculosis. High Resolution Magic Angle Spinning NMR coupled with multivariate statistical analysis of 159 lung tissues obtained from multiple locations of age-matched naïve and 30 and 60 days of infected guinea pig lungs revealed a wide dispersal of metabolic patterns, but within these, distinct clusters of signatures could be seen that differentiated between naive control and infected animals. Several metabolites were identified that changed in concert with the progression of each infection. Major metabolites that could be interpreted as indicating host glutaminolysis were consistent with activated host immune cells encountering increasingly hypoxic conditions in the necrotic lung lesions. Moreover, glutathione levels were constantly elevated, probably in response to oxygen radical production in these lesions. Additional distinct signatures were also seen in infected serum, with altered levels of several metabolites. Multivariate statistical analysis clearly differentiated the infected from the uninfected sera; in addition, Receiver Operator Characteristic curve generated with principal component 1 scores showed an area under the curve of 0.908. These data raise optimism that discrete metabolomic signatures can be defined that can predict the progression of the tuberculosis disease process, and form the basis of an innovative and rapid diagnostic process.

Host-detrimental role of Esx-1-mediated inflammasome activation in mycobacterial infection.

The Esx-1 (type VII) secretion system is a major virulence determinant of pathogenic mycobacteria, including Mycobacterium marinum. However, the molecular events and host-pathogen interactions underlying Esx-1-mediated virulence in vivo remain unclear. Here we address this problem in a non-lethal mouse model of M. marinum infection that allows detailed quantitative analysis of disease progression. M. marinum established local infection in mouse tails, with Esx-1-dependent formation of caseating granulomas similar to those formed in human tuberculosis, and bone deterioration reminiscent of skeletal tuberculosis. Analysis of tails infected with wild type or Esx-1-deficient bacteria showed that Esx-1 enhanced generation of proinflammatory cytokines, including the secreted form of IL-1beta, suggesting that Esx-1 promotes inflammasome activation in vivo. In vitro experiments indicated that Esx-1-dependent inflammasome activation required the host NLRP3 and ASC proteins. Infection of wild type and ASC-deficient mice demonstrated that Esx-1-dependent inflammasome activation exacerbated disease without restricting bacterial growth, indicating a host-detrimental role of this inflammatory pathway in mycobacterial infection. These findings define an immunoregulatory role for Esx-1 in a specific host-pathogen interaction in vivo, and indicate that the Esx-1 secretion system promotes disease and inflammation through its ability to activate the inflammasome.

Cystic pulmonary tuberculoma.

Pulmonary tuberculosis (TB) is a medical and social problem, particularly in developing countries. Early diagnosis and treatment is important. Chest radiography is usually the first diagnostic tool when there is a suspicion of pulmonary TB. A computed tomography (CT) scan provides more accurate information on the extent and distribution of pulmonary TB. We present here a young, immunocompetent male patient with unusual imaging findings for pulmonary TB. We discuss the clinical presentation and management.

The secret trumps, impelling the pathogenicity of tubercle bacilli.

Confrontation between invading microbial pathogens and host defense systems involves intricate cellular and molecular interactions. Here we discuss the virulence factors as trumps, overriding the contest in favor of the tubercle bacillus (Mycobacterium tuberculosis). It evolved a number of molecular constituents, which can interfere with antigen presentation and Toll receptor function, thus impairing immune defenses. It also evolved stress responses, which can drive its cell cycle into a non-replicating, low metabolic mode. Although the low counts of latent bacilli prevent their direct detection, we contend that they retain a capacity to survive for long periods in foamy macrophages and within the necrotic parts of lung granulomas. We attributed significance to drainage of M. tuberculosis by the alveolar fluid: while out-flow is responsible for the clearance, the reverse-flow has an important capacity to re-infect the lungs and to transmit the infection to new recipients. We consider the cycling between replicating and latent organisms to be a continuous process, which is a departure from the concept of long-lived dormant organisms, with a capacity to resuscitate. These aspects impinge also on the actions of isoniazid (INH) chemotherapy and on the topography of human lung lesions. Eventually, fibrosis of the connective tissue of the lungs is known to encapsulate lung lesions, thus limiting the impact of both outward and reverse drainage. In conclusion, the novelty of our views on M. tuberculosis-host interactions rests in the dynamic perception of M. tuberculosis latency and its evolutionary importance for the pathogenesis of tuberculosis.

Solitary pulmonary nodules caused by Mycobacterium tuberculosis and Mycobacterium avium complex.

Few studies have compared the clinical and radiographic findings of tuberculomas to those of solitary pulmonary nodules (SPNs) caused by Mycobacterium avium complex (MAC). We retrospectively analyzed clinical and radiographic findings from 26 patients with tuberculomas and 15 patients with SPNs caused by MAC. Median SPN size was 22 mm. In 26 patients (63%), the SPN was detected during a routine health checkup or evaluation of organs other than lungs. Patients with SPNs due to MAC were slightly older (median = 59 years) compared with those with tuberculomas (median = 50 years; P = 0.044). When we compared computed tomography (CT) features between patients with tuberculomas and patients with MAC, no significant differences were found in SPN location or the presence of calcification, cavitation, central low attenuation, and the satellite lesions. Although the maximum standardized uptake values were slightly higher in patients with SPNs due to MAC (median = 8.5) compared with those with tuberculomas (median = 2.2), this difference was not significant (P = 0.053). Of the 15 patients with SPNs due to MAC, 10 were initially diagnosed with "tuberculoma" and administered antituberculosis medication. MAC pulmonary disease should be considered in the differential diagnosis of SPNs, even when encountered in geographic regions with a high prevalence of pulmonary tuberculosis.

[Primary pancreatic tuberculosis in an immunocompetent patient: first case report in Spain]. / Tuberculosis pancreática primaria en un paciente inmunocompetente: primer caso comunicado en España.

Primary pancreatic tuberculosis (PPTB) is an extremely rare entity defined by an isolated pancreatic lesion with microbiological confirmation, in the absence of previously identified tuberculosis (TB) and involvement of any other organ. We report the case of a 47-year-old man referred for abdominal pain and weight loss, in whom several imaging techniques revealed a solid mass in the head of the pancreas. CT-guided fine-needle aspiration cytology was consistent with necrotic granuloma. Intradermic tuberculin reaction was positive, but acid fast bacilli staining was negative in repeat cytology. No additional evidence of TB was found after exhaustive diagnostic work-up. Exploratory laparotomy was proposed for a definitive diagnosis, but cultures grew Mycobacterium tuberculosis at 50 days. The pancreatic lesion disappeared after 4 months of antitubercular therapy. This is the first case report of PPTB in an immunocompetent person in Spain. A high index of suspicion and accurate samples for microbiology are mandatory to avoid unnecessary surgical procedures.

Resolution of Large Choroidal Tuberculoma following Monotherapy with Intravitreal Ranibizumab.

Background: Ocular tuberculosis can have protean manifestations. Anti-tubercular therapy (ATT) and oral steroids are employed in the management of this condition. There is evidence in the literature which has highlighted the use of intravitreal anti-vascular endothelial growth factor drugs as an adjunct to systemic therapy.Report of the Case: A 44-year-old male presented with a decrease of vision in the right eye was diagnosed choroidal tuberculoma with massive exudation and subretinal fluid. The patient was treated with intravitreal ranibizumab injection. The lesion regressed completely within 6 weeks without any additional systemic corticosteroids and ATT without any recurrence over 6 months during follow-up.Conclusions: Ranibizumab monotherapy may lead in complete regression of vascularized tubercular choroidal granulomas without the need of adjunctive ATT and corticosteroids. After intravitreal injection of ranibizumab, the lesion may be observed for regression over several weeks.

Paradigm changing evidence that alter tuberculosis perception and detection: Focus on latency.

Tuberculosis remains a devastating disease to Mankind, ranking as the ninth cause of death worldwide. Eliminating tuberculosis as proven much more difficult than once anticipated. In addition to the delay in diagnosis and drug resistance problems that compromise the efficacy of treatment, the enormous reservoir of latently infected individuals continuously feeds the epidemics. However, targeting latency with prophylactic antibiotic administration is not possible at the populational level. Together, these issues call for a better understanding of latency, as well as for a more precise identification of individuals at high risk of reactivation. For this, recent paradigm changing evidence need to be taken into account, most notably, the existence of a tuberculosis spectrum; the genetic diversity of both humans and tuberculosis-causing bacteria; and the changes in the human population that interfere with tuberculosis. Here we discuss latency in the light of these variables and how that understanding can move forward tuberculosis research and elimination.