Adult Pulmonary Tuberculosis Research Articles

Delivery of Mycobacterium tuberculosis epitopes by Bordetella pertussis adenylate cyclase toxoid expands HLA-E-restricted cytotoxic CD8+ T cells.

Tuberculosis (TB) remains the first cause of death from infection caused by a bacterial pathogen. Chemotherapy does not eradicate Mycobacterium tuberculosis (Mtb) from human lungs, and the pathogen causes a latent tuberculosis infection that cannot be prevented by the currently available Bacille Calmette Guerin (BCG) vaccine, which is ineffective in the prevention of pulmonary TB in adults. HLA-E-restricted CD8+ T lymphocytes are essential players in protective immune responses against Mtb. Hence, expanding this population in vivo or ex vivo may be crucial for vaccination or immunotherapy against TB. The enzymatically inactive Bordetella pertussis adenylate cyclase (CyaA) toxoid is an effective tool for delivering peptide epitopes into the cytosol of antigen-presenting cells (APC) for presentation and stimulation of specific CD8+ T-cell responses. In this study, we have investigated the capacity of the CyaA toxoid to deliver Mtb epitopes known to bind HLA-E for the expansion of human CD8+ T cells in vitro. Our results show that the CyaA-toxoid containing five HLA-E-restricted Mtb epitopes causes significant expansion of HLA-E-restricted antigen-specific CD8+ T cells, which produce IFN-γ and exert significant cytotoxic activity towards peptide-pulsed macrophages. HLA-E represents a promising platform for the development of new vaccines; our study indicates that the CyaA construct represents a suitable delivery system of the HLA-E-binding Mtb epitopes for ex vivo and in vitro expansion of HLA-E-restricted CD8+ T cells inducing a predominant Tc1 cytokine profile with a significant increase of IFN-γ production, for prophylactic and immunotherapeutic applications against Mtb.

ВОЗРАСТНЫЕ ОСОБЕННОСТИ ФОРМИРОВАНИЯ ВНЕКЛЕТОЧНЫХ ЛОВУШЕК НЕЙТРОФИЛОВ У ЗДОРОВЫХ ЛИЦ И БОЛЬНЫХ ТУБЕРКУЛЁЗОМ

Abstracts. Introduction. Forming a neutrophil extracellular trap (NET) from DNA (NETosis) is considered as a potentially significant marker in the diagnosis of tuberculosis. However, there is practically a lack of data regarding the age-related differences in forming extracellular traps in research publications. Aim. To determine the age-related differences of NETosis in healthy individuals and in tuberculosis (TB) patients. Materials and Methods. The ability of neutrophils to form extracellular traps was investigated in 4 groups: 1) 4-7-year-old children diagnosed with the primary tuberculosis complex, who fell ill within Omsk region; 2) healthy and uninfected children at the same age; 3) adults with tuberculosis, and 4) adults without tuberculosis. For this purpose, isolated fractions of neutrophils from peripheral blood of the groups under research were stimulated with a mixture of bacteria (probiotic) for 30 minutes. In the hanging-drop preparation, we analyzed the proportion of neutrophil extracellular traps and cells in the state of various motility degrees. Results and Discussion. Age-related differences in the ability of neutrophils to form extracellular traps were characterized by a more pronounced response of neutrophils to the NETosis stimulant in children in the control group in comparison with neutrophils isolated from blood in the group of healthy adult volunteers: Neutrophils of children (Me=16.1; Q1=12.8; Q3=18.3) were statistically significantly more intense (p=0.0002) than those of adults (Me=8.6; Q1=7.1; Q3=10.5) and formed filamentous NETs. In the groups of pediatric and adult pulmonary TB patients, the response of neutrophils to the NETosis stimulant was diametrically opposite. Neutrophils of children with primary tuberculosis complex, as of those in the control group, predominantly formed filamentous NETs, but in greater amounts, while neutrophils of adults with infiltrative or focal form and with at most 2 segments involved mainly formed cloudy NETs, whereas filamentous NETs prevailed for the adults in the control group after stimulation. At the same time, proportion of cloudy NETs did not exceed 5% in healthy children and healthy adults. Comparing the findings of our study with literature data, we hypothesized that, most likely, there is a greater increase in PAD4 expression and more pronounced histone H3 citrullination in children’s neutrophils than in those of adults. Conclusions. Our findings indicate more pronounced ex vivo processes induced by NETosis stimulator in both groups of children (healthy individuals and TB patients) as compared to adults in the respective groups, which should be considered by the researchers interpreting the findings of the ability of neutrophils to form extracellular traps in tuberculosis.

Mortality Rates Of Adult Pulmonary Tuberculosis With Malnutrition: A Case Report And Comprehensive Literature Review

Tuberculosis (TB) and malnutrition are still major public health problems in the world. These two clinical conditions can occur simultaneously and create a vicious cycle that worsens TB infection or nutritional status. This case report aims to examine the relationship between low nutritional status (malnutrition) of patients and death due to pulmonary TB. A 31 year old woman came to the pulmonary clinic of a tertiary hospital in Jakarta for her third month of drug-sensitive anti-TB drug (OAT) treatment control. Three months earlier, the patient underwent a rapid diagnostic test (RDT) to determine OAT resistance. The conclusion of her anthropometric examination was body mass index (BMI) of 15.6 kg/m2. Before being diagnosed with pulmonary TB, she always had difficulty gaining weight. During the course of disease, her weight decreased by 5 kg per month. After undergoing treatment for 6 months, the patient was declared complete after 6 months of therapy, without any improvement in nutritional status. At 1 year follow-up, the patient experienced a relapse of pulmonary TB with a BMI of 13.2 kg/m2. She died 2 months after diagnosis of relapse TB. Improvement in clinical complaints and conversion to bacteriological examination are indications of successful therapy, including improvement in nutritional status. To analyze the relationship between malnutrition and pulmonary TB and death, the authors conducted a comprehensive literature search in the journal databases PubMed, Cochrane, and Scopus, followed by analysis according to the PRISMA 2020 flowchart, for a prognostic cohort study of cases of death in pulmonary TB patients with malnutrition. There is a relationship between BMI and the risk of death in TB patients. Anthropometric measurements, especially BMI, need to be carried out during every physical examination of pulmonary TB cases.
 Keywords: Body mass index; Malnutrition; Mortality; Tuberculosis

Intravenous Bacille Calmette–Guérin vaccination protects simian immunodeficiency virus-infected macaques from tuberculosis

Tuberculosis, caused by Mycobacterium tuberculosis (Mtb), is the most common cause of death in people living with human immunodeficiency virus (HIV). Intra-dermal Bacille Calmette–Guérin (BCG) delivery is the only licensed vaccine against tuberculosis; however, it offers little protection from pulmonary tuberculosis in adults and is contraindicated in people living with HIV. Intravenous BCG confers protection against Mtb infection in rhesus macaques; we hypothesized that it might prevent tuberculosis in simian immunodeficiency virus (SIV)-infected macaques, a model for HIV infection. Here intravenous BCG-elicited robust airway T cell influx and elevated plasma and airway antibody titres in both SIV-infected and naive animals. Following Mtb challenge, all 7 vaccinated SIV-naive and 9 out of 12 vaccinated SIV-infected animals were protected, without any culturable bacteria detected from tissues. Peripheral blood mononuclear cell responses post-challenge indicated early clearance of Mtb in vaccinated animals, regardless of SIV infection. These data support that intravenous BCG is immunogenic and efficacious in SIV-infected animals.

Treatment outcome and associated factors among adult patients with pulmonary tuberculosis in selected health centers in Addis Ababa Ethiopia.

The success rate of pulmonary tuberculosis in developing countries is different than expected despite effective treatment. We evaluated treatment outcomes and associated factors of pulmonary tuberculosis patients. A retrospective cross-sectional study was employed among randomly selected health centers in Addis Ababa, Ethiopia. Patient records of adult pulmonary tuberculosis patients treated between January 1st, 2017, and December 31st, 2019 were reviewed. Convenient sampling technique was used to select the study participants. Statistical package for social sciences (SPSS), version 24-computer software was used for analysis. Participants' characteristics were descriptively described, and Bivariate, and multivariate logistic regression analysis were used to determine independent variables related to clinical outcomes. The significance level was determined at p-value < 0.05 and a 95% confidence level. Six hundred thirty-six patient records with a mean age of 37.49± 2.99 were reviewed. The overall treatment success rate was 84.9%. Absence of comorbid illness [AOR = 0.444; 95% CI:0.219-0.900], non-smoking [AOR = 0.35; 95% CI:0.194-0.645], and being HIV negative [AOR = 0.22; 95% CI: 0.106-0.460] were associated with successful treatment outcomes, whereas, not having treatment supporter [AOR = 15.68; 95% CI: 8.11-30.33] was associated with unsuccessful treatment outcome. Treatment success in this study was below the average target set by WHO. HIV positivity, co-morbidities, and smoking increased risk of treatment failure. Patient education about cessation may improve treatment success.

Multi-epitope vaccine against drug-resistant strains of Mycobacterium tuberculosis: a proteome-wide subtraction and immunoinformatics approach.

Mycobacterium tuberculosis (Mtb) is the causative agent of tuberculosis, one of the most deadly infections in humans. The emergence of multidrug-resistant and extensively drug-resistant Mtb strains presents a global challenge. Mtb has shown resistance to many frontline antibiotics, including rifampicin, kanamycin, isoniazid, and capreomycin. The only licensed vaccine, Bacille Calmette-Guerin, does not efficiently protect against adult pulmonary tuberculosis. Therefore, it is urgently necessary to develop new vaccines to prevent infections caused by these strains. We used a subtractive proteomics approach on 23 virulent Mtb strains and identified a conserved membrane protein (MmpL4, NP_214964.1) as both a potential drug target and vaccine candidate. MmpL4 is a non-homologous essential protein in the host and is involved in the pathogen-specific pathway. Furthermore, MmpL4 shows no homology with anti-targets and has limited homology to human gut microflora, potentially reducing the likelihood of adverse effects and cross-reactivity if therapeutics specific to this protein are developed. Subsequently, we constructed a highly soluble, safe, antigenic, and stable multi-subunit vaccine from the MmpL4 protein using immunoinformatics. Molecular dynamics simulations revealed the stability of the vaccine-bound Toll-like receptor-4 complex on a nanosecond scale, and immune simulations indicated strong primary and secondary immune responses in the host. Therefore, our study identifies a new target that could expedite the design of effective therapeutics, and the designed vaccine should be validated. Future directions include an extensive molecular interaction analysis, in silico cloning, wet-lab experiments, and evaluation and comparison of the designed candidate as both a DNA vaccine and protein vaccine.

Factors Influencing Adherence to Pulmonary Tuberculosis Treatment among Adult Patients at St Francis’ Hospital-Katete, Zambia

Background: Pulmonary tuberculosis (PTB) is a contagious bacterial infection of the lungs caused by mycobacterium tuberculosis, also known as tubercle bacilli. The symptoms of PTB can appear gradually and vary in severity. Malnutrition, Human Immunodeficiency Virus (HIV) infection/Acquired Immunodeficiency Syndrome (AIDs), age (young and old), diabetes mellitus, substance abuse, low socioeconomic status, malignancies, and or overcrowding are some of the common risk factors for PTB. According to the Zambia Demographic Health Survey (ZDHS, 2021), Zambia’s prevalence rate of PTB stood at 455 cases per 100,000 population. The World Health Organization (WHO) has recommended Direct Observation of Treatment (DOT) by a trained supervisor (WHO, 2019). However, implementing DOT in Zambia is challenging for both the patient and healthcare provider as it has been observed that a daily visit to a health facility for the first two months by PTB patients was very difficult for a range of reasons which included severe illness at the initiation of treatment, distance too far for walking, and high transportation costs. Objective: The study aimed at determining Factors Influencing Adherence to Pulmonary Tuberculosis Treatment among Adult PTB Patients at St Francis’ Hospital-Katete, Zambia. Methods and materials: Data was collected using a questionnaire and the study sample was one hundred (100) Adult PTB patients, who were selected by simple random sampling method. A quantitative descriptive cross-sectional study design was used. Data was analyzed using Statistical Package for Social Sciences (SPSS) version 21. Chi-square test was used to determine the relationship between independent and dependent variables. Conclusion: The study revealed a significant association between ‘level of information and knowledge, individual motivational support, behavioural skill’ and ‘Adult PTB patients’ adherence to PTB treatment’. Level of Information and knowledge (p-value 0.001), Individual .........

Factors associated with treatment adherence among pulmonary tuberculosis patients in New Delhi

BackgroundTB is treated with a six-month course of four antimicrobial drugs, and nearly all cases of TB can be cured if the medications are given and taken correctly. Due to its prolong treatment plans, there can be reasons associated with non-adherence to treatment by TB patients. Hence, the present study aimed to explore the factors associated with medication adherence among TB patients. MethodA cross-sectional descriptive survey was conducted among adult pulmonary tuberculosis patients enrolled under RNTCP (now NTEP) in New Delhi among 27 functional RNTCP districts. Around 200 TB patients who are enrolled in the Nikshay App and are also on treatment were considered. A structured questionnaire was prepared for the interview guide. Analysis was done using bivariate analysis, chi-square tests, and Fisher's exact tests. ResultsAmong the total participants, 173 (86.5%) were adherent and the remaining 27 (13.5%) participants were non-adherent. The majority of the participants (91%) said they were able to follow the routine to the DOTS center, and 9% said they find it difficult to report to the DOTS center as per their schedule. Only 12.35% of non-adherent participants were seen among those who get regular reminders from their families to take medicines, as compared to 18.42% among those who did not get regular reminders from their families. More than one-fourth of the participants (25.9%) who report not getting necessary motivation from healthcare providers were non-adherent. Motivation by healthcare workers to follow drug schedules was found statistically significant to treatment compliance with a P-value of 0.0422. ConclusionTB is a curable disease; this belief has turned out to be a motivational factor for patients suffering from this disease. Studies have shown that faith in the efficacy of treatment helps adherence to TB treatment while other studies describe how patient adherence was adversely affected by the belief that TB is incurable or the treatment is inefficient or that alternative treatment such as traditional medicine is better.

Identification of Respondent Characteristics and Relationship of Clinical Manifestations with Chest Radiograph Lesion in Pulmonary Tuberculosis Patients at Aloei Saboe Hospital

Introduction: Tuberculosis (TB) is an infectious infectious disease caused by Mycobacterium tuberculosis. This study aimed to determine the relationship between clinical manifestations and the results of chest radiographs in adult pulmonary TB patients at Aloei Saboe Hospital.Method: The research design used in this study was cross-sectional by examining the relationship between clinical manifestations and chest radiography results in adult pulmonary TB patients at Aloei Saboe Hospital from October to November 2022. The population of this study was adult pulmonary TB patients using accidental sampling as a sampling technique. The research instrument used was the medical record which contained the patient's anamnesis status and the results of the chest radiography examination. The relationship between variables was then analyzed using the Kolmogorov-Smirnov test.Results: Of the 44 samples, the most common clinical manifestations were dyspnea and chest pain in 12 patients (26.67%). The most chest radiographic results were fibrotic in 21 patients (47.7%). According to the American Thoracic Association (ATA) classification, extensive lesions were predominantly found in 21 patients (47.7%). There was an association between productive cough and chest radiography results (p=0.012). Meanwhile, chest radiographs were not associated with productive cough, hemoptoe, and dyspnea.Conclusion: There is a relationship between productive cough and chest radiography lesions. Further study may determine samples for research that are minimally biased so that the results obtained are more representative of the population.

Analisis Faktor Risiko TB paru Anak yang Tinggal Serumah dengan Penderita TB paru Dewasa

Pulmonary tuberculosis (TB) is still a major public health problem in Indonesia. Based on the Household Health Survey (SKRT) results tuberculosis is the number one respiratory disease and infectious disease. Tuberculosis in children causes growth disorders and even death. If pulmonary tuberculosis in a child is not treated immediately, it can quickly develop into parenchymal or tubercular pneumonia, milliary tuberculosis, bone tuberculosis (scrofuloderma), joint tuberculosis, abdominal tuberculosis, and even meningitis. This study used a case-control design with analytical observation. Children who were exposed to and lived in the same household with adult pulmonary tuberculosis patients as many as 90 adults participated in the study. The case sample consisted of children aged 0-15 years who were diagnosed with Pulmonary TB by doctors at the Wania Health Center, Mimika Regency. The case sample size was 30 respondents and the control sample was 30 respondents. The free variables in this study were parental knowledge, BCG vaccination history, child nutritional status, occupancy density, the behavior of TB sufferers at home, and length of contact. Data were analyzed using univariate analysis, bivariate analysis, and multivariate analysis (multiple logistic regression). The results of the study analysis showed that the factors that influence the incidence of tuberculosis in children are parental knowledge (p = 0.001), BCG vaccination history (p = 0.001), and the behavior of adult tuberculosis sufferers in the household (p = 0.000) The conclusion of the study was that the behavior of adult pulmonary tuberculosis patients who lived in the same house with children was most influential in the transmission of pulmonary tuberculosis in children who lived in the same household as adult pulmonary tuberculosis patients at the Health Centre of Wania, Mimika District.

Xpert MTB/RIF Ultra Assay Using Stool: an Effective Solution for Bacilli Identification from Adult Pulmonary Tuberculosis Suspects without Expectorated Sputum.

This study evaluated the diagnostic performance of stool-based Xpert MTB/RIF Ultra assay (Xpert-Ultra, Cepheid, USA) against other tests using respiratory tract specimens (RTS) and stool for diagnosing adult pulmonary tuberculosis. A prospective study on patients with presumptive pulmonary tuberculosis was conducted in Beijing Chest Hospital from June to November 2021. The smear test, MGIT960 liquid culture, and Xpert MTB/RIF (Xpert, Cepheid, USA) were performed simultaneously on RTS, and smear, culture Xpert, and Xpert-Ultra were performed simultaneously using stool. Patients were grouped based on the outcomes of RTS examination and other tests. In total, 130 eligible patients were enrolled that included 96 pulmonary tuberculosis and 34 non-TB patients. The sensitivity of smear, culture, Xpert, and Xpert-Ultra using stool was 10.96%, 23.28%, 60.27%, and 79.45%, respectively. The specificities of Xpert and Xpert-Ultra using RTS and stool were all 100% (34/34). Notably, all five confirmed cases detected by bronchoalveolar lavage fluid (BALF) examination yielded Xpert-Ultra positive outcomes with the stool specimens. Xpert-Ultra assay on stool sample harbors comparable sensitivity with Xpert on RTS. Thus, the Xpert-Ultra testing on stool specimens could be a very promising and practical strategy to improve pulmonary tuberculosis (PTB) diagnosis, especially among patients who could not expectorate sputum. IMPORTANCE This study is aimed at assessing the value of Xpert MTB/RIF Ultra (Xpert-Ultra) in PTB on stool in adult in low HIV settings and Xpert-Ultra assay on stool sample harboring comparable sensitivity with Xpert MTB/RIF on respiratory tract specimens. Although the yield in stool samples by Xpert-Ultra is lower than RTS, it may be useful in detecting disease in presumptive TB patients who cannot expectorate sputum and are not open to BALF collection. In addition, Xpert-Ultra with a "trace call" on stool in adult was highly supportive of PTB.

Long-Term Intake of Linezolid Elevates Drug Exposure and Reduces Drug Clearance and Elimination in Adults With Drug-Resistant Pulmonary Tuberculosis.

Pharmacokinetic (PK) studies are critical for dose optimization, and there is a paucity of linezolid (LZD) PK data for prolonged use in drug-resistant tuberculosis (DR-TB). Therefore, the authors evaluated the pharmacokinetics of LZD at two-time intervals in DR-TB during long-term use. PK evaluation of LZD was performed at the end of the 8th and 16th weeks of treatment in a randomly selected subset of adult pre-extensively drug-resistant pulmonary tuberculosis patients (n = 18) from a multicentric interventional study (Building Evidence to Advance Treatment of TB/BEAT study; CTRI/2019/01/017310), wherein a daily dose of 600 mg LZD was used for 24 weeks. Plasma LZD levels were measured using a validated high-pressure liquid chromatography (HPLC) method. The LZD median plasma C max was comparable between the 8th and 16th weeks [18.3 mg/L, interquartile range (IQR: 15.5-20.8 and 18.8 mg/L, IQR: 16.0-22.7, respectively)]. However, the trough concentration increased significantly in the 16th week (3.16 mg/L, IQR: 2.30-4.76), compared with the 8th week (1.98 mg/L, IQR: 0.93-2.75). Furthermore, compared with the 8th week, in the 16th week, there was a significant increase in drug exposure (AUC 0-24 = 184.2 mg*h/L, IQR: 156.4-215.8 versus 233.2 mg*h/L, IQR: 187.9-277.2), which corroborated with a longer elimination half-life (6.94 hours, IQR: 5.55-7.99 versus 8.47 hours, IQR:7.36-11.35) and decreased clearance (2.91 L/h, IQR: 2.45-3.33 versus 2.19 L/h, IQR: 1.49-2.78). Long-term daily intake of 600 mg LZD resulted in a significant elevation in trough concentration (>2.0 mg/L) in 83% of the study participants. Furthermore, increased LZD drug exposure may be partly because of decreased clearance and elimination. Overall, the PK data underscore the need for dose adjustment when LZDs are intended for long-term treatment.

Burden of Pulmonary Rifampicin-Resistant Tuberculosis in Kajiado, Kenya: An Observational Study.

Rifampicin resistance (RR) is a major challenge in the clinical management of tuberculosis (TB), but data on its prevalence are still sparse in many countries. Our study aimed at estimating the prevalence of RR-TB in Kajiado County, Kenya. Secondary objectives were to estimate the incidence of pulmonary TB in adults and the rate of HIV-TB coinfection. We conducted an observational study in the context of the ATI-TB Project, carried out in Kajiado. The project was based on an active-case-finding campaign implemented with the aid of village chiefs, traditional healers and community health volunteers. Diagnosis relied on Xpert MTB/RIF, including a mobile machine that could be used to cover areas where testing would otherwise be difficult. In sum, 3840 adults were screened for active TB during the campaign. RR cases among all TB diagnoses were 4.6%. The annual incidence of pulmonary TB among adults was 521 cases per 100,000 population. The rate of HIV coinfection was 22.2% among pulmonary TB diagnoses. The prevalence of RR-TB was four times that what could be inferred from official notifications in Kajiado, and higher than overall prevalence in Kenya. In addition, our estimate of incidence of pulmonary TB in adults in Kajiado significantly differed from cases notified in the same area. In contrast, the rate of HIV coinfection was in line with national and regional data. TB diagnostic capability must be strengthened in Kajiado to improve patients' management and public health interventions.

Comparative serum lipid and immunohematological values among adult pulmonary tuberculosis and tuberculosis lymphadenitis cases and their association with sputum bacilli load and time to culture positivity in Northwestern Ethiopia

BackgroundThe serum lipid and immunohematological values of tuberculosis lymphadenitis (TBLN) patients is poorly documented relative to pulmonary tuberculosis (PTB) cases. Therefore, the aim of this study was to investigate the serum lipid and immunohematological values of patients with TBLN in comparison with PTB (PTB) patients.MethodsAn institution-based comparative cross-sectional study was conducted in Northwest Ethiopia from March to December 2021. The study participants were bacteriologically confirmed PTB (n = 82) and TBLN (n = 94) cases with no known comorbidity and whose ages was greater than 18 years and with no current pregnancy. Independent sample t-test, one-way ANOVA, box plot, and correlation matrix were used to analyze the data.ResultsThe body mass index (BMI), CD4 + T cell count, and high-density lipoprotein-Cholesterol (HDL-C) values were significantly higher among TBLN cases compared with PTB cases. Additionally, the total white blood cell (WBC) count, hemoglobin (Hb), total Cholesterol (CHO) and creatinine (Cr) values were relatively higher among TBLN than PTB (P > 0.05). On the reverse, the platelet count and triacylglycerol (TAG) values were relatively higher among PTB than in TBLN cases. While the mean days of culture positivity were 11.6 days for TBLN, the mean days of culture positivity were 14.0 days for PTB. Anemia and serum lipid values showed no correlation with sputum bacilli load and time to culture positivity.ConclusionTuberculous lymphadenitis patients were well-endowed with serum lipid, immunological and nutritional status compared with PTB cases. Hence, the high incidence rate of TBLN in Ethiopia could not be explained by low peripheral immunohematological values, malnutrition, Anemia, and dyslipidemia. Further study for identifying the predictors for TBLN in Ethiopia is highly desirable.

Synthesis of plant-based, self-adjuvanted, dual antigen specific to Mycobacterium tuberculosis as a novel tuberculosis subunit vaccine that elicits immunogenicity in rabbit.

The only approved vaccine, Bacillus Calmette Guérin (BCG) used in global tuberculosis (TB) immunization programmes has been very effective in childhood TB but not in adult pulmonary and latent TB. Moreover, the emergence of multi-drug resistance-TB cases demands either to increase efficiency of BCG or replace it with the one with improved efficacy. A novel combination of two most effective secreted protein antigens specific for Mycobacterium tuberculosis (Mtb), ESAT-6 and MPT-64 (but not present in BCG strains) fused with a cholera toxin B subunit (CTB) and tagged with 6xHis was expressed for the first time in Escherichia coli as well as in transgenic cucumber plants developed using Agrobacterium tumefaciens-mediated transformation. The recombinant fusion protein (His6x.CTB-ESAT6-MPT64) expressed in E. coli was purified by a single-step affinity chromatography and used to produce polyclonal antibodies in rabbit. The transgenic cucumber lines were confirmed by polymerase chain reaction (PCR), Southern blot hybridization, reverse transcriptase PCR (RT-PCR), real-time PCR (qRT-PCR) and expression of recombinant fusion protein by western blot analysis and its quantification by enzyme-linked immunosorbent assay (ELISA). A maximum value of the fusion protein, 478ng.g-1 (0.030% of the total soluble protein) was obtained in a transgenic cucumber line. Rabbit immunized orally showed a significant increase in serum IgG levels against the fusion protein as compared to the non-immunized rabbit. Stable expression of Mtb antigens with CTB in edible cucumber plants (whose fruits are eaten raw) in sufficient amount possibly would facilitate development of a safe, affordable and orally delivered self-adjuvanted, novel dual antigen based subunit vaccine against TB.

Diabetes mellitus is associated with an increased risk of unsuccessful treatment outcomes among drug-susceptible tuberculosis patients in Ethiopia: A prospective health facility-based study

BackgroundThe impact of diabetes mellitus on tuberculosis (TB) treatment outcomes has not been well investigated in most sub-Saharan countries including Ethiopia. The current study aimed to determine the association between diabetes mellitus and unsuccessful TB treatment outcomes among drug-susceptible TB patients treated at selected health facilities in Addis Ababa, Ethiopia. MethodsThis health facility-based prospective cohort study was conducted at six randomly selected public health centers in Addis Ababa, from August 2020 until November 2021. Clinically diagnosed adult pulmonary and extra pulmonary TB patients were recruited at the time of treatment initiation. A multivariable logistic regression analysis was used to estimate the association between diabetes and unsuccessful TB treatment outcomes. ResultsAmong the total 267 enrolled participants, 9.7% of patients with TB were identified to have diabetes comorbidity. Of patients with diabetes and TB, 9 (34.6%) were newly diagnosed based on glucose test results. Despite an overall high TB treatment success rate (94.0%), more than one-fourth (26.9%) of patients with diabetes had a poor TB treatment outcome (26.9%), which was remarkably higher compared to patients without diabetes (3.7%). In multivariable regression, the adjusted odds of poor TB treatment outcome among those with diabetes was 14.8 (95% CI 3.5 – 62.7) times the odds of poor outcome patients without diabetes. ConclusionDiabetes was significantly associated with increased odds of poor TB treatment outcomes among patients in Addis Ababa, Ethiopia.

SIRT2 inhibition by AGK2 enhances mycobacteria-specific stem cell memory responses by modulating beta-catenin and glycolysis

Bacille Calmette-Guerin (BCG) generates limited long-lasting adaptive memory responses leading to short-lived protection against adult pulmonary tuberculosis (TB). Here, we show that host sirtuin 2 (SIRT2) inhibition by AGK2 significantly enhances the BCG vaccine efficacy during primary infection and TB recurrence through enhanced stem cell memory (TSCM) responses. SIRT2 inhibition modulated the proteome landscape of CD4+ Tcells affecting pathways involved in cellular metabolism and T-cell differentiation. Precisely, AGK2 treatment enriched the IFNγ-producing TSCM cells by activating β-catenin and glycolysis. Furthermore, SIRT2 specifically targeted histone H3 and NF-κB p65 to induce proinflammatory responses. Finally, inhibition of the Wnt/β-catenin pathway abolished the protective effects of AGK2 treatment during BCG vaccination. Taken together, this study provides a direct link between BCG vaccination, epigenetics, and memory immune responses. We identify SIRT2 as a key regulator of memory Tcells during BCG vaccination and project SIRT2 inhibitors as potential immunoprophylaxis against TB.

The radiologic findings, positivity rate of culture method examinations, correlation with the type of lower respiratory secretion of adult pulmonary Tuberculosis (TB) patients

Background: There are active and latent tuberculosis types, a global public health concern. Test confirmation is required to distinguish between active tuberculosis and nontuberculous mycobacterial infection. This study aims to evaluate the relationship between the type of lower airway secretion specimen, Mycobacterium tuberculosis positivity rate of MGIT-LJ, as well as the relationship between the Mycobacterium tuberculosis positivity rate in MGIT-LJ and radiological findings. Methods: This study is cross-sectional with a prospective design. The sample is the result of the examination of the MGIT-LJ method and medical record data of patients suspected of Pulmonary TB at Dr. Soetomo Academic Hospital Surabaya Indonesia that meets the inclusion criteria, analysis of the culture results of MGIT-LJ method and radiological findings were carried out to assess the relationship between the character of sputum, the positivity rate of Mycobacterium tuberculosis and radiological findings. Result: A total of 137 from June 2022 till November 2022 found 88 (64.2%) with spontaneous sputum, 6 (4,4%) aspirate ETT, and 43 (31.4%) BAL. The positivity rate of the MGIT-LJ culture method of the spontaneous sputum specimen to detect Mycobacterium tuberculosis was 20.5% higher than ETT or BAL specimens because the implementation of clinical indication to request MGIT-LJ culture method of ETT and BAL specimens is still confusing. The specificity of the chest radiography diagnostic test from all specimens type of 61.86%, with a weak accuracy rate. Conclusion: In this study, the positivity rate of the MGIT-LJ culture method of the spontaneous sputum specimen to detect Mycobacterium tuberculosis was 20.5% higher than ETT or BAL specimens. Chest radiography has a weak accuracy level and must be complemented test with the culture method.

Stool specimen for diagnosis of pulmonary tuberculosis in adults: a systematic review

ObjectiveTo assess the diagnostic accuracy of stool specimens to diagnose pulmonary tuberculosis (PTB) in adults.DesignSystematic review.Data sourcesMEDLINE (Ovid), Embase (Ovid), Web of Science and the Cochrane database were searched from...

The natural history of untreated pulmonary tuberculosis in adults: a systematic review and meta-analysis.

Stages of tuberculosis disease can be delineated by radiology, microbiology, and symptoms, but transitions between these stages remain unclear. In a systematic review and meta-analysis of studies of individuals with untreated tuberculosis who underwent follow-up (34 cohorts from 24 studies, with a combined sample of 139 063), we aimed to quantify progression and regression across the tuberculosis disease spectrum by extracting summary estimates to align with disease transitions in a conceptual framework of the natural history of tuberculosis. Progression from microbiologically negative to positive disease (based on smear or culture tests) in participants with baseline radiographic evidence of tuberculosis occurred at an annualised rate of 10% (95% CI 6·2-13·3) in those with chest x-rays suggestive of active tuberculosis, and at a rate of 1% (0·3-1·8) in those with chest x-ray changes suggestive of inactive tuberculosis. Reversion from microbiologically positive to undetectable disease in prospective cohorts occurred at an annualised rate of 12% (6·8-18·0). A better understanding of the natural history of pulmonary tuberculosis, including the risk of progression in relation to radiological findings, could improve estimates of the global disease burden and inform the development of clinical guidelines and policies for treatment and prevention.