Flow cytometry-based method using diversity of cytokine production differentiates between Mycobacterium tuberculosis infection and disease.

Authors present a pilot study of the development of innovative flow cytometry-based assay with a potential for use in tuberculosis diagnostics. Currently available tests do not provide robust discrimination between latent tuberculosis infection (TBI) and tuberculosis disease (TB). The desired application is to distinguish between the two conditions by evaluating the production of a combination of three cytokines: IL-2 (interleukin-2), IFNɣ (interferon gamma) and TNFɑ (tumor necrosis factor alpha) in CD4+ and CD8+ T cells. The study was conducted on 68 participants, divided into two arms according to age (paediatric and adults). Each arm was further split into three categories (non-infection (NI), TBI, TB) based on the immune reaction to Mycobacterium tuberculosis (M.tb) after a close contact with pulmonary TB. Each blood sample was stimulated with specific M.tb antigens present in QuantiFERON tubes (TB1 and TB2). We inferred TBI or TB based on the predominant cytokine response of the CD4+ and/or CD8+ T cells. Significant differences were detected between the NI, TBI and the TB groups in TB1 in the CD4+TNFɑ+parameter in children. Along with IL-2, TNFɑ seems to be the most promising diagnostic marker in both CD4+and CD8+ T cells. However, more detailed analyses on larger cohorts are needed to confirm the observed tendencies.

TB index case tracing in the Roma community in the Czech Republic.

Tuberculosis (TB) contact tracing and TB preventive treatment are key tools in preventing the transmission of TB with the aim of eliminating the disease. Our study seeks to demonstrate how the infection spread from an individual patient to the entire community and how proactive contact tracing facilitated prompt diagnosis and treatment. Our work was conducted as a retrospective analysis of the spread of TB infection within the Roma community in the Czech Republic, following the case of an index patient who succumbed to pulmonary TB. Several levels of care and preventive and treatment measures are outlined. Confirming the identity of the Mycobacterium tuberculosis strain was achieved using molecular methods. Among the 39 individuals examined, TB disease was detected in eight patients and TB infection was detected in six patients. The investigation of contacts within this group yielded positive results in 36% of cases, necessitating treatment. The study's findings provide evidence that actively tracing individuals at risk can lead to early detection of cases, prompt treatment, and prevention of further disease transmission. The study also indicates that the highest risk of infection occurs within the sick person's household and that young children under the age of 5 are most susceptible to falling ill.

Manifestations of cutaneous mycobacterial infections in patients with inborn errors of IL-12/IL-23-IFNγ immunity

Background: Inborn errors of IL-12/IL-23-IFNγ immunity underlie Mendelian susceptibility to mycobacterial diseases (MSMD), a group of immunodeficiencies characterized by a highly selective susceptibility to weakly virulent strains of mycobacteria, such as non-tuberculous mycobacteria (NTM) and bacillus Calmette-Guérin (BCG). Cutaneous mycobacterial infections are common in MSMD and may represent a red flag for this immunodeficiency. Objectives: We present a case series of four paediatric patients with MSMD, specifically with IFNγR1 and STAT1 deficiencies, and cutaneous NTM/BCG infections to increase awareness of this immunodeficiency, which may, in some cases, be intercepted by the dermatologist and thus timely referred to the immunologist. Materials & Methods: Clinical, laboratory and genetic investigations of the four paediatric patients with MSMD are presented. Results: All four presented patients experienced early complications after BCG vaccination. Two patients suffered recurrent mycobacteriosis, one patient experienced delayed BCG reactivation, and one patient died of disseminated avian mycobacteriosis. The dermatological manifestation in these patients included destructive nasal ulcerations, scrofuloderma of various sites and lupus vulgaris. All patients had a normal basic immune phenotype. Conclusion: The presented cases demonstrate that NTM/BCG infections in otherwise seemingly immunocompetent patients should raise suspicion of MSMD. This is of utmost importance as specific therapeutic approaches, such as IFNγ treatment or haematopoietic stem cell transplantation, may be employed to improve the disease outcome.

The respiratory consequences of COVID-19 lasted for a median of 4 months in a cohort of children aged 2-18 years of age.

AIM: We focused on the clinical picture, severity and prognosis of children who experienced long-term respiratory issues after COVID-19. METHODS: This was a national Czech multicentre study of paediatric post-COVID syndrome, which used a standard protocol to evaluate structural and functional anomalies and exclude alternative diagnoses. From 6 January to 30 June 2021, 11 paediatric pulmonologists enrolled all paediatric referrals aged 2-18 years with persistent respiratory symptoms more than 12 weeks after COVID-19, namely cough, dyspnoea and chest pain. Medical histories were taken, and physical examinations, lung function testing, chest X-ray and blood tests were performed. RESULTS: The dominant symptoms in the 39 children (56.4% girls) were exertional dyspnoea (76.9%) and a chronic cough (48.7%), while dyspnoea at rest (30.8%) and chest pain (17.9%) were less prevalent. More than half (53.8%) reported more than 1 symptom, and 38.5% had abnormal results for 1 of the following tests: lung function, chest X-ray or D-dimers. The median age of the children was 13.5 years (interquartile range ±4.8 years), and the median recovery time was 4 months (range 1.5-8 months). CONCLUSION: Our initial data suggest that the long-term respiratory impact of COVID-19 was relatively mild in our cohort, with a favourable prognosis.

Trends in incidence of extrapulmonary tuberculosis in children in the Czech Republic in the past 35 years.

OBJECTIVE: Extrapulmonary tuberculosis (EPTB) in children globally represents a proportion of all cases of tuberculosis, reaching 20% according to published reports. Children are at a higher risk for disseminated TB and extrapulmonary forms. The most prevalent clinical presentations of EPTB in children worldwide are peripheral lymphadenitis and osteoarticular TB. Peritoneal, urogenital, or meningeal tuberculosis is less frequent, and their diagnosis is often difficult. The aim of the study was the determination of EPTB incidence in children in a low-prevalence region over 35 years. METHODS: Descriptive retrospective study of the incidence of EPTB in children and adolescents (aged 0-18 years) in the Czech Republic in the years 1987-2021 as reported in the tuberculosis register. RESULTS: Total amount of EPTB cases was 95. The most prevalent form was extrathoracic lymph node TB. The median age was 10 years. Most patients (84%) were Czechs. None of them died. The ratio of pulmonary and extrapulmonary TB was 79.8% to 20.2%. CONCLUSION: The low incidence of EPTB in Czech children is congruent with a low incidence of TB in the Czech Republic. Our study confirms that the most frequent form of EPTB is peripheral lymph node TB. Our study did not prove a significant change in the trend of EPTB after the cessation of the mandatory Bacillus Calmette-Guerin (BCG) vaccination programme.

Ten Years' Experience with the Discontinuation of the Bacillus Calmette-Guérin Vaccination in the Czech Republic.

Background: Bacillus Calmette-Guérin (BCG) vaccination programs were introduced in Czechoslovakia more than 60 years ago under a quite different epidemiological situation than that of today. Compulsory mass BCG vaccination was abolished in November 2010 and changed to a selective vaccination program for infants at high risk of contracting tuberculosis (TB). Methods: This work sets out to ascertain the risk of TB and nontuberculous mycobacterial (NTM) infection in the 10-year period following the change to the vaccination program and to compare this with the same period of time when mass BCG vaccination was compulsory. Descriptive study. Statistical analysis of the incidence of tuberculosis and non-tuberculous mycobacteriosis as reported in the TB register in years 2001-2020. Conclusions: The incidence trend of TB in children in both monitored time periods is identical and statistically significantly decreasing (P < 0.001). In the first monitored period, the incidence trend of NTM cervical lymphadenitis in children is degressive and is not statistically significant (P = 0.561). However, in the second monitored period, the trend increases and is statistically significant (P < 0.001); in every compared 2-consecutive year period, there is an increase of 8%. These findings indicate that the change from mass BCG vaccination to selective vaccination of high-risk newborns and the abolishment of BCG revaccination can be recommended, so long as a keen eye is kept on NTM caused lymphadenitis.

Nontuberculous mycobacterial infections in children in the Czech Republic in the period 2003-2018.

AIM: Comparision of the incidence of cervical lymphadenitis caused by nontuberculous mycobacteria in two equal time periods before and after the ending of widespread calmetization (tuberculosis vaccination). Backgroung. From 2011 to 2018, 89 children were registered in the Tuberculosis Register with cervical lymphadenitis caused by nontuberculous mycobacteria, as confirmed by cultivation. In the majority of cases, the infection was caused by a mycobacterium belonging to the Mycobacterium avium complex. Only 7 cases of cervical lymphadenitis of the same etiology were registered during the same time interval between 2003 and 2010. The authors consider the ending of widespread calmetization (tuberculosis vaccination) in 2010 to be the main cause of the growing incidence. METHOD: A comparison of data for the period 2003-2018 about cases of the illness caused by atypical mycobacteria as reported in the Tuberculosis Register. RESULTS: The average incidence per year in the first interval was 0.04/100 000 children and in the second interval 0.53/100 000 children. During the second time interval, there was an increase from 0.14/100 000 children in 2011 to 1.40/100 000 children in 2018. While the incidence during the first time interval did not show any time trend (P=0.885), the year 2010 marks a significant turning point, with growth during the second interval being highly statistically significant (P<0.001).