Application of IFN-γ/IL-2 FluoroSpot assay for distinguishing active tuberculosis from non-active tuberculosis: A cohort study

Zhang, Lifan; Wan, Shijun; Ye, Susu; Cheng, Xianhao; Zhang, Yueqiu; Shi, Xiaochun; Zhou, Binghua; Sun, Xiaochuan; Liu, Xiaoqing

doi:10.1016/j.cca.2019.08.022

Cited by 4 publications

(7 citation statements)

References 21 publications

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“…The results of the ESAT-6 and CFP-10-Fluorospot revealed that the frequency and proportion of single IFN-γ-secreting T cells in the ATB group were significantly higher than those in the LTBI group ( P = 0.003 and P < 0.001), whereas the frequency and proportion of single IL-2-secreting T cells in the ATB group were significantly lower than those in the LTBI group ( P = 0.004 and P < 0.001), which is consistent with our previous results [15] . The study demonstrated that the combination of the ESAT-6 and CFP-10-FluoroSpot with the Rv1733c SLP resulted in the specificity increased from 66.7% to 83.3%, the positive likelihood ratio increased from 2.47 to 5.05, and the positive predictive value increased from 79.7% to 88.9%.…”

Section: Discussionsupporting

confidence: 92%

“…The cutoff of ESAT-6 and CFP-10-FluoroSpot in the differential diagnosis of ATB and LTBI was defined according to our previous research results. The frequency of single IFN-γ-secreting T cells was 25 (SFCs/2.5 × 10 5 PBMCs), and the proportion of single IFN-γ-secreting T cells was 43.6% [15] . Receiver operating characteristic (ROC) was used to define the best cutoff value of Rv1733c(SLP)-FluoroSpot in the differential diagnosis of ATB and LTBI.…”

Section: Methodsmentioning

confidence: 99%

“…[13] In our previous studies, we confirmed that by using ESAT-6 and CFP-10 peptides as stimulators, the Fluorospot might be used to detect the secretion of IFN-g and interleukin 2 (IL-2), which, in turn, is helpful for the differential diagnosis of ATB and LTBI. [14,15] In this study, we further improved the diagnostic accuracy by combining the MTB latency-associated antigen Rv1733c and Rv1733c SLP based on ESAT-6 and CFP-10-Fluo-roSpot to differentiate ATB from LTBI.…”

Section: Introductionmentioning

confidence: 99%

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Mycobacterium tuberculosis latency-associated antigen Rv1733c SLP improves the accuracy of differential diagnosis of active tuberculosis and latent tuberculosis infection

Zhang

Wan

et al. 2021

Chinese Medical Journal

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Background:Differential diagnosis of active tuberculosis (ATB) and latent tuberculosis infection (LTBI) has been a challenge for clinicians in high TB burden countries. The purpose of this study was to improve the accuracy of differential diagnosis of ATB and LTBI by using fluorescent immunospot (FluoroSpot) assay to detect specific Th1 cell immune responses. The novel mycobacterium tuberculosis (MTB) latency-associated antigens Rv1733c and synthetic long peptides derived from Rv1733c (Rv1733c SLP) were used based on virulence factors early secreting antigen target-6 (ESAT-6) and culture filtrate protein-10 (CFP-10).Methods:Fifty-seven ATB cases, including 20 pathogen-confirmed ATB and 37 clinically diagnosed ATB, and 36 LTBI cases, were enrolled between January and December 2017. FluoroSpot assay was used to detect the interferon γ (IFN-γ) and interleukin 2 (IL-2) secreted by the specific T cells after being stimulated with MTB virulence factors ESAT-6 and CFP-10, MTB latency-associated antigens Rv1733c and Rv1733c SLP. The receiver operating characteristic (ROC) curve was used to define the best cutoff value of latency-associated antigens in the use of differentiating ATB and LTBI. The sensitivity, specificity, predictive value, and likelihood ratio of ESAT-6 and CFP-10-FluoroSpot combined with latency-associated antigen in the differential diagnosis of ATB and LTBI were also calculated.Results:Following the stimulation with Rv1733c and Rv1733c SLP, the frequency of single IL-2-secreting T cells stimulated by Rv1733c SLP had the largest area under the ROC curve, which was 0.766. With a cutoff value of 1 (spot-forming cells [SFCs]/2.5 × 105 peripheral blood mononuclear cells) for frequency, the sensitivity and specificity of distinguishing ATB from LTBI were 72.2% and 73.7%, respectively. ESAT-6 and CFP-10-FluoroSpot detected the frequency and proportion of single IFN-γ-secreting T cells; the sensitivity and specificity of distinguishing ATB from LTBI were 82.5% and 66.7%, respectively. Combined with the frequency of single IL-2-secreting T cells stimulated by Rv1733c SLP on the basis of ESAT-6 and CFP-10-FluoroSpot, the sensitivity and specificity increased to 84.2% and 83.3%, respectively.Conclusion:Rv1733c SLP, combined with ESAT-6 and CFP-10, might be used as a candidate antigen for T cell-based tuberculosis diagnostic tests to differentiate ATB from LTBI.

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Section: Discussionsupporting

confidence: 92%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Mycobacterium tuberculosis latency-associated antigen Rv1733c SLP improves the accuracy of differential diagnosis of active tuberculosis and latent tuberculosis infection

Zhang

Wan

et al. 2021

Chinese Medical Journal

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“…Evidence from previous studies suggests that the IL-2 plays important roles in protective immune responses against MTB infection, and increasing concentrations of IL-2 were observed in the body fluids of active TB individuals compared with control individuals [ 12 ]. However, discordant results concerning the role of IL-2 in predicting the active TB development from LTBI have been reported by various researchers [ 13 , 14 ]. It’s worth noting that the previous conclusions were drawn based on small samples, which could weaken their reliability.…”

Section: Introductionmentioning

confidence: 92%

Combined IFN-γ and IL-2 release assay for detect active pulmonary tuberculosis: a prospective multicentre diagnostic study in China

Tan

Li³

et al. 2021

J Transl Med

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Background We performed a prospective multicentre diagnostic study to evaluate the combined interferon-γ (IFN-γ) and interleukin-2 (IL-2) release assay for detect active pulmonary tuberculosis (TB) in China. Methods Adult patients presenting symptoms suggestive of pulmonary TB were consecutively enrolled in three TB-specialized hospitals. Sputum specimens and blood sample and were collected from each participant at enrolment. The levels of Mycobacterium tuberculosis (MTB)-specific antigen-stimulated IFN-γ and IL-2 were determined using enzyme-linked immunosorbent assay (ELISA). Results Between July 2017 and December 2018, a total of 3245 patients with symptoms suggestive of pulmonary TB were included in final analysis. Of 3245 patients, 2536 were diagnosed as active TB, consisting of 1092 definite TB and 1444 clinically diagnosed TB. The overall sensitivity and specificity of IFN-γ were 83.8% and 81.5%, respectively. In addition, compared with IFN-γ, the specificity of IL-2 increased to 94.3%, while the sensitivity decreased to 72.6%. In addition, the highest sensitivity was achieved with parallel combination of IFN-γ/IL-2, with a sensitivity of 87.9%, and its overall specificity was 79.8%. The sensitivity of series combination test was 68.5%. Notably, the sensitivity of series combination test in definite TB (72.1%) was significantly higher than that in clinically diagnosed TB (65.8%). Conclusion In conclusion, we develop a new immunological method that can differentiate between active TB and other pulmonary diseases. Our data demonstrates that the various IFN-γ/IL-2 combinations provides promising alternatives for diagnosing active TB cases in different settings. Additionally, the diagnostic accuracy of series combination correlates with severity of disease in our cohort.

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“…Results of numerous studies suggest that IL-2 plays important roles in protective immune responses against MTB infection, with detection of increased IL-2 levels in body uids correlating with active TB disease [12]. However, discordant results regarding the utility of IL-2 for predicting active TB development from LTBI have been reported [13,14], although conclusions of numerous studies were drawn based on statistically weak results obtained from studies with small sample sizes. Therefore, additional clinical data is needed to evaluate IL-2 as a marker for differentiating between active TB and LTBI or other respiratory diseases.…”

Section: Introductionmentioning

confidence: 99%

Combined IFN-γ and IL-2 Release Assay for Detect Active Pulmonary Tuberculosis: A Prospective Multicentre Diagnostic Study in China

Yaoju

Tan

et al. 2020

Preprint

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Background: We performed a prospective multicenter diagnostic study to evaluate combined interferon-γ (IFN-γ) and interleukin-2 (IL-2) release assays for detecting active pulmonary tuberculosis (TB) in China. Methods: Adult patients presenting with symptoms suggestive of pulmonary TB were consecutively enrolled in three TB-specialized hospitals. Sputum specimens and blood samples were collected from each participant at enrollment. In vitro measurements of IFN-γ and IL-2 release from Mycobacterium tuberculosis (MTB)-specific antigen-stimulated patient peripheral blood mononuclear cells were conducted using enzyme-linked immunosorbent assays (ELISAs).Results: Between July 2017 and December 2018, a total of 3,245 patients with symptoms suggestive of pulmonary TB were included in the final analysis. Of these patients, 2,536 were diagnosed with active TB, including 1,092 definite and 1,444 clinically diagnosed cases. Overall sensitivity and specificity rates of the IFN-γ release assay were 83.8% and 81.5%, respectively, as compared to IL-2 assay rates yielding greater specificity (94.3%) but lower sensitivity (72.6%). Notably, a parallel-type combination test for IFN-γ/IL-2 provided greatest overall sensitivity (87.9%) but relatively low specificity (79.8%). Meanwhile, a series-type combination test had an overall sensitivity rate of 68.5% that, when stratified by case subgroup, yielded sensitivity rates of 72.1% and 65.8% for definite and clinically diagnosed TB cases, respectively.Conclusion: We developed a new immunological method to differentiate between active TB and other pulmonary diseases. Our data demonstrated that both series-type and parallel-type IFN-γ/IL-2 combination tests may improve diagnosis of active TB cases in different settings. Additionally, the diagnostic accuracy of the series-type combination assay correlated with disease severity in our patient cohort.

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Application of IFN-γ/IL-2 FluoroSpot assay for distinguishing active tuberculosis from non-active tuberculosis: A cohort study

Cited by 4 publications

References 21 publications

Mycobacterium tuberculosis latency-associated antigen Rv1733c SLP improves the accuracy of differential diagnosis of active tuberculosis and latent tuberculosis infection

Mycobacterium tuberculosis latency-associated antigen Rv1733c SLP improves the accuracy of differential diagnosis of active tuberculosis and latent tuberculosis infection

Combined IFN-γ and IL-2 release assay for detect active pulmonary tuberculosis: a prospective multicentre diagnostic study in China

Combined IFN-γ and IL-2 Release Assay for Detect Active Pulmonary Tuberculosis: A Prospective Multicentre Diagnostic Study in China

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