Peroxynitrite in Sarcoidosis: Relation to Mycobacterium Stationary Phase.

Sarcoidosis (SA) is a disorder of unknown aetiology. Mycobacterium tuberculosis heat shock proteins (Mtb-hsp) have been considered as causative agents of SA. The role of Mtb-hsp in the immune response in SA has not been investigated. Mtb-hsp-stimulated T-cell subsets and Th1/Th2 cytokine patterns in the supernatant from peripheral blood mononuclear cell cultures from 22 SA patients, 20 tuberculosis (TB) patients and 20 healthy volunteers were compared using flow cytometry. In unstimulated cultures, a significantly higher percentage of CD8(+)alphabeta(+)T-cells were present in SA versus controls. Similarly there was a significantly increased IL-6 and decreased IL-4 level in SA and significantly lower INF-gamma, IL-2, IL-4, IL-10 production in TB versus controls. After Mtb-hsp stimulation, there was a significantly increased TNF-alpha, IL-6, IL-10 and decreased INF-gamma, IL-2, IL-4 production in SA and significantly increased TNF-alpha, IL-6 concentrations in TB versus controls. CD8(+)gammadelta(+)IL-4(+)T-cells were detected significantly less often in Mtb-hsp-induced cultures in SA versus controls. Comparing SA versus TB, CD4(+)gammadelta(+)TCR-cells were significantly increased in Mtb-hsp-induced cultures in TB versus controls and SA. Before stimulation, significantly increased IL-6, IL-10 and decreased IL-4 level in SA versus TB was revealed, whereas Mtb-hsp stimulation caused significantly increased IL-10 and decreased IL-4 concentrations in SA. After Mtb-hsp stimulation, increased levels of pro-inflammatory cytokines, TNF-alpha and IL-6 were found in sera from SA and TB patients in comparison with healthy controls; SA patients demonstrated the lowest levels of IL-4 and the highest levels of IL-10.

Mycobacterium tuberculosis Infections in Solid Organ Transplantation

Changed phagocytic activity and pattern of Fcγ and complement receptors on blood monocytes in sarcoidosis

Background: Tuberculosis (TB) and sarcoidosis are two chronic, systemic, and granulomatous diseases that have similar pulmonary and extrapulmonary manifestations. Given the similarities between the two diseases, it is suggested that the Mycobacterium genome may play a role in sarcoidosis. Identification of Mycobacterium species isolated from sarcoidosis patients optimizes the treatment process of patients. The present study investigates biopsy tissue samples from patients with sarcoidosis for the presence of Mycobacterium in the tissues. Methods: This descriptive cross-sectional study was performed on all sarcoidosis patients referred to Masih Daneshvari Hospital during 2017–2018 (401 patients). All patients who were outpatients or hospitalized in the bronchoscopy ward of Masih Daneshvari Hospital underwent transbronchial needle aspiration (TBNA). The cause of the TBNA procedure in patients was the presence of bilateral hilar lymphadenopathy in patients. Out of 401 patients with sarcoidosis, five patients showed positive smear or culture for Mycobacterium and were excluded from the study. Finally, 396 patients were included in the study. Results: The majority of participants in the present study were female (56.1%) with a mean age of 48.2 years and a mean ACE index of 73.4. Hypertension was found in16.2% of patients with sarcoidosis. 16.4% of patients had hyperlipidemia and 4.3% of patients had a history of ischemic heart disease. Redness of the eyes was seen in 13.1% of patients and 12.4% of the patients had diabetes. Also, 43.4% of the patients had skin lesions. Out of a total of 396 patients with sarcoidosis in the present study, 3.5% of the lymph and lung tissues of sarcoidosis patients (who were not infected with Mycobacterium tuberculosis by clinical signs and culture) contained amplifiable MTB DNA. Conclusion: In the present study and many previous related studies, the Mycobacterium genome has been observed in lymph node samples of mediastinal nodes and lung tissue of sarcoidosis patients. One of the strengths of the present study, which can make the results very reliable, is the high sample size in this study. It is suggested that similar studies with the use of control groups in large samples, investigate the presence of the Mycobacterium genome and its type in sarcoidosis patients.

Studies on the role of mannose-binding lectin 2 (MBL2) in individuals infected with tuberculosis (TB) remain limited. This study aimed to compare MBL2 gene expression and protein concentration between active and latent TB cases and to assess the influence of sex on these differences. This cross-sectional study involved 39 newly diagnosed active pulmonary TB patients and 25 individuals with latent TB who were household contacts. MBL2 gene expression was evaluated using a relative quantitative polymerase chain reaction method. MBL protein levels were measured using the enzyme-linked immunosorbent assay. Among female participants, MBL2 gene expression was significantly lower in those with active TB compared to those with latent TB (P = 0.02). In male participants, no significant difference was observed (P = 0.333). Similarly, MBL protein levels tended to be lower in females with active TB than in those with latent TB, though this difference was not statistically significant (median [range]: 124.78 [65.62-499.79] vs. 208.49 [99.85-498.65] ng/mL, P = 0.099). In males, no significant difference in MBL protein levels was detected between the active TB and latent TB groups (206.86 [59.11-526.77] vs. 143.55 [65.85-290.7] ng/mL, P = 0.285). This study highlights the influence of sex on the expression of the MBL2 gene and plasma protein levels in TB patients. A lower expression of the MBL2 gene in active TB cases compared to latent TB cases was observed exclusively in women.

<b>Background:</b> Understanding the CD8+ response against Mycobacterium tuberculosis (MTB) may be a key to improved TB diagnostics and vaccine development. <b>Aims and Objectives:</b> To detect a CD8+ T-cell response against Mycobacterium tuberculosis (MTB) in active tuberculosis (TB) and latent TB (LTBI), in HLA A*02 positive patients. <b>Methods:</b> We identified possible epitopes (antigen fragments) from 9 MTB antigens (Ag85B, ESAT-6, EsxH, Hsp65, EsxJ, rv1490, rv1614, rv2626c and 16 kDa antigen) by searching the literature, and by computer prediction of likely binding to MHC-1. We selected 24 epitope candidates, from which we constructed MHC multimers (Dextramers). Peripheral blood mononuclear cells (PBMC) from 7 TB-patients, 16 LTBI patients and 8 MTB-exposed, IGRA-negative, healthy subjects (HE), all HLA A*02 positive, were stained with the Dextramers and with anti-CD8 and anti-CD3, and analyzed on a flow cytometer. The MTB epitopes were analyzed in 5 pools (3-7 epitopes each). Positive responses included &gt;0.001 % of CD8+, CD3+ cells, supported by inspection of flow cytometry plots. <b>Results:</b> MTB-specific CD8+ T-cells were detected more often in TB patients (57%) than in LTBI patients (41%) and in HE (25%), though differences were insignificant. CD8+ responses were seen against a broad spectrum of antigens, including not previously described Rv2626c epitopes. Reactivity against multiple antigens was observed in 3 of 7 LTBI patients and in 2 of 4 TB patients with detectable CD8+ responses. <b>Conclusion:</b> Broad diversity in epitope-specificity was seen among MTB-specific CD8+ T-cells in HLA A*02, consistent with previous reports. A trend of more common detection of MTB-specific CD8+ cells in TB than in LTBI and HE was found.

Tuberculosis (TB) remains one of the most severe infectious diseases. It is still of paramount importance to establish more accurate, rapid, and efficient diagnostic methods. Since infection with Mycobacterium tuberculosis (M. tb) is largely mediated through the respiratory tract, IgA responses against mycobacterial proteins are worthy of investigation for their potential clinical utility. In this study, the IgA response targeting lipoprotein Z (LppZ) was determined by using a homemade ELISA with plasma of TB patients (N = 125), LTBI individuals (N = 92), healthy controls (HCs) (N = 165), as well as TB patients undergoing anti-TB treatment (N = 9). In parallel the antigen-specific IFN-γ release from PBMCs triggered by LppZ and M. tb-specific ESAT-6 or CFP-10 was detected by using an ELISPOT assay. It was found that the LppZ-specific IgA level was dramatically higher in TB patients than in HCs (p < 0.0001). Compared to that before anti-TB treatment, the LppZ-specific IgA level decreased substantially after 2 months of anti-TB treatment (p = 0.0297) and remained at low levels until the end of the treatment. What is more, pulmonary TB patients exhibited significantly higher LppZ-specific IgA-values than extra-pulmonary TB patients (p = 0.0296). Interestingly, the LppZ-specific IgA-values were negatively correlated to the amounts of IFN-γ released in response to LppZ with statistical significance (r = −0.5806, p = 0.0002). LppZ-specific IgA level was also higher in LTBI individuals than in HCs (p < 0.0001). Additionally there were some PPD+ HC individuals with high LppZ-specific IgA levels but the potential of this assay for identifying leaky LTBI in PPD+ HCs needs to be further investigated through follow-up studies. The sensitivity of detecting TB solely with ESAT-6 or CFP-10-specific IFN-γ release was increased by including the LppZ-specific IgA results, respectively, from 86.11 to 100% and 88.89 to 100%; the sensitivity of screening for LTBI was increased from 80.36 to 83.93% and 57.14 to 69.64%, respectively. The higher LppZ-specific IgA responses in TB and LTBI populations than in controls indicated high immunoreactivity to LppZ upon M. tb infection. Although the assay was not efficient enough for independent application in sero-diagnosis, LppZ-specific IgA might become a complementary biomarker for the improvement of TB and LTBI screening.

BackgroundTo describe the trends of Type 2 Diabetes with Pulmonary Tuberculosis (T2DM-TB) patients from 2013 to 2022 and to investigate the impact of COVID-19 lockdown on glycemic control and associated factors in T2DM-TB. MethodsIn this population-based study of the First Affiliated Yijishan Hospital of Wannan Medical College in China, we described the 10-year trends of patients diagnosed with T2DM-TB. We included patients diagnosed with TB, T2DM-TB and T2DM-TB patients for comparative analysis, aged 15 years or older. Data were missing, and both multidrug-resistant (MDR) TB patients and non-T2DM patients were excluded from our study. ResultsWe pooled Type 2 Diabetes (T2DM) and Tuberculosis (TB) data from The First Affiliated Yijishan Hospital of Wannan Medical College in China, gathered between January 1, 2013, and December 31, 2022. The data included 14,227 T2DM patients, 6130 TB patients, and 982 T2DM-TB patients. During the past 10 years, the number of inpatients with TB decreased, while the number of patients with T2DM and T2DM-TB increased year by year. To rule out any influence factors, we analyzed the ratio of the three groups. The ratio of TB/T2DM decreased year by year (p < 0.05), while the ratio of TB-T2DM/TB increasing year by year (p = 0.008). During the COVID-19 epidemic period, there was no significant change in the ratio of TB-T2DM/T2DM (p = 0.156). There was no significant change in the proportion of male patients with TB and TB-T2DM (p = 0.325; p = 0.190), but the proportion of male patients with T2DM showed an increasing trend (p < 0.001). The average age of TB patients over the past 10 years was 54.5 ± 18.4 years and showed an increasing trend year by year (p < 0.001). However, there was no significant change in the age of T2DM or TB-T2DM patients (p = 0.064; p = 0.241). Patients data for the first (2013–2017) and the last (2018–2022) five years were compared. We found that the number of T2DM and TB-T2DM in the last five years was significantly higher than in the first five years, but the number of TB was significantly lower than in the first five years. There is a significant statistical difference in the proportion of TB/T2DM and TB-T2DM/TB, which is similar to the previous results. The average age (56.0 ± 17.6 years) of TB patients in the last five years is significantly higher than in the first five years (53.1 ± 18.9) (p < 0.001). The number of male patients with T2DM in the last five years is higher than that in the first five years, with significant difference (p < 0.001). ConclusionThe trends of T2DM-TB among hospitalized TB patients have increased significantly over the past 10 years, which may be related to the increase in the number of T2DM cases. The COVID-19 pandemic has been effective in controlling the transmission of TB, but it has been detrimental to the control of T2DM. Male patients with T2DM and elderly TB patients are the key populations for future prevention and control efforts.

IntroductionThe rising prevalence of Diabetes mellitus (DM) in high TB‐endemic countries may adversely affect sustainability of TB control since DM constitutes a risk factor for development of active tuberculosis (TB). The impact of DM on TB specific adaptive immune responses remains poorly addressed, particularly in people living in Sub‐Saharan countries. We performed a functional characterization of TB specific cellular immune response in Tanzanian subjects with active or latent Mycobacterium tuberculosis (Mtb) infection stratified by their diabetic status.MethodsHIV negative active TB patients (≥18 years) with Xpert MTB/RIF positive pulmonary TB were included before starting TB treatment in Dar es Salaam, Tanzania between April and December 2013. HIV negative healthy controls latently infected with TB but without past TB history were also included. Active and latent TB patients were stratified in two groups according to their diabetic status. Peripheral Blood Mononuclear cells were stimulated with either live M. bovis BCG or Mtb‐specific peptide pools and analyzed by intracellular cytokine staining and polychromatic flow cytometry.ResultsOur results show a lower frequency of IFN‐γ CD4+ T cells in patients with active TB and DM compared to patients with active TB only after live M. bovis BCG (p = 0.04) but not after Mtb peptide pools re‐stimulation. Irrespective of TB status, level of glycaemia is selectively inversely correlated with IFN‐γ and TNF‐α CD4+ T cell production (p = 0.02 and p = 0.03) after live M. bovis BCG stimulation.ConclusionsThese results support the hypothesis that hyperglycaemia negatively impacts antigen processing and/or presentation of whole mycobacteria delaying secretion of key cytokines involved in TB immunity.

Sarcoidosis (SA) is a granulomatous disorder of an unknown etiology. Mycobacterium tuberculosis heat shock proteins (Mtb-hsp), considered as causative agents, play an important role in apoptosis. A role for apoptosis has been proposed in pathogenesis of SA and tuberculosis (TB) granuloma formation but results remain controversial. Differences in Mtb-hsp-induced apoptosis between SA, TB, and healthy subjects found in this study might put some light on the etiology of SA. Early apoptotic peripheral blood mononuclear cells (PBMC) were determined in 22 SA patients, 20 TB patients, and 20 healthy volunteers by flow cytometry (Annexin-V-FITC). Our results revealed that spontaneous apoptosis of monocytes and CD8+ T-cells was comparable between tested groups. Apoptosis of unstimulated CD4+ T-cells was significantly lower in TB versus controls and insignificantly lower versus SA. Mtb-hsp- and PHA (Phytohemagglutinin)-induced monocytes apoptosis was significantly lower in TB versus controls and SA. Mtb-hsp-induced CD4+ T-cell apoptosis was significantly lower in TB versus controls and SA. There were no differences of PHA-induced CD4+ T-cell and CD8+ T-cell apoptosis between tested groups. Apoptosis of Mtb-hsp-induced CD8+ T-cells was significantly lower in TB and SA versus controls. Analysis of PBMC apoptosis before and after stimulation in each tested group revealed that, in contrast to TB, sarcoid monocytes were resistant to Mtb-hsp- and PHA-induced apoptosis and CD4+ T-cells were resistant to PHA- but not Mtb-hsp-induced apoptosis. CD8+ T-cell apoptosis, before and after Mtb-hsp or PHA stimulation, was significantly increased in all tested groups. It seems likely that dysregulated apoptosis of CD4+ T-cells and resistant apoptosis monocytes may be involved in pathogenesis of SA.

Differentiating between active and latent tuberculosis with chest computed tomography

OneTouch® Comfort™ : un nouveau système de prélèvement capillaire

<b>Objectives:</b> To investigate the immune response against <i>M. tuberculosis (MTB)</i> by Interferon Gamma Release Assays (IGRAs) in Tuberculosis (TB) and Latent TB Infected (LTBI) patients exposed to cigarette smoke. <b>Methods:</b> A cross sectional study from two clinics in Barcelona. Subjects included were: (i) 216 active pulmonary TB, (ii) 309 asymptomatic contact tracing individuals. At first visit blood was collected for IGRAs. The impact of cigarette smoke on MTB immune response was analyzed as the median of IFN-γ secreted (UI/mL) in QFN-G-IT and as the number (median) of T-cells producing IFN-γ [spot forming cells (SFC)] in T-SPOT.TB. <b>Results:</b> False-negative results in TB patients were higher in smokers (aOR:4.42; p&lt;0.0001). Smoker contacts had higher LTBI prevalence (aOR:11.98; p&lt;0.001). In non smoker TB patients the immune response [QFN.GIT: 3.37 (1.4-9.5); T-SPOT.TB: 75.5 (30.0-134.0)] was higher (p&lt;0.0001) than in smokers [(QFN.GIT: 0.93 (0.2-3.3); T-SPOT.TB: 38.5 (13.0-99.0)]. In LTBI contacts the IFN-γ response was higher (p&lt;0.05) in non smokers [QFN.GIT: 4.6 (0.8-10.6);T-SPOT.TB: 50.0 (25.0-144.0)] than in smokers [QFN.GIT: 1.3 (0.5-4.0; T-SPOT.TB: 31.0 (17.0-53.0)]. A negative dose-dependent relation was observed between the smoking quantity and the IFN-γ response analyzed by QFN-G-IT and T-SPOT.TB for active TB patients and contacts with LTBI. <b>Conclusions:</b> Tobacco exposure was associated with IGRAs false-negative results in TB patients and was an independent risk factor for LTBI acquisition. A less IFN-γ response was observed in smokers than non-smokers. An increment on tobacco smoking produces a reduction on immune responses observed by both IGRAs. PII –Tabaquismo. 2013 SEPAR grant.

Objectives:This study was planned to define the clinical profile of pulmonary tuberculosis (PTB) patients having renal amyloidosis, to identify the factors responsible for development of amyloidosis, to detect the time period between onset of amyloidosis and PTB, and analyze clinical features of amyloidosis in PTB patients for early diagnosis and timely assessment.Materials and Methods:Patients of PTB having pedal edema, proteinuria, and grossly diseased kidneys on ultrasound abdomen were subjected to renal biopsy and appropriate biochemical investigations. Clinical profile of biopsy proven amyloidosis cases was analyzed.Results:There were 43 patients (32 males, 11 females, age range 20–65 years) having PTB with pedal edema, proteinuria, and renal medical disease on abdominal ultrasound where amyloidosis was confirmed by renal biopsy. The total duration of illness ranged from two months to seven years (mean 2.25 years) and was less than five years in 93% patients. All patients had significant proteinuria. Nephrotic syndrome was seen in 23, hypertension in 19, hypoalbuminemia in 33, hypercholesterolemia in 29, and deranged renal functions in 32 patients. Ninety percent patients had moderate to far advanced pulmonary lesions on chest radiography with smear positivity in 21 patients.Conclusions:Renal amyloidosis is an important complication of PTB and should be suspected clinically in patients presenting with a triad of pedal edema, proteinuria, and medical renal disease on ultrasound. Contrary to general belief, renal amyloidosis may occur in PTB patients having disease for relatively shorter duration, and even if adequately treated.

Introduction: RNTCP estimates MDR in India as 2-3% in new and 12-17% in treated cases. Mumbai is a hot spot for TB drug resistance in India. A prior study evaluating MDR patients revealed 29.4% MDR, 56.8% pre XDR, 10.6% XDR and 5.6% XXDR in Mumbai. Aims & objectives: To evaluate the patterns of drug resistance in TB patients following up at our hospital. Methods: A retrospective observational study of drug susceptibility testing (DST) results on positive TB MGIT cultures of 53 TB patients from January 2015 to December 2015) was conducted. Patients were classified into 5 categories based on the DST results as drug sensitive TB, MDR TB, pre XDR TB, XDR TB & XXDR TB. Results: 43 patients (81.13%) had pulmonary and 10 patients (18.87%) had extra-pulmonary TB. Of the 43 pulmonary cases, 18(41.86%) had drug sensitive TB and 25(58.13%) had drug resistant TB. In drug resistant pulmonary cases, pre XDR TB was found in 15(60%), MDR TB in 6(24%), XXDR TB in 2(8%), XDR TB in 1(4%) and INH mono resistance in 1(4%). Amongst 10 extra-pulmonary TB cases, 4(40%) had drug sensitive TB and 6(60%) had drug resistant TB of which pre XDR TB was found in 4(66.66%), XXDR in 1(16.66%) and INH mono resistance in 1(16.66%). Overall incidence of pre-XDR TB was 61.9% (19 Patients). Conclusion: Incidence of pre XDR TB is on the rise among drug resistant pulmonary and extra-pulmonary TB patients in Mumbai. This necessitates the need for complete drug susceptibility testing and not relying solely on molecular techniques that will identify MDR only. Individualized drug regimens for these patients based on DST will help in curtailing transmission of resistant strains within the community.