Analytical evaluation of QuantiFERON- Plus and QuantiFERON- Gold In-tube assays in subjects with or without tuberculosis.

The QuantiFERON-TB Gold Plus (QFT-Plus) represents the new QuantiFERON-TB Gold In-tube (QFT-GIT) to identify latent tuberculosis infection (LTBI). The main differences is the addition of a new tube containing shorter peptides stimulating CD8 T-cells. Aim of this study is to evaluate the accuracy of QFT-Plus compared with QFT-GIT in a cross sectional study of individuals with or without tuberculosis (TB). We enrolled 179 participants: 19 healthy donors, 58 LTBI, 33 cured TB and 69 active TB. QFT-Plus and QFT-GIT were performed. The two tests showed a substantial agreement. Moreover we found a similar sensitivity in active TB and same specificity in healthy donors. A higher proportion of the LTBI subjects responded to both TB1 and TB2 compared to those with active TB (97% vs 81%). Moreover, a selective response to TB2 was associated with active TB (9%) and with a severe TB disease, suggesting that TB2 stimulation induces a CD8 T-cell response in absence of a CD4-response. In conclusion, QFT-Plus and QFT-GIT assays showed a substantial agreement and similar accuracy for active TB detection. Interestingly, a higher proportion of the LTBI subjects responded concomitantly to TB1 and TB2 compared to those with active TB, whereas a selective TB2 response associated with active TB.

Lack of Response to HBHA in HIV-Infected Patients with Latent Tuberculosis Infection.

Heparin-binding haemagglutinin (HBHA) has been proposed as an immunological biomarker for discriminating active tuberculosis (TB) from latent TB infection (LTBI) and to identify those at higher risk of progressing to active disease. Few data are available in immune-compromised patients, which are those with increased risk of TB reactivation. The aim of this stusy was to evaluate the immune response to HBHA in HIV-infected subjects with LTBI (HIV-LTBI) or active TB (HIV-TB) in comparison with the immune response to additional Mycobacterium tuberculosis (Mtb) or HIV and CMV antigens. The responses are evaluated in relation to TB status and in the LTBI subjects with the progression to active TB within 2 years. Forty-one HIV-infected antiretroviral-naïve subjects were prospectively enrolled: 18 were HIV-TB and 23 HIV-LTBI. Whole blood was in vitro stimulated overnight with several antigens and mitogen. Interferon-γ response in the harvested plasma was evaluated by ELISA. Despite that CD4 cell count was significantly different between HIV-LTBI and HIV-TB, no differences were observed in response to Mtb- or HIV-specific antigens. Differently, low responses to HBHA were observed in both HIV-LTBI and HIV-TB subjects. Importantly, none of the six HIV-LTBI responding to HBHA developed TB, while two of 17 non-HBHA responders developed active disease. HIV-TB-coinfected subjects, regardless of their TB status, showed low responses to HBHA despite maintaining detectable responses to other antigens; moreover, among the HIV-LTBI, the lack of HBHA responses indicated an increased risk to develop active TB. These results, although preliminary, suggest that a positive response to HBHA in HIV-LTBI correlates with Mtb containment.

Characterization of regulatory T cells identified as CD4(+)CD25(high)CD39(+) in patients with active tuberculosis.

Forkhead box P3 (FoxP3) is a transcription factor whose expression characterizes regulatory T cells (T(reg)), but it is also present on activated T cells, thus hindering correct T(reg) identification. Using classical markers for T(reg) recognition, discordant results were found in terms of T(reg) expansion during active tuberculosis (TB) disease. Recently CD39 has been shown to be an accurate marker for T(reg) detection. The objectives of this study were: (i) to identify T(reg) expressing CD39 in patients with TB and to compare the results with those obtained by the standard phenotypic markers; (ii) to evaluate if T(reg) are expanded in vitro by exogenous interleukin (IL)-2 or by antigen-specific stimulation; and (iii) to characterize T(reg) function on the modulation of antigen-specific responses. We enrolled 13 patients with pulmonary TB and 12 healthy controls. T(reg) were evaluated by flow cytometry ex vivo and after antigen-specific in vitro stimulation using CD25, FoxP3, CD127 and CD39 markers. Results indicate that CD39(+) cells within the CD4(+)CD25(high) cells have T(reg) properties (absence of interferon-gamma production and transforming growth factor-beta1 release upon stimulation). Ex vivo analysis did not show significant differences between TB patients and controls of T(reg) by classical or novel markers. In contrast, a significantly higher percentage of T(reg) was found in TB patients after antigen-specific stimulation both in the presence or absence of IL-2. Depletion of CD39(+) T(reg) increased RD1-specific responses significantly. In conclusion, CD39 is an appropriate marker for T(reg) identification in TB. These results can be useful for future studies to monitor Mycobacterium tuberculosis-specific response during TB.

Mycobacterium tuberculosis DNA detection in soluble fraction of urine from pulmonary tuberculosis patients.

SETTING: A tertiary care and research institution in Italy. BACKGROUND: Small DNA fragments from cells dying throughout the body have been detected in urine (transrenal DNA [Tr-DNA]). OBJECTIVE: To test the hypothesis that Mycobacterium tuberculosis Tr-DNA could be detected in the urine of pulmonary tuberculosis (TB) patients. DESIGN: We studied 43 patients with culture-confirmed pulmonary TB with no evidence of extra-pulmonary involvement, 10 patients with pulmonary diseases other than TB and 13 healthy controls. DNA was extracted from urine and analysed by semi-nested polymerase chain reaction (PCR). RESULTS: M. tuberculosis-specific sequences were found in the urine of 34 of 43 (79%) TB patients studied, whereas all controls were negative. The transrenal nature of M. tuberculosis DNA was demonstrated by two lines of evidence: first, separate analysis of supernatants and sediments from eight of the study patients found seven positive supernatants but only two matched positive sediments. Second, M. tuberculosis-specific sequences were amplified by semi-nested PCR with primers designed for short but not large amplicons. CONCLUSION: Small M. tuberculosis DNA fragments may be detected in the urine of a significant proportion of patients with pulmonary TB. If these observations are confirmed by larger studies, Tr-DNA technology could represent a new approach for detecting pulmonary M. tuberculosis infection.

[Electrocardiographic findings in a school-age population using a computerized system: calculation of standard norms for heart rate and PR, QRS, and QT intervals]. / Rilievi elettrocardiografici in una popolazione in età scolare mediante un sistema computerizzato: calcolo degli standards di normalità per la frequenza cardiaca e gli intervalli P-R, Q-R-S, Q-T.

We evaluated some ECG parameters (HR, P-R, Q-R-S, Q-T) in a healthy school-age population. One-thousand-eight-hundred and ninety children ranging in age from 5 to 12 coming from different zones of the city of Naples were studied. On physical examination all subjects were free from cardiac disease. A computerized ECG (Muse 12SL System Marquette) was performed on every subject. One-hundred and seventy-four of the 1890 children were excluded from statistical analysis because of ectopic rhythm (junctional rhythm or wandering pacemaker) or poor quality of the recording. Of the 1716 children included in the study, 837 were male and 879 were female. The computerized ECG 12SL System Marquette registers an ECG record consisting of all 12 classical ECG leads acquired simultaneously over a 10 second period. Each individual complex can be analyzed in all leads by the computer. An interpretation using this extended record, along with an ECG record of conventional length, is presented to the physician for review. The first step in computerized ECG analysis is Q-R-S identification, then P wave identification, beat classification, rhythm analysis, morphology analysis, complex alignment and computation of median complex. All parameters were divided for sex and age and gathered into tables. The variability of P-R, Q-R-S, Q-T versus HR were also evaluated. The following conclusions were drawn: 1) Sex is a very important variable in the parameters examined. Males have a much slower HR, greater Q-R-S duration, and longer Q-T interval when compared to females. 2) As age increases, HR slowly decreases, while P-R, Q-R-S and Q-T intervals increase.(ABSTRACT TRUNCATED AT 250 WORDS)

Amiodarone treatment in cardiac preexcitation syndrome: use of signal averaged electrocardiogram.

Amiodarone effectiveness to prevent reentrant arrhythmia in Wolff-Parkinson-White (WPW) syndrome is well known. Authors tried to evaluate the results of long-term therapy in a group of 11 patients (mean age 39 +/- years) suffering from WPW syndrome. Before amiodarone treatment, a conventional ECG and a high resolution ECG (a new noninvasive technique) were performed in order to define Hisian activity. After 50 days of therapy (600 mg daily for the first week, 400 mg daily for the second week, 200 mg daily for 5 days in the following period), a second recording revealed the evidence a a lengthening of PR segment (p less than 0.05) and a disappearance of delta wave (1 patient) and arrhythmia. Before treatment, His deflection was defined only in 2 patients. After amiodarone therapy the H-V time was clearly evaluated in 9 patients. Probably the drug has induced a lengthening of AV node refractoriness and primarily an increase of accessory pathway refractoriness.