Chronic TB Research Articles

Change in smoking behaviour of people who suffer from respiratory disease.

OBJECTIVE: To analyse smoking behaviour in patients suffering from non-small cell lung cancer (NSCLC), chronic obstructive pulmonary disease (COPD) and pulmonary TB (PTB).METHODS: The study population comprised 421 inpatients newly diagnosed with NSCLC, COPD and PTB at the First Affiliated Hospital of Chongqing Medical University, Chongqing, China, and 362 healthy individuals (controls) recruited from September 2016 to March 2017. All participants were current smokers. After enrolment, the subjects were invited to participate in face-to-face interviews to complete the self-designed questionnaire. A follow-up survey was performed 2 years later.RESULTS: Two years after being diagnosed, patients with NSCLC, COPD and PTB had smoking cessation rates of respectively 76.8%, 62.8% and 63.7%. The cessation rate was only 10.2% in the control group (P < 0.01). The difficulty of smoking cessation after diagnosis in patients with the three diseases was significantly decreased (P < 0.01).CONCLUSION: The smoking cessation rate among participants suffering from NSCLC, COPD and PTB increased after diagnosis compared with that before diagnosis, and was significantly higher than the control group of healthy individuals.

S100A8/A9 regulates CD11b expression and neutrophil recruitment during chronic tuberculosis.

Neutrophil accumulation is associated with lung pathology during active tuberculosis (ATB). However, the molecular mechanism or mechanisms by which neutrophils accumulate in the lung and contribute to TB immunopathology are not fully delineated. Using the well-established mouse model of TB, our new data provide evidence that the alarmin S100A8/A9 mediates neutrophil accumulation during progression to chronic TB. Depletion of neutrophils or S100A8/A9 deficiency resulted in improved Mycobacterium tuberculosis (Mtb) control during chronic but not acute TB. Mechanistically, we demonstrate that, following Mtb infection, S100A8/A9 expression is required for upregulation of the integrin molecule CD11b specifically on neutrophils, mediating their accumulation during chronic TB disease. These findings are further substantiated by increased expression of S100A8 and S100A9 mRNA in whole blood in human TB progressors when compared with nonprogressors and rapidly decreased S100A8/A9 protein levels in the serum upon TB treatment. Furthermore, we demonstrate that S100A8/A9 serum levels along with chemokines are useful in distinguishing between ATB and asymptomatic Mtb-infected latent individuals. Thus, our results support targeting S100A8/A9 pathways as host-directed therapy for TB.

Intracellular and in vivo evaluation of imidazo[2,1-b]thiazole-5-carboxamide anti-tuberculosis compounds.

The imidazo[2,1-b]thiazole-5-carboxamides (ITAs) are a promising class of anti-tuberculosis agents shown to have potent activity in vitro and to target QcrB, a key component of the mycobacterial cytochrome bcc-aa3 super complex critical for the electron transport chain. Herein we report the intracellular macrophage potency of nine diverse ITA analogs with MIC values ranging from 0.0625–2.5 μM and mono-drug resistant potency ranging from 0.0017 to 7 μM. The in vitro ADME properties (protein binding, CaCo-2, human microsomal stability and CYP450 inhibition) were determined for an outstanding compound of the series, ND-11543. ND-11543 was tolerable at >500 mg/kg in mice and at a dose of 200 mg/kg displayed good drug exposure in mice with an AUC(0-24h) >11,700 ng·hr/mL and a >24 hr half-life. Consistent with the phenotype observed with other QcrB inhibitors, compound ND-11543 showed efficacy in a chronic murine TB infection model when dosed at 200 mg/kg for 4 weeks. The efficacy was not dependent upon exposure, as pre-treatment with a known CYP450-inhibitor did not substantially improve efficacy. The ITAs are an interesting scaffold for the development of new anti-TB drugs especially in combination therapy based on their favorable properties and novel mechanism of action.

P1.16-16 Lung Cancer Control in Sub-Saharan Africa: Experience at Ampath Oncology in Western Kenya

Objective To find out the cause for the under-diagnosis of Lung Cancer (LC) at AMPATH by using community engagement and high-risk screening at the TB clinics. Methods FGDs with community cough monitors in counties were done due to overlap of LC and TB presentations. Consequently, through establishing a multinational-lung cancer control program (MLCCP) to improve diagnosis and patient journey for lung cancer patients in our settings, we classified patients with symptomatic lung disease (chest pain, cough, SOB, weight loss, hemoptysis) and negative gene expert/negative sputum for AAFB as high-risk for further evaluation. CT scans were done for anyone with a chest mass/lesion and Image-guided biopsy offered. s Jan 2018-Mar 2019, 331 high risk clients were evaluated. 214 with masses CT scans of which 205 were lung and 9 were mediastinal. 131/214 had biopsy, of which 83 (60 LC, 23 secondary mets) while 48 were other conditions. These included: Lung Fibrosis, Aspergillosis, Chronic granulomatous inflammation, TB, Thymoma, viral histiocytosis, Granuloma and unconfirmed diagnosis For the biopsied lung masses-131/214, 60 had confirmed LC. This represented 45.8% of those biopsied. Male to Female ratio was 1:1, median age at diagnosis was 62 with 55-74 age range accounting for 73.2% of LC cases. The mean duration of symptoms was 8 months, range of 1 to 12 months. >50% of the cancer patients made 7-10 hospital visits before diagnosis, with 25% making more than 14 visits. NSCLC accounted for 92.2% of the diagnosis with SCLC 7.8%. Adenocarcinoma was the commonest diagnosed histological sub-type at 66% of NSCLC. Majority of the patients were diagnosed at stage IV, 78.1% with only three patients diagnosed in stage II. 39% (25/64) patients are alive and on follow-up. Early detection is key. Poor referral patterns and lack of LC knowledge and diagnostic skills by HC professionals causes late stage at diagnosis. Patients do not present Late. Community engagement and embedding simple protocols for prompt referrals/diagnostic work-up in TB control programs may lead to improved outcomes. Prevention measures should also be rolled out. Cough monitors were essential to improving the LC patient's journey. *MLCCP is a Multi-National Lung Cancer Control Program with Dr. Asirwa the overall PI for Kenya, Tanzania, Swaziland and South Africa. Funding for the program has been provided by Bristol Myers Squib Foundation (BMSF) *MLCCP Team is the Kenyan Team for this Western Kenya Program Component

Evolution of rifampicin treatment for tuberculosis

Rifampicin was discovered in 1965 and remains one of the most important drugs in tuberculosis treatment that is valued for its sterilizing activity and ability to shorten treatment. Antimicrobial activity of rifampicin was initially proved in vitro; subsequently numerous in vivo studies showed the bactericidal properties and dose-dependent effect of rifampicin. Rifampicin was first during the late 1960s to treat patients suffering from chronic drug-resistant pulmonary TB. Decades later, rifampicin continues to be studied with particular emphasis on whether higher doses could shorten the duration of treatment without increasing relapse or having adverse effects. Lesion-specific drug penetration and pharmacokinetics of rifampicin are improving our understanding of effective concentration while potentially refining drug regimen designs. Another prospective aspect of high-dose rifampicin is its potential use in treating discrepant mutation thereby eliminating the need for MDR treatment. To date, several clinical trials have shown the safety, efficacy, and tolerability of high-dose rifampicin. Currently, high-dose rifampicin has been used successfully in a routine clinical setting for the treatment of high-risk patients. However, the WHO and other relevant policy makers have not committed to implementing a controlled rollout thereof. This review describes the course that rifampicin has travelled to the present-day exploration of high-dose rifampicin treatment.

Diabetic trends and associated mortality in tuberculosis patients in Texas, a large population-based analysis

BackgroundTuberculosis (TB) and diabetes mellitus (DM) comorbidity (TB-DM) is a major public health challenge worldwide. This analysis aimed to determine the risk factors and trends associated with TB-DM morbidity and mortality. MethodsRisk factors for TB-DM morbidity and mortality were identified by logistic regression using de-identified surveillance data of all TB patients from Texas, USA. reported between 01/2010–12/2016. Non-parametric testing was used for the morbidity and mortality trends. ResultsFrom 2010 to 2016, 1400/9002 (15.6%) TB patients were diabetic with an annual prevalence increase from 12.5% to 18.7% (p = 0.005). Reported TB-DM patients had a higher mortality (10.3%) than non-DM patients (7.6%, p = 0.001) with nearly a 3-fold increase in the odds of death (overall and during treatment). Older age, being Hispanic, chronic kidney failure, pulmonary cavitation and positive TB culture or smear were associated with TB-DM. Age ≥45, US-birth, resident of long-term care facility, injecting-drug user, chronic kidney disease, TB meningitis, abnormal chest radiograph, non-conversion of culture, and HIV(+) were independently associated with a higher mortality. ConclusionsTB-DM is an increasing public health problem in Texas with significantly high mortality. Risk factors for mortality determined by multivariate modeling will provide a foundation for the development of more effective strategies for TB-DM management.

Health seeking for chronic lung disease in central Malawi: Adapting existing models using insights from a qualitative study.

BackgroundChronic lung diseases contribute to the growing non-communicable disease (NCD) burden and are increasing, particularly in many low and middle-income countries (LMIC) in sub-Saharan African. Early engagement with health systems in chronic lung disease management is critical to maintain quality of life and prevent further damage. Our study sought to understand health seeking behaviour in relation to chronic lung disease and TB in a rural district in Malawi.MethodsQualitative data was collected between March-May 2015, exploring patterns of health seeking for lung disease amongst residents of two districts in rural Malawi. Participants included those with and without lung disease, health workers and village leaders. Participants with a history of TB were included in the sample due to similarities in clinical presentation and in view of potential to cause long-term damage to lung tissue.ResultsOur findings are ordered around a specific model of health seeking devised by adapting previous models. The model and findings span three broad areas that were found to influence health seeking: understandings of health and disease which shaped whether, when and where to seek care; the care seeking decision which was influenced by social and structural factors; and the care seeking experience which impacted future care decisions creating ‘feedback loops’.DiscussionEfforts to improve effective and accessible healthcare provision for chronic lung disease need to address all the determinants of health seeking behaviour identified. This may include: enhancing the structural and financial accessibility of health services, through the strengthening of community linkages; improving communication between formal health providers, patients and communities around symptoms, diagnosis and management of chronic lung diseases; and improving the quality of diagnostic and management services through the strengthening of health systems ‘hardware’ (equipment availability) and ‘software’ (development of trusting and respectful relationships between providers and patients).

A new model for chronic and reactivation tuberculosis: Infection with genetically attenuated Mycobacterium tuberculosis in mice with polar susceptibility

TB infection in mice develops relatively rapidly which interferes with experimental dissection of immune responses and lung pathology features that differ between genetically susceptible and resistant hosts. Earlier we have shown that the M. tuberculosis strain lacking four of five Rpf genes (ΔACDE) is seriously attenuated for growth in vivo. Using this strain, we assessed key parameters of lung pathology, immune and inflammatory responses in chronic and reactivation TB infections in highly susceptible I/St and more resistant B6 mice. ΔACDE mycobacteria progressively multiplied only in I/St lungs, whilst in B6 lung CFU counts decreased with time. Condensed TB foci apeared in B6 lungs at week 4 of infection, whilst in I/St their formation was delayed. At the late phase of infection, in I/St lungs TB foci fused resulting in extensive pneumonia, whereas in B6 lungs pathology was limited to condensed foci. Macrophage and neutrophil populations characteristically differed between I/St and B6 mice at early and late stages of infection: more neutrophils accumulated in I/St and more macrophages in B6 lungs. The expression level of chemokine genes involved in neutrophil influx was higher in I/St compared to B6 lungs. B6 lung cells produced more IFN-γ, IL-6 and IL-11 at the early and late phases of infection. Overall, using a new mouse model of slow TB progression, we demonstrate two important features of ineffective infection control underlined by shifts in lung inflammation: delay in early granuloma formation and fusion of granulomas resulting in consolidated pneumonia late in the infectious course.

Albumin Fusion with Granulocyte-Macrophage Colony-Stimulating Factor Acts as an in Situ Therapeutic Vaccine Against Chronic Tuberculosis in Mice

Background: The long duration of treatment and emerging drug resistance pose significant challenges for global tuberculosis eradication efforts. Therefore, novel strategies to shorten TB treatment regimens are urgently needed. Methods: Using an albumin fusion strategy, we created an albumin-fused granulocyte-macrophage colony stimulating factor (named albGM-CSF) novel molecule that harnesses albumin's long-half life and targeting abilities to enhance the biostability of GM-CSF and direct it to the lymph nodes where the effects of GM-CSF can increase dendritic cell populations crucial to eliciting a potent immune response. Findings: In our study, we found that subcutaneous administration of albGM-CSF reduces the mean lung bacillary burden in mice chronically-infected with tuberculosis. This effect was dependent on both innate immunity and CD4 T cells. GM-CSF induced IL-1β release from Mtb-infected dendritic cells and macrophages, and higher IL-1β levels were observed in albGM-CSF-treated mice with chronic TB infection. Interpretation: Albumin fusion with GM-CSF represents a promising tissue-targeting strategy for controlling chronic lung TB infections and serves as a novel in situ therapeutic vaccination platform for other infectious diseases and malignancies. Funding Statement: This research was supported by the National Institute of Allergy and Infectious Diseases of the National Institutes of Health under award number R21AI22922. Declarations of Interest: The authors declare no conflict of interest. Ethics Approval Statement: All procedures were performed according to the Johns Hopkins Institutional Animal Care and Use Committee and in accordance with recommendations for proper use and care of laboratory animals.

Delamanid Kills Dormant Mycobacteria In Vitro and in a Guinea Pig Model of Tuberculosis.

Tuberculosis (TB) treatment is long and requires multiple drugs, likely due to various phenotypes of TB bacilli with variable drug susceptibilities. Drugs with broad activity are urgently needed. This study aimed to evaluate delamanid's activity against growing or dormant bacilli in vitro as well as in vivo Cultures of Mycobacterium bovis BCG Tokyo under aerobic and anaerobic conditions were used to study the activity of delamanid against growing and dormant bacilli, respectively. Delamanid exhibited significant bactericidal activity against replicating and dormant bacilli at or above concentrations of 0.016 and 0.4 mg/liter, respectively. To evaluate delamanid's antituberculosis activity in vivo, we used a guinea pig model of chronic TB infection in which the lung lesions were similar to those in human TB disease. In the guinea pig TB model, a daily dose of 100 mg delamanid/kg of body weight for 4 or 8 weeks demonstrated strong bactericidal activity against Mycobacterium tuberculosis Importantly, histological examination revealed that delamanid killed TB bacilli within hypoxic lesions of the lung. The combination regimens containing delamanid with rifampin and pyrazinamide or delamanid with levofloxacin, ethionamide, pyrazinamide, and amikacin were more effective than the standard regimen (rifampin, isoniazid, and pyrazinamide). Our data show that delamanid is effective in killing both growing and dormant bacilli in vitro and in the guinea pig TB model. Adding delamanid to current TB regimens may improve treatment outcomes, as demonstrated in recent clinical trials with pulmonary multidrug-resistant (MDR) TB patients. Delamanid may be an important drug for consideration in the construction of new regimens to shorten TB treatment duration.

ТУБЕРКУЛЕЗ У ЖЕНЩИН, НАХОДЯЩИХСЯ В ИСПРАВИТЕЛЬНЫХ УЧРЕЖДЕНИЯХ УГОЛОВНО-ИСПОЛНИТЕЛЬНОЙ СИСТЕМЫ РОССИЙСКОЙ ФЕДЕРАЦИИ

Epidemiological indicators for tuberculosis in men and women over 10 years were analyzed. The incidence of TB, TB prevalence and mortality of female TB-patients in prison is lower than that of men, but do not have the tendency to decrease. Among women, in contrast to men, increases the proportion of patients with chronic TB. The proportion of new cases of destructive forms of TB among women is lower than among men. Lethality of patients with TB decreased among both men and women. Among female TB-patients a higher proportion of patients with co-infection TB and HIV are registered.

TIM3 Mediates T Cell Exhaustion during Mycobacterium tuberculosis Infection.

While T cell immunity initially limits Mycobacterium tuberculosis infection, why T cell immunity fails to sterilize the infection and allows recrudescence is not clear. One hypothesis is that T cell exhaustion impairs immunity and is detrimental to the outcome of M. tuberculosis infection. Here we provide functional evidence for the development T cell exhaustion during chronic TB. Second, we evaluate the role of the inhibitory receptor T cell immunoglobulin and mucin domain–containing-3 (TIM3) during chronic M. tuberculosis infection. We find that TIM3 expressing T cells accumulate during chronic infection, co-express other inhibitory receptors including PD1, produce less IL-2 and TNF but more IL-10, and are functionally exhausted. Finally, we show that TIM3 blockade restores T cell function and improves bacterial control, particularly in chronically infected susceptible mice. These data show that T cell immunity is suboptimal during chronic M. tuberculosis infection due to T cell exhaustion. Moreover, in chronically infected mice, treatment with anti-TIM3 mAb is an effective therapeutic strategy against tuberculosis.

Histomorphological spectrum of paediatric surgical lesions: experience at a tertiary care centre

Aim: The aim of this study is to understand the profile of paediatric surgical pathology specimens in our region. There is paucity of studies on the histologic review in India. Materials & Methods: This is a retrospective analysis of 462 cases of paediatric surgical specimens (Age group 0-12 years) in surgical pathology in a tertiary care centre encountered over a period of one and a half years. Results: Out of 462 cases, 353(76.41%) cases belonged to acquired conditions (both Non-neoplastic and Neoplastic), and 109 (23.59%) cases belonged to congenital and developmental conditions. Males constituted 62.55% while females 37.45%. Gastrointestinal tract was most commonly affected (40.26%) followed by head & neck region (26.19%) and renal & urogenital system (15.58% each). Among acquired conditions (Non-neoplastic and Neoplastic), the most common diagnosed condition was Appendicitis (27.19%) followed by Chronic tonsillitis (24.93%), Inguinal hernia (6.79%), Hydrocele (4.25%), Hemangioma (3.68%), Nonspecific lymphadenitis (3.39%), Empyema (2.83%), TB lymphadenitis and Juvenile polyp (2.55% each) etc. Among acquired neoplastic lesions, there were 4 cases of Wilms tumor, 4 cases of Osteochondroma, 1 case of Fibrous dysplasia, 2 cases of NHL, 1 case of Retinoblastoma, 6 cases of Lipoma and 1 case of Infantile rhabdomyosarcoma. Among congenital and developmental conditions, the most common condition was Hirschsprung’s disease (18.35%), Pelvic ureteric junction obstruction (11%), Meckel’s diverticulum & anorectal malformations (9.17% each), Testicular anomalies & enterogenous cyst (6.42% each) etc. Conclusion: The histopathological patterns of surgical pathology specimens in childhood differ from adults. There are very few published reports on histopathological profile of paediatric surgical pathology specimens in India. Hence, we conducted this study to characterize the histopathological types of surgical pathology specimens among the paediatric patients. Key

Tuberculosis situation in Ismailia governorate (2002–2012) before and after Direct Observed Therapy Short Course Strategy (DOTS)

BackgroundTuberculosis (TB) is a major cause of illness and death worldwide, especially in Asia and Africa. In the early 1990s tuberculosis control in Egypt faced many problems. Major progress in global tuberculosis control followed the widespread implementation of the DOTS strategy. AimThe objective of this work was to study the tuberculosis situation in the Ismailia governorate from 2002 to 2012 before and after Direct Observed Therapy Short Course Strategy (DOTS). MethodsThis was a retrospective clinical cohort study carried out at the Ismailia governorate. The registered data about all TB cases over a period of 10years (2002–2012) before and after the application of DOTS were collected from the chest hospital and TB registration units.Percentages of cure treatment significantly increased after DOTS (55.3%) than before (40.5%) (P<0.01). On the other hand, complete, failure, death, default and transfer out decreased after DOTS (32.5%, 1.3%, 6.3%, 2.5% and 2.0% respectively) than before it (38.7%, 3.1%, 6.9%, 7.2% and 4.0% respectively), the results were not significant (P>0.05) for all of them. The mean values of incidence rates (new and relapse cases, all cases and new smear positive pulmonary TB cases) of TB highly significantly (P<0.01) decreased after the application of DOTS for all of them except new smear positive pulmonary TB cases (P>0.05). Also, the cure rate and treatment success rate significantly increased (P<0.05), while retreatment TB cases rate, default rate, transfer out rate and retreatment failure rate did not significantly decreased (P>0.05) for all of them except retreatment failure rate (chronic TB rate) (P<0.05). Finally new pulmonary TB cases with no smear result significantly (P<0.05) decreased after DOTS. ConclusionThe introduction of DOTS in the Ismailia governorate has led to a significant increase in the treatment success (88.07%) which is higher than the WHO target (85%), and a decrease in the default and failure rates.

Investigation of Immune Biomarkers Using Subcutaneous Model of M. tuberculosis Infection in BALB/c Mice: A Preliminary Report.

Evaluation and screening of vaccines against tuberculosis depends on development of proper cost effective disease models along with identification of different immune markers that can be used as surrogate endpoints of protection in preclinical and clinical studies. The objective of the present study was therefore evaluation of subcutaneous model of M.tuberculosis infection along with investigation of different immune biomarkers of tuberculosis infection in BALB/c mice. Groups of mice were infected subcutaneously with two different doses : high (2×106 CFU) and low doses (2×102 CFU) of M.tuberculosis and immune markers including humoral and cellular markers were evaluated 30 days post M.tuberculosis infections. Based on results, we found that high dose of subcutaneous infection produced chronic disease with significant (p<0.001) production of immune markers of infection like IFNγ, heat shock antigens (65, 71) and antibody titres against panel of M.tuberculosis antigens (ESAT-6, CFP-10, Ag85B, 45kDa, GroES, Hsp-16) all of which correlated with high bacterial burden in lungs and spleen. To conclude high dose of subcutaneous infection produces chronic TB infection in mice and can be used as convenient alternative to aerosol models in resource limited settings. Moreover assessment of immune markers namely mycobacterial antigens and antibodies can provide us valuable insights on modulation of immune response post infection. However further investigations along with optimization of study protocols are needed to justify the outcome of present study and establish such markers as surrogate endpoints of vaccine protection in preclinical and clinical studies in future.

Rapid rebound of the Treg compartment in DEREG mice limits the impact of Treg depletion on mycobacterial burden, but prevents autoimmunity.

The development of an effective vaccine against tuberculosis (Tb) represents one of the major medical challenges of this century. Mycobacterium bovis Bacille Calmette-Guerin (BCG), the only vaccine available at present, is mostly effective at preventing disseminated Tb in children, but shows variable protection against pulmonary Tb, the most common form in adults. The reasons for this poor efficacy are not completely understood, but there is evidence that T regulatory cells (Tregs) might be involved. Similarly, Tregs have been associated with the immunosuppression observed in patients infected with Tb and are therefore believed to play a role in pathogen persistence. Thus, Treg depletion has been postulated as a novel strategy to potentiate M. bovis BCG vaccination on one side, while on the other, employed as a therapeutic approach during chronic Tb infection. Yet since Tregs are critically involved in controlling autoimmune inflammation, elimination of Tregs may therefore also incur the danger of an excessive inflammatory immune response. Thus, understanding the dynamics and function of Tregs during mycobacterial infection is crucial to evaluate the potential of Treg depletion as a medical option. To address this, we depleted Tregs after infection with M. bovis BCG or Mycobacterium tuberculosis (Mtb) using DEREG mice, which express the diphtheria toxin (DT) receptor under the control of the FoxP3 locus, thereby allowing the selective depletion of FoxP3+ Tregs. Our results show that after depletion, the Treg niche is rapidly refilled by a population of DT-insensitive Tregs (diTregs) and bacterial load remains unchanged. On the contrary, impaired rebound of Tregs in DEREG × FoxP3GFP mice improves pathogen burden, but is accompanied by detrimental autoimmune inflammation. Therefore, our study provides the proof-of-principle that, although a high degree of Treg depletion may contribute to the control of mycobacterial infection, it carries the risk of autoimmunity.

Lead Optimization of 1,4-Azaindoles as Antimycobacterial Agents

In a previous report, we described the discovery of 1,4-azaindoles, a chemical series with excellent in vitro and in vivo antimycobacterial potency through noncovalent inhibition of decaprenylphosphoryl-β-d-ribose-2'-epimerase (DprE1). Nevertheless, high mouse metabolic turnover and phosphodiesterase 6 (PDE6) off-target activity limited its advancement. Herein, we report lead optimization of this series, culminating in potent, metabolically stable compounds that have a robust pharmacokinetic profile without any PDE6 liability. Furthermore, we demonstrate efficacy for 1,4-azaindoles in a rat chronic TB infection model. We believe that compounds from the 1,4-azaindole series are suitable for in vivo combination and safety studies.

Evolução da pneumonia lipoide exógena em crianças: aspectos clínicos e radiológicos e o papel da lavagem broncoalveolar

To present aspects of the evolution of lipoid pneumonia in children, based on clinical, radiological and bronchoalveolar lavage fluid findings, emphasizing the importance of bronchoalveolar lavage for the diagnosis and treatment. We included 28 children, with a mean age of 20 months (range, 1-108 months), diagnosed with chronic pneumonia refractory to antimicrobial therapy, with TB or with a combination of the two. Most of the children had at least one risk factor for aspiration, and all of them had a history of mineral oil ingestion for intestinal constipation (23/28) or complicated ascaridiasis (5/28). Clinical evaluations, tomographic evaluations and analyses of bronchoalveolar lavage fluid were carried out at the beginning of treatment and throughout a follow-up period of 24 months. Tachypnea and cough were the most common symptoms. The most common radiological alterations were areas of consolidation (23/28), perihilar infiltrates (13/28) and hyperinflation (11/28). Chest CT scans showed areas of consolidation with air bronchogram (24/28), decreased attenuation in the areas of consolidation (16/28), ground-glass opacities (3/28) and crazy-paving pattern (1/28). In the analysis of the bronchoalveolar lavage fluid, Sudan staining revealed foamy macrophages, confirming the diagnosis of lipoid pneumonia. After treatment with multiple bronchoalveolar lavages (mean = 9.6), 20 children became asymptomatic, 18 of those presenting normal tomographic images. A diagnosis of lipoid pneumonia should be considered in patients with chronic refractory pneumonia or TB, especially if there is a history of mineral oil ingestion. Bronchoscopy with multiple bronchoalveolar lavages was an efficient treatment for the clearance of mineral oil from the lung parenchyma and the prevention of fibrosis. This strategy contributed to reducing the morbidity of lipoid pneumonia, which remains a rare diagnosis.

Molecular detection of rifampin and isoniazid resistance to guide chronic TB patient management in Burkina Faso

BackgroundDrug-resistant tuberculosis (DR-TB) is considered a real threat to the achievement of TB control. Testing of mycobacterial culture and testing of drug susceptibility (DST) capacity are limited in resource-poor countries, therefore inadequate treatment may occur, favouring resistance development. We evaluated the molecular assay GenoType® MTBDRplus (Hain Lifescience, Germany) in order to detect DR-TB directly in clinical specimens as a means of providing a more accurate management of chronic TB patients in Burkina Faso, a country with a high TB-HIV co-infection prevalence.MethodsSamples were collected in Burkina Faso where culture and DST are not currently available, and where chronic cases are therefore classified and treated based on clinical evaluation and sputum-smear microscopy results. One hundred and eight chronic TB patients (sputum smear-positive, after completing a re-treatment regimen for pulmonary TB under directly observed therapy) were enrolled in the study from December 2006 to October 2008. Two early morning sputum samples were collected from each patient, immediately frozen, and shipped to Italy in dry ice. Samples were decontaminated, processed for smear microscopy and DNA extraction. Culture was attempted on MGIT960 (Becton Dickinson, Cockeysville, USA) and decontaminated specimens were analyzed for the presence of mutations conferring resistance to rifampin and isoniazid by the molecular assay GenoType® MTBDRplus.ResultsWe obtained a valid molecular test result in 60/61 smear-positive and 47/47 smear-negative patients.Among 108 chronic TB cases we identified patients who (i) harboured rifampin- and isoniazid-susceptible strains (n 24), (ii) were negative for MTB complex DNA (n 24), and (iii) had non-tuberculous mycobacteria infections (n 13). The most represented mutation conferring rifampin-resistance was the D516V substitution in the hotspot region of the rpoB gene (43.8% of cases). Other mutations recognized were the H526D (15.6%), the H526Y (15.6%), and the S531L (9.4%).All isoniazid-resistant cases (n 36) identified by the molecular assay were carrying a S315T substitution in the katG gene. In 41.7% of cases, a mutation affecting the promoter region of the inhA gene was also detected.ConclusionThe GenoType® MTBDRplus assay performed directly on sputum specimens improves the management of chronic TB cases allowing more appropriate anti-TB regimens.

HIV-1 compromises CD8+T cell effector function in Mtb- infected lung (129.26)

Abstract Tuberculosis (TB) remains a global health problem with more than 8 million new cases and 1 million deaths per year worldwide. HIV-1 infected persons have a greatly increased risk of Mycobacterium tuberculosis (M. tb) co-infection. A key feature of M.tb infection is the formation of granulomas, cellular accumulations composed of macrophages, epithelial cells, and a surrounding mantle of T cells, which are important for the containment of infection. The cytolytic T cell effector molecules, perforin and granulysin, play a critical role in protective immunity to M.tb. It has been shown that decreased expression of perforin and granulysin at the site of pulmonary disease is linked to chronic TB. However, the effects of HIV-1 on CD8+T cells numbers and expression of cytolytic effector molecules such as perforin and granulysin in M. tb induced granulomas has not been explored. In this study, we used immunohistological techniques to assess the differences in T cell populations, granulysin expression, and pathology in tissue sections from Mtb- and Mtb/HIV-1-infected human lung. Our results indicate that granulomas from persons with TB/HIV-1 are characterized by dysregulated T cell organization and poor expression of granulysin. We propose that defective granulysin expression by M.tb-specific CD8+T cells contributes to the development of disease in TB/HIV-1 co-infected persons.