Efficacy Of Tuberculosis Treatment Research Articles

TB/FLU-06E Influenza Vector-Based Vaccine in the Complex Therapy of Drug-Susceptible and Drug-Resistant Experimental Tuberculosis.

The steady rise of drug-resistant tuberculosis (TB), which renders standard therapy regimens ineffective, necessitates the development of innovative treatment approaches. Immunotherapeutic vaccines have the potential to effectively regulate the anti-TB immune response and enhance the efficacy of anti-TB treatment. In the present study, we aimed to evaluate the potency of the mucosal vector vaccine TB/FLU-06E as part of a complex treatment regimen for drug-susceptible (DS) or drug-resistant (DR) tuberculosis in C57BL/6 mice. Incorporating TB/FLU-06E into the treatment protocol significantly increased the effectiveness of therapy for both forms of tuberculosis. It was evidenced by higher survival rates and reduced pulmonary bacterial load (1.83 lg CFU for DS tuberculosis and 0.93 lg CFU for DR tuberculosis). Furthermore, the treatment reduced pathomorphological lesions in the lungs and stimulated the local and systemic T-helper 1 (Th1) and cytotoxic T-lymphocyte (CTL) anti-TB immune responses. Thus, therapeutic immunization with the TB/FLU-06E vaccine significantly enhances the efficacy of tuberculosis treatment, which is particularly important in DR tuberculosis.

Assessment of Comorbidity in Patients with Drug-Resistant Tuberculosis.

A wide range of comorbidities, especially in multidrug-resistant tuberculosis (MDR-TB) and extensively drug-resistant tuberculosis (XDR-TB) patients, markedly complicates selecting effective treatment of tuberculosis (TB) and preventing the development of adverse events. At present, it is impossible to assess the severity of comorbid pathologies and develop indications for the administration of accompanying therapy in TB patients. The aim of this study was to identify the difference in the range of comorbidities between patients with MDR-TB and XDR-TB and assess the impact of comorbidities on TB treatment. Materials and Methods: A retrospective, prospective study was conducted where 307 patients with MDR-TB and XDR-TB pulmonary tuberculosis aged 18 to 75 years who received eTB treatment from 2016 to 2021 in St. Petersburg hospitals were analyzed. The analysis showed that the comorbidity level in MDR-TB and XDR-TB patients with TB treatment success and treatment failure was comparable with the use of the Charlson Comorbidity Index (CCI). The CCI demonstrated declining data in terms of TB treatment outcome period in both groups. A slight predominance of CCI score (3 to 4 points) in XDR-TB (22.7%) vs. MDR-TB (15.4%) patients was found. In the case of an TB treatment failure, the CCI level in MDR-TB vs. XDR-TB patients was characterized by a significantly higher rate of low magnitude (ranging from 1 to 2 points) in 21.1% vs. 4.5% (p < 0.05), which was higher in XDR-TB patients (ranging from 4 to 5 points, in 10.0% vs. 0, χ2 = 33.7 (p < 0.01)). Chronic viral hepatitis B and C infection, cardiovascular pathology, chronic obstructive pulmonary disease, and chronic alcoholism were found to be significant comorbidity factors that influenced the TB treatment success. Conclusions: It is evident that XDR-TB patients comprise a cohort with the most severe disease course due to comorbidities impacting TB treatment efficacy. The obtained data pointed to the need to determine comorbidity severity in patients with drug-resistant Mbt prior to administering TB treatment schemes.

Optimizing tuberculosis treatment efficacy: Comparing the standard regimen with Moxifloxacin-containing regimens.

Tuberculosis (TB) continues to be one of the deadliest infectious diseases in the world, causing ~1.5 million deaths every year. The World Health Organization initiated an End TB Strategy that aims to reduce TB-related deaths in 2035 by 95%. Recent research goals have focused on discovering more effective and more patient-friendly antibiotic drug regimens to increase patient compliance and decrease emergence of resistant TB. Moxifloxacin is one promising antibiotic that may improve the current standard regimen by shortening treatment time. Clinical trials and in vivo mouse studies suggest that regimens containing moxifloxacin have better bactericidal activity. However, testing every possible combination regimen with moxifloxacin either in vivo or clinically is not feasible due to experimental and clinical limitations. To identify better regimens more systematically, we simulated pharmacokinetics/pharmacodynamics of various regimens (with and without moxifloxacin) to evaluate efficacies, and then compared our predictions to both clinical trials and nonhuman primate studies performed herein. We used GranSim, our well-established hybrid agent-based model that simulates granuloma formation and antibiotic treatment, for this task. In addition, we established a multiple-objective optimization pipeline using GranSim to discover optimized regimens based on treatment objectives of interest, i.e., minimizing total drug dosage and lowering time needed to sterilize granulomas. Our approach can efficiently test many regimens and successfully identify optimal regimens to inform pre-clinical studies or clinical trials and ultimately accelerate the TB regimen discovery process.

The impact of directly observed therapy on the efficacy of Tuberculosis treatment: a Bayesian multilevel approach

Abstract We propose and discuss a Bayesian procedure to estimate causal effects for multilevel observations in the presence of confounding. This work is motivated by an interest in determining the causal impact of directly observed therapy on the successful treatment of Tuberculosis. We focus on propensity score regression and covariate adjustment to balance the treatment allocation. We discuss the need to include latent local-level random effects in the propensity score model to reduce bias in the estimation of causal effects. A simulation study suggests that accounting for the multilevel nature of the data with latent structures in both the outcome and propensity score models has the potential to reduce bias in the estimation of causal effects.

Identifying autophagy-related genes as potential targets for immunotherapy in tuberculosis

Identifying of host-directed targets and molecular markers of immune response for tuberculosis (TB) immunotherapy is urgent and meaningful. Previous studies have demonstrated an important role of autophagy in the course and pathophysiology of TB and is associated with the efficacy of TB treatment. However, its role in TB immunotherapy is still incomplete. The effect of autophagy on intracellular bacteria load was examined in sulforaphane (SFN)-treated THP-1 cells. The immune infiltration was assessed based on public databases. Functional enrichment analysis revealed the pathways involved. LASSO Cox regression analysis was employed to identify hub genes. Moreover, machine learning analysis was used to obtain important targets of TB immunotherapy. Finally, the relationship between hub genes and immune infiltration was assessed, as well as the relevance of chemokines. We found that SFN reduced intracellular bacteria load by enhancing autophagy in THP-1 cells. Thirty-two autophagy-related genes (ARGs) were identified, three types of immune cells (macrophages, neutrophils, and DC cells) were significantly enriched in TB patients, and 6 hub genes (RAB5A, SQSTM1, MYC, MAPK8, MAPK3, and FOXO1) were closely related to TB immune infiltration. The 32 ARGs were mainly involved in autophagy, apoptosis, and tuberculosis pathways. FOXO1, SQSTM1, and RAB5A were identified as important target genes according to the ranking of variable importance, with FOXO1 being a potential autophagy-related target of TB immunotherapy. This study highlights the association between autophagy-related genes and immune infiltration in TB. Three key genes, especially FOXO1, regulated by SFN, will provide new insights into diagnostic and immunotherapy strategies for clinical tuberculosis.

Efficacy of Tuberculosis Treatment in Patients with Drug-Resistant Tuberculosis with the Use of Bedaquiline: The Experience of the Russian Federation

The authors analyzed data published in studies from 2014 to 2022; 41 publications were included in total and 17 articles corresponded to the study design. The results of treatment of 1404 tuberculosis patients with MDR/XDR TB were described. Bedaquiline was used according to the standard scheme with a description of the treatment results after 24-26 weeks. Treatment efficacy was estimated according to accepted criteria. The analysis showed that the treatment efficacy on conversion was achieved in 79.5% of cases (95% Cl 76.5-82.3), and recovery was observed in 82.0% of cases (95% Cl 78.6-85.1). Departure from the therapy was observed in rare cases (9.8%; 95% Cl 7.9-12.2). Deaths were recorded in 6.5% of cases (95% Cl 4.9-8.3), which were associated with the severe disease and concomitant pathology in 74.3%. The development of adverse events was noted in half of the patients (55.7%); however, bedaquiline cancellation occurred in a few cases (7.0%; 95% Cl 3.0-13.0). From analyzing data in patients with MDR and XDR TB, the efficacy of treatment was 89.9% (95% Cl 85.9-93.2) and 71.9% (95% Cl 66.2-77.1), respectively. Use of bedaquiline in treatment makes it possible to achieve recovery of patients with MDR/XDR TB in 82.0% of cases with patients dropping out of treatment in 9.8%. At the same time, in patients with MDR TB, recovery was achieved in 89.9% of cases, while in patients with XDR TB, 71.9% of cases recovered.

Nanoparticle based depletion of Myeloid-derived Suppressor cells in Tuberculosis

Abstract The bacterial pathogen Mycobacterium tuberculosis (Mtb) successfully evades host innate immune responses to cause tuberculosis (TB) disease. Recent studies showed that myeloid-derived suppressor cells (MDSCs) are recruited to the lung after Mtb infection and harbor Mtb. While MDSC-mediated negative regulation of tumor micro-environments in cancer has been extensively studied, in-depth analyses of the phenotype, functions and transcriptional signatures of MDSCs in TB are lacking. In this study we sought to characterize MDSCs in the lungs of Mtb-infected mice and develop tools for depleting MDSCs as a strategy for host-directed therapy. We determined MDSCs in the aerosol mouse model of TB and in human blood using flow cytometry. MDSCs are recruited into the lung as early as 2 weeks post-infection and expresses immunomodulatory mediators associated with immune suppression such as iNOS, IDO and arginase. We recently reported the development of novel synthetic nanoparticle antibodies (SNAbs) that specifically target and deplete MDSCs via antibody-like mechanisms. We used SNAbs to deplete Mtb-induced MDSCs in vitro and in human PBMCs ex vivo. Interestingly, we found that SNAbs efficiently depleted MDSCs in both models. To test the efficacy of SNAb-mediated depletion of MDSCs in mouse lungs in vivo, we first used a murine lung cancer model and found that intra-tracheal delivery of SNAbs successfully depleted MDSCs in vivo. Ongoing studies include evaluating the ability of SNAbs to deplete lung MDSCs following Mtb infection of mice and to assess the impact on lung T cell functions, pathology and Mtb burden. These studies will establish a platform to investigate MDSCs as a target for host-directed therapies that improve TB treatment efficacy. Supported by grant from NIH (1R01AI155023-01A1)

The role of vitamin D deficiency in antituberculous protection

BACKGROUND. The main task of modern phthysiology is a comprehensive search for ways to optimize the etiotropic and the pathogenetic treatment of tuberculosis (TB). The search for improved treatment in addition to etiotropic antimicrobial therapy lies in the plane of improving pathogenetic therapy. Analysis of the available scientific sources suggests that the efficacy of TB treatment can be improved by adding vitamin D to the pathogenetic treatment, as vitamin D metabolites support the innate immune response to Mycobacterium tuberculosis.&#x0D; OBJECTIVE. To determine the role of vitamin D in the immunopathogenesis of the inflammatory response in pulmonary TB and to assess the prospects of its impact on improving the effectiveness of treatment by analyzing information from available scientific sources on this topic.&#x0D; MATERIALS AND METHODS. The study was performed for the period December 2020 – August 2021. The search was conducted by&#x0D; Keywords:&#x0D; pulmonary tuberculosis, vitamin D, mechanism of action, pathogenesis, treatment. Access to various full-text and abstract databases was used as the main source of research.&#x0D; RESULTS AND DISCUSSION. A large number of studies conducted so far prove the link between vitamin D deficiency and the occurrence of pulmonary TB. Vitamin D receptors have been found to be present on various surfaces of immune cells, including T and B cells, indicating that they need vitamin D to perform cellular functions. Vitamin D has been shown to increase the phagocytic activity of macrophages, and that monocytes incubated with cholecalciferol (vitamin D3) metabolites induce anti-TB activity. A number of studies have shown that vitamin D increases the body’s production of the antimicrobial/antimycobacterial peptide LL-37, a member of the cathelicidin petelide family. Therefore, the narrowly analyzed analysis according to the literature suggests that in the conditions of full vitamin D status of the human body the course of TB will be favorable, and in case of vitamin D deficiency – which is primarily associated with genetic polymorphisms, the course of TB may be unfavorable.&#x0D; CONCLUSIONS. Vitamin D functionates as one of the activators of macrophages and plays a role in the immune defense of the human body against mycobacterial TB. The inclusion of vitamin D in the program of complex treatment of TB infection is promising, as it enhances the production of antimicrobial/antimycobacterial peptide LL-37. It can be used as one of the components of TB prevention in children.

Guttiferone K Exerts the Anti-inflammatory Effect on Mycobacterium Tuberculosis- (H37Ra-) Infected Macrophages by Targeting the TLR/IRAK-1 Mediated Akt and NF-κB Pathway.

Mycobacterium tuberculosis (Mtb) remains a great threat to global health, killing more people than any other single infectious agent and causing uncontrollable inflammation in the host. Poorly controlled inflammatory processes can be deleterious and result in immune exhaustion. The current tuberculosis (TB) control is facing the challenge of drugs deficiency, especially in the context of increasingly multidrug resistant (MDR) TB. Under this circumstance, alternative host-directed therapy (HDT) emerges timely which can be exploited to improve the efficacy of TB treatment and disease prognosis by targeting the host. Here, we established the in vitro infection model of Mtb macrophages with H37Ra strain to seek effective anti-TB active agent. The present study showed that Guttiferone K, isolated from Garcinia yunnanensis, could significantly inhibit Mtb-induced inflammation in RAW264.7 and primary peritoneal macrophages. It was evidenced by the decreased production of inflammatory mediators, including interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2). Further studies with immunoblotting and immunofluorescence revealed that Guttiferone K obviously inhibits the nuclear factor-kappa B (NF-κB) both in RAW264.7 and primary peritoneal macrophages relying on the TLR/IRAK-1 pathway. Guttiferone K could also suppress the NLRP3 inflammasome activity and induce autophagy by inhibiting the protein kinase B (p-Akt) and mammalian target of rapamycin (mTOR) phosphorylation at Ser473 and Ser2448 in both cell lines. Thus, Guttiferone K possesses significant anti-inflammatory effect, alleviating Mtb-induced inflammation with an underlying mechanism that targeting on the TLR/IRAK-1 pathway and inhibiting the downstream NF-κB and Akt/mTOR signaling pathways. Together, Guttiferone K can be an anti-inflammatory agent candidate for the design of new adjunct HDT drugs fighting against tuberculosis.

1886. N1,N12-Diacetylspermine as Potential Urinary Biomarker to Monitor Treatment Response and Bacterial Load in Pulmonary Tuberculosis

BackgroundAdequate control of the global tuberculosis (TB) burden is limited by a lack of accurate measures of treatment efficacy. Current gold standard relies on sputum cultures, which often lag weeks behind time of sample collection. Previous studies have identified urinary N1,N12-diacetylspermine (DiAcSpm) as a potential biomarker of active pulmonary TB (ATB). We aimed to quantify urinary DiAcSpm in participants being treated for ATB and identify its relationship to treatment response.MethodsLongitudinal urine samples were obtained from two separate cohorts of participants treated for ATB. Cohort one consisted of 34 successfully treated ATB cases with urine collected at weeks 0, 2, 4, 8, 17, 26, and 52. Cohort two consisted of 35 ATB cases under two treatment arms: one arm was successfully treated with the standard therapy of rifampin, isoniazid, pyrazinamide, and ethambutol (RIPE), and the other arm received an experimental drug, which was later found to be ineffective against ATB. Urine from cohort two was collected at days 0, 2, 4, and 14. All samples were blinded, randomized, normalized to osmolality and urinary creatinine, and analyzed using high-performance liquid chromatography-mass spectrometry. DiAcSpm concentrations were further tested using ELISA.ResultsUsing linear-mixed modeling that adjusted for age, sex, and participant weight, we found that urinary DiAcSpm significantly decreased by week 2 of successful treatment in both cohorts (P < 0.0001). DiAcSpm levels remained unchanged in the ineffective experimental drug group (cohort two), significantly differentiating treatment success and failure by day 14 (P < 0.0001). Additionally, concentrations of urinary DiAcSpm positively correlate with sputum mycobacterial burden (P = 0.0002).These findings were consistent across both methods of detection.ConclusionOur results suggest that urinary concentrations of DiAcSpm correlate with TB treatment outcome and mycobacterial disease burden, and have the potential to serve as an early biomarker of TB treatment efficacy. Disclosures All Authors: No reported Disclosures.

The Correlation between Body Mass Index and Albumin Level with Sputum Conversion in AFB-Positive Pulmonary TB Patients in Primary Health Center in Medan, Indonesia

Tuberculosis (TB) remains a major public issue in Indonesia including in North Sumatra province. Despite reported good efficacy of TB treatment in the region, the success of treatment depends on many factors including nutritional status. This study aims to determine the relationship between Body Mass Index and Albumin level with sputum conversion in AFB-positive pulmonary TB patients.The study was done in two primary health centres in Medan between October and November 2018. A total of 39 newly diagnosed TB patients with confirmed AFB-positve were included in the study.Participants received TB treatment according to the national guidelines. The proportion of participants with below normal, normal and above normal BMI status were 13 (33.3%), 21 (53.9%) and 5 (12.8%), respectively. Level of albumin was determined as normal in 25 participants (64.1%), and the remaining as low. Normal BMI status was significantly associated with increased albumin level (P&lt;0.05). At 2 months follow-up, the sputum conversion was observed in 24 individuals (61.5%), and the conversion was significantly associated with normal BMI and/or normal albumin level (P&lt;0.05). We concluded that nutritional status is an important factor in the success of TB treatment.

Nutrition Status Related to Clinical Improvement in AFB-Positive Pulmonary Tuberculosis Patients in Primary Health Centres in Medan, Indonesia.

BACKGROUND:Tuberculosis (TB) remains a major public issue in Indonesia, including in North Sumatra province. Despite reported good efficacy of TB treatment in the region, the success of treatment depends on many factors, including nutritional status.AIM:To determine the relationship between Body Mass Index and Albumin level with sputum conversion in AFB-positive pulmonary TB patients.METHODS:The study was done in two primary health centres in Medan between October and November 2018. A total of 39 newly diagnosed TB patients with confirmed AFB-positive were included in the study. Participants received TB treatment according to the national guidelines.RESULTS:The proportion of participants with below normal, normal and above normal BMI status were 13 (33.3%), 21 (53.9%) and 5 (12.8%), respectively. Level of albumin was determined as normal in 25 participants (64.1%), and the remaining as low. Normal BMI status was significantly associated with increased albumin level (p < 0.05). At 2 months follow-up, the sputum conversion was observed in 24 individuals (61.5%), and the conversion was significantly associated with normal BMI and/or normal albumin level (p < 0.05).CONCLUSION:We concluded that nutritional status is an important factor in the success of TB treatment.

ВЛИЯНИЕ ДЕЛЕЦИОННОГО ПОЛИМОРФИЗМА ГЕНОВ GSTM1 И NAT2 НА ПРОГНОЗ ЭФФЕКТИВНОСТИ ЛЕЧЕНИЯ ЧУВСТВИТЕЛЬНОГО ТУБЕРКУЛЕЗА

The aim of the study. To determine the ffect of polymorphism of glutathione-S-transferase class M1 and NAT2 genes on the efficacy of tuberculosis treatment. Material and methods. The genes polymorphism of the xenobiotic detoxification system was determined with polymerase chain reaction. Results. The presence of deletion of the functional zone in GSTM1 gene polymorphic sites increased the risk of low efficacy of tuberculosis treatment and indicated lower chances of resorption of focal-infiltrative changes in the lungs. It was also associated with a low probability of termination of bacterial excretion [OR = 0.07, p = 0.002]. The 0-genotype of the GSTM1 gene was protective against the onset of PDTB. Conclusions. The 0-genotype of the GSTM1 gene is protective against the appearance of tuberculosis. The presence of deletion of the functional zone of the polymorphic site of the GSTM1 gene increases the risk of low efficacy of tuberculosis treatment, is associated with the lowest chances of resorption of focal-infiltrative changes in the lungs, and is accompanied by a low probability of early termination of bacterial excretion.

ЛЕЧЕНИЕ БОЛЬНЫХ ТУБЕРКУЛЕЗОМ И ВИЧ-ИНФЕКЦИЕЙ В СТАЦИОНАРЕ ДНЕВНОГО ПРЕБЫВАНИЯ

Organization of day care centers at TB dispensaries has become a promising trend of directly observed treatment of tuberculosis patients. The objective of the study: to analyze the activities of a day center by TB dispensary for the period of 12 years. Subjects and methods . Two groups of people were compared: patients with TB/HIV co-infection (n = 232) and HIV negative tuberculosis patients (n = 665). Results. HV positive patients had tuberculosis of chest lymph nodes 3.8 times more often. Destructive changes in the lungs were found equally often in both groups in about a quarter of patients. In patients with TB/HIV co-infection, viral hepatitis was diagnosed statistically significantly more often than among HIV negative tuberculosis patients (82.3% versus 11.3%; p &lt; 0.01). Tuberculosis treatment efficacy in HIV positive patients was identical in the day center and in-patient units. Treatment of patients in a dayhospital of TB dispensary has allowed increasing the duration of directly observed treatment of tuberculosis.

Phenotypic Changes on Mycobacterium Tuberculosis-Specific CD4 T Cells as Surrogate Markers for Tuberculosis Treatment Efficacy.

Background: The analysis of phenotypic characteristics on Mycobacterium tuberculosis (MTB)-specific T cells is a promising approach for the diagnosis of active tuberculosis (aTB) and for monitoring treatment success. We therefore studied phenotypic changes on MTB-specific CD4 T cells upon anti-tuberculosis treatment initiation in relation to the treatment response as determined by sputum culture.Methods: Peripheral blood mononuclear cells from subjects with latent MTB infection (n = 16) and aTB (n = 39) at baseline, weeks 9, 12, and 26 (end of treatment) were analyzed after intracellular interferon gamma staining and overnight stimulation with tuberculin. Liquid sputum cultures were performed weekly until week 12 and during 4 visits until week 26.Results: T cell activation marker expression on MTB-specific CD4 T cells differed significantly between subjects with aTB and latent MTB infection with no overlap for the frequencies of CD38pos and Ki67pos cells (both p < 0.0001). At 9 weeks after anti-TB treatment initiation the frequencies of activation marker (CD38, HLA-DR, Ki67) positive MTB-specific, but not total CD4 T cells, were significantly reduced (p < 0.0001). Treatment induced phenotypic changes from baseline until week 9 and until week 12 differed substantially between individual aTB patients and correlated with an individual's time to stable sputum culture conversion for expression of CD38 and HLA-DR (both p < 0.05). In contrast, the frequencies of maturation marker CD27 positive MTB-specific CD4 T cells remained largely unchanged until week 26 and significantly differed between subjects with treated TB disease and latent MTB infection (p = 0.0003).Discussion: Phenotypic changes of MTB-specific T cells are potential surrogate markers for tuberculosis treatment efficacy and can help to discriminate between aTB (profile: CD38pos, CD27low), treated TB (CD38neg, CD27low), and latent MTB infection (CD38neg, CD27high).

First description of agonist and antagonist IP-10 in urine of patients with active TB

ObjectivesBiomarkers for tuberculosis (TB) diagnosis and clinical management are needed to defeat TB. In chronic hepatitis, patients not responding to interferon/ribavirin treatment had high levels of an antagonist form of IP-10. Recently, antagonist IP-10 has been shown to be involved also in TB pathogenesis. Here, we investigated IP-10 agonist/antagonist forms as potential inflammatory biomarkers to support TB diagnosis and monitoring. MethodsTotal IP-10 and its agonist/antagonist forms were measured by SIMOA digital ELISA in urine obtained from patients with active TB at baseline and after treatment. Healthy donors (HD) and patients with pneumonia were enrolled as controls. ResultsPatients with active TB had significantly higher levels of total and agonist IP-10 at baseline compared to HD; conversely, no differences were observed between IP-10 levels in active TB vs pneumonia. Moreover, in active TB a decline of total urine IP-10 was observed at therapy completion; agonist/antagonist forms reflected this decline although their differences were not statistically significant. ConclusionsWe showed for the first time that agonist/antagonist IP-10 forms are measurable in urine. IP-10 levels associate with TB and pneumonia disease, suggesting their association with acute inflammation. Further studies are needed to assess their role to monitor TB treatment efficacy.

Efficacy of T Regulatory Cells, Th17 Cells and the Associated Markers in Monitoring Tuberculosis Treatment Response.

Treatment monitoring is an essential aspect for tuberculosis (TB) disease management. Sputum smear microscopy is the only available tool for monitoring, but it suffers from demerits. Therefore, we sought to evaluate markers and cellular subsets of T regulatory (Treg) cells and T helper (Th) 17 cells in pulmonary TB patients (PTB) for TB treatment monitoring. Peripheral blood mononuclear cells (PBMCs) were stimulated in vitro (with purified protein derivative (PPD)) overnight which was followed by a polychromatic flow cytometry approach to study Treg and Th17 markers and cellular subsets in PTB (n = 12) undergoing antituberculous treatment (ATT). The baseline levels of these markers and cellular subsets were evaluated in normal healthy subjects (NHS). We observed a significant decrease in the expression of CD25 (p<0.01) marker and percentage of T-cell subsets like CD4+CD25+ (p<0.001) and CD4+CD25+CD39+ (p<0.05) at the end of intensive phase (IP) as well as in the continuation phase (CP) of ATT. A decrease in CD25 marker expression and percentage of CD4+CD25+ T cell subset showed a positive correlation to sputum conversion both in high and low sputum positive PTB. In eight PTB with cavitary lesions, only CD4+CD25+FoxP3 Treg subset manifested a significant decrease at the end of CP. Thus, results of this study show that CD25 marker and CD4+CD25+ T cells can serve as better markers for monitoring TB treatment efficacy. The Treg subset CD4+CD25+FoxP3 may be useful for prediction of favorable response in PTB with extensive lung lesions. However, these findings have to be evaluated in a larger patient cohort.

Biodegradable Polymer Nanoparticles of Anti-Tubercular Drug: Formulation and In Vitro Release Studies

Tuberculosis (TB), has been a serious global health problem, resulting in millions of deaths annually. In India itself, 40% of the population are having Latent TB infection and approximately 10% of infected individuals have higher risk of developing active disease, especially those who are immune compromised like HIV infected and patients on immune suppressants.The Latent TB evolves through different stages to become active disease, hence treating latent infection has better scope in controlling TB. Current TB treatment poses several problems and also leads to emergence of drug resistance strains. The biodegradable nanoparticles are promising drug carriers as it has high stability, less toxic and can release the drug in sustained manner. The objective is to provide a new mode of drug delivery through nanoparticles (NP) incorporated with current anti-tubercular drugs that could solve issues with current drug treatment regimen and make available for treatment of latency and active form of TB in long term. In the present study, single emulsion solvent evaporation method was used to formulate and optimize front line anti-tubercular drug with the polymer Poly (lactic co-glycolic acid) PLGA, which is biocompatible and biodegradable. Characterization studies and determination of drug loading efficiency were studied to know the effect of formulation variables. In vitro drug release study was conducted and reverse phase high pressure liquid chromatography method (RP-HPLC) was developed for the analysis of drug. Hence, optimized PLGA, nanoparticles loaded with anti-tuberculosis drug (ATD) have shown good encapsulation efficiency and drug release profile resulted slow and sustained release over a period of several days. In order to target therapeutic molecules, the biodegradable Nano particulate carriers has the potential for improving the efficacy of TB treatment.

Serum levels of soluble urokinase plasminogen activator receptor (suPAR) as a marker of tuberculosis treatment efficacy

BackgroundUpon diagnosis, Pulmonary Tuberculosis patients are treated for TB for a period of 6–9 months. At present, there exists very little indication of the efficacy of the particular treatment. A few previous studies have shown that soluble urokinase plasminogen activator receptor (SuPAR) may be used as treatment efficacy marker. SuPAR is a cellular receptor for serine protease urokinase plasminogen activator (uPA). Bacterial endotoxins and cytokines of the innate immune system stimulate the secretion of uPA in monocytes & neutrophils. Serum SuPAR levels are elevated when TB is active and decrease when the patient responds positively to therapy. ObjectiveTo investigate if SuPAR levels decline upon treatment and whether serum SuPAR levels may be used as a biomarker to monitor Tuberculosis treatment efficacy. DesignThe study was conducted in the department of Biochemistry at VIMS, Ballari, Karnataka. The study subjects were randomly selected from RNTCP centre of VIMS. ControlsTwelve tuberculin skin test positive healthy controls from the community. CasesA total of 60 cases were enrolled for the study and were divided into 3 groups with 20 in each, based on the duration of TB treatment. Group I (n=20): Newly diagnosed pulmonary TB patients before initiation of DOTS. Group II (n=20): TB patients, 2–3 months after initiation of DOTS. Group III (n=20): TB patients who had completed 6 months of DOTS. MethodologyHb%, TC, DC(P)%, DC(L)% & ESR were measured by standard procedures. Serum suPAR was measured by the quantitative sandwich enzyme immunoassay technique using the R & D systems Human uPAR Quantikine ELISA Kit. ResultsThe suPAR levels were elevated before treatment (3.27+2.08ng/ml) and dropped significantly in groups after 2 months of initiation of therapy (2.18+1.17ng/ml) and after completion of 6 months of treatment (1.50+0.93ng/ml). ConclusionThe decrease in suPAR levels in PTB patients with treatment is a manifestation of treatment efficacy. Hence suPAR levels can be used to guide clinical decisions in TB management.

Priority-Setting for Novel Drug Regimens to Treat Tuberculosis: An Epidemiologic Model.

BackgroundNovel drug regimens are needed for tuberculosis (TB) treatment. New regimens aim to improve on characteristics such as duration, efficacy, and safety profile, but no single regimen is likely to be ideal in all respects. By linking these regimen characteristics to a novel regimen’s ability to reduce TB incidence and mortality, we sought to prioritize regimen characteristics from a population-level perspective.Methods and FindingsWe developed a dynamic transmission model of multi-strain TB epidemics in hypothetical populations reflective of the epidemiological situations in India (primary analysis), South Africa, the Philippines, and Brazil. We modeled the introduction of various novel rifampicin-susceptible (RS) or rifampicin-resistant (RR) TB regimens that differed on six characteristics, identified in consultation with a team of global experts: (1) efficacy, (2) duration, (3) ease of adherence, (4) medical contraindications, (5) barrier to resistance, and (6) baseline prevalence of resistance to the novel regimen. We compared scale-up of these regimens to a baseline reflective of continued standard of care.For our primary analysis situated in India, our model generated baseline TB incidence and mortality of 157 (95% uncertainty range [UR]: 113–187) and 16 (95% UR: 9–23) per 100,000 per year at the time of novel regimen introduction and RR TB incidence and mortality of 6 (95% UR: 4–10) and 0.6 (95% UR: 0.3–1.1) per 100,000 per year. An optimal RS TB regimen was projected to reduce 10-y TB incidence and mortality in the India-like scenario by 12% (95% UR: 6%–20%) and 11% (95% UR: 6%–20%), respectively, compared to current-care projections. An optimal RR TB regimen reduced RR TB incidence by an estimated 32% (95% UR: 18%–46%) and RR TB mortality by 30% (95% UR: 18%–44%). Efficacy was the greatest determinant of impact; compared to a novel regimen meeting all minimal targets only, increasing RS TB treatment efficacy from 94% to 99% reduced TB mortality by 6% (95% UR: 1%–13%, half the impact of a fully optimized regimen), and increasing the efficacy against RR TB from 76% to 94% lowered RR TB mortality by 13% (95% UR: 6%–23%). Reducing treatment duration or improving ease of adherence had smaller but still substantial impact: shortening RS TB treatment duration from 6 to 2 mo lowered TB mortality by 3% (95% UR: 1%–6%), and shortening RR TB treatment from 20 to 6 mo reduced RR TB mortality by 8% (95% UR: 4%–13%), while reducing nonadherence to the corresponding regimens by 50% reduced TB and RR TB mortality by 2% (95% UR: 1%–4%) and 6% (95% UR: 3%–10%), respectively. Limitations include sparse data on key model parameters and necessary simplifications to model structure and outcomes.ConclusionsIn designing clinical trials of novel TB regimens, investigators should consider that even small changes in treatment efficacy may have considerable impact on TB-related incidence and mortality. Other regimen improvements may still have important benefits for resource allocation and outcomes such as patient quality of life.