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

Abstract Background The differential diagnosis of active tuberculosis(ATB) and latent tuberculosis infection(LTBI) is still challenging. The objective of the study was to evaluate the accuracy of the novel M. tuberculosis latency-associated antigens Rv1733c and Rv1733c SLP for differentiating ATB from LTBI. Methods A case-control study was designed to enroll pathogen-confirmed ATB cases admitted to the Peking Union Medical College Hospital and Beijing Chest Hospital, whereas those with LTBI were denoted as the control group. The Fluorescence-Immunospot (FluoroSpot) assay was used to detect the frequencies of IL-2-, IFN-γ-secreting T cells stimulated by the M. tuberculosis latency-associated antigens Rv1733c and Rv1733c SLP. The combination of the ESAT-6/CFP-10-Fluorospot test was evaluated with regard to the sensitivity, specificity, predictive value and likelihood ratio for the differential diagnosis of ATB and LTBI. Results A total of 20 pathogens-confirmed TB and 28 LTBI cases were included. The sensitivity and specificity of ESAT-6/CFP-10-FluoroSpot for the differential diagnosis of ATB and LTBI were 95% (95% CI, 75.13–99.87%) and 82.14% (95% CI, 63.11–93.94%), respectively. Following stimulation with Rv1733c and Rv1733c SLP, the maximum AUROC was 0.711 (95% CI, 0.566–0.856) as determined by the ROC curve, which was used to assess the frequency of single IL-2-secreting T cells stimulated by Rv1733c SLP. The cutoff value of 0 SFCs/2.5 × 105 PBMCs was used for the analysis. The frequency, sensitivity and specificity of Rv1733c SLP for differentiating ATB and LTBI were 75% (95% CI, 50.90–91.34%) and 60.71% (95% CI, 40.58–78.50%), respectively. The ESAT-6/cfp-10-fluorospot was combined with the frequency of single IL-2-secreting T cells, which were stimulated by Rv1733c SLP for the differential diagnosis of ATB and LTBI. This resulted in an increased sensitivity and specificity to 100% (95% CI, 83.16–100.00%), as determined by the parallel test and to 92.86% (95% CI, 71.77–97.73%) as determined by the serial test, respectively. Conclusions Rv1733c SLP has the potential to be used as a candidate antigen for T cell-based tuberculosis diagnostic tests, in combination with ESAT-6 and CFP-10, to differentiate between ATB and LTBI diagnosis.

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Mycobacterium tuberculosis Secreted Proteins As Potential Biomarkers for the Diagnosis of Active Tuberculosis and Latent Tuberculosis Infection

Journal of Clinical Laboratory Analysis ◽

10.1002/jcla.21782 ◽

2014 ◽

Vol 29 (5) ◽

pp. 375-382 ◽

Cited By ~ 25

Author(s):

Caiqin Zhang ◽

Xiaoqin Song ◽

Yong Zhao ◽

Hai Zhang ◽

Shanmin Zhao ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Latent Tuberculosis Infection ◽

Latent Tuberculosis ◽

Active Tuberculosis ◽

Tuberculosis Infection ◽

Secreted Proteins ◽

Potential Biomarkers

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Activation Phenotype of Mycobacterium tuberculosis-Specific CD4+ T Cells Promoting the Discrimination Between Active Tuberculosis and Latent Tuberculosis Infection

Frontiers in Immunology ◽

10.3389/fimmu.2021.721013 ◽

2021 ◽

Vol 12 ◽

Author(s):

Ying Luo ◽

Ying Xue ◽

Liyan Mao ◽

Qun Lin ◽

Guoxing Tang ◽

...

Keyword(s):

T Cells ◽

Mycobacterium Tuberculosis ◽

Latent Tuberculosis Infection ◽

Latent Tuberculosis ◽

Active Tuberculosis ◽

Tuberculosis Infection ◽

Antigen Stimulation ◽

Hla Dr ◽

Tnf Α ◽

Ifn Γ

BackgroundRapid and effective discrimination between active tuberculosis (ATB) and latent tuberculosis infection (LTBI) remains a challenge. There is an urgent need for developing practical and affordable approaches targeting this issue.MethodsParticipants with ATB and LTBI were recruited at Tongji Hospital (Qiaokou cohort) and Sino-French New City Hospital (Caidian cohort) based on positive T-SPOT results from June 2020 to January 2021. The expression of activation markers including HLA-DR, CD38, CD69, and CD25 was examined on Mycobacterium tuberculosis (MTB)-specific CD4+ T cells defined by IFN-γ, TNF-α, and IL-2 expression upon MTB antigen stimulation.ResultsA total of 90 (40 ATB and 50 LTBI) and another 64 (29 ATB and 35 LTBI) subjects were recruited from the Qiaokou cohort and Caidian cohort, respectively. The expression patterns of Th1 cytokines including IFN-γ, TNF-α, and IL-2 upon MTB antigen stimulation could not differentiate ATB patients from LTBI individuals well. However, both HLA-DR and CD38 on MTB-specific cells showed discriminatory value in distinguishing between ATB patients and LTBI individuals. As for developing a single candidate biomarker, HLA-DR had the advantage over CD38. Moreover, HLA-DR on TNF-α+ or IL-2+ cells had superiority over that on IFN-γ+ cells in differentiating ATB patients from LTBI individuals. Besides, HLA-DR on MTB-specific cells defined by multiple cytokine co-expression had a higher ability to discriminate patients with ATB from LTBI individuals than that of MTB-specific cells defined by one kind of cytokine expression. Specially, HLA-DR on TNF-α+IL-2+ cells produced an AUC of 0.901 (95% CI, 0.833–0.969), with a sensitivity of 93.75% (95% CI, 79.85–98.27%) and specificity of 72.97% (95% CI, 57.02–84.60%) as a threshold of 44% was used. Furthermore, the performance of HLA-DR on TNF-α+IL-2+ cells for differential diagnosis was obtained with validation cohort data: 90.91% (95% CI, 72.19–97.47%) sensitivity and 68.97% (95% CI, 50.77–82.73%) specificity.ConclusionsWe demonstrated that HLA-DR on MTB-specific cells was a potentially useful biomarker for accurate discrimination between ATB and LTBI.

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Diagnostic Potential of IgG and IgA Responses to Mycobacterium tuberculosis Antigens for Discrimination among Active Tuberculosis, Latent Tuberculosis Infection, and Non-Infected Individuals

Microorganisms ◽

10.3390/microorganisms8070979 ◽

2020 ◽

Vol 8 (7) ◽

pp. 979

Author(s):

Ji Yeon Lee ◽

Byoung-Jun Kim ◽

Hyeon-Kyoung Koo ◽

Junghyun Kim ◽

Jee-min Kim ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Latent Tuberculosis Infection ◽

Latent Tuberculosis ◽

Public Health Problem ◽

Active Tuberculosis ◽

Antibody Responses ◽

Chorismate Mutase ◽

Tuberculosis Infection ◽

Major Public Health Problem ◽

Diagnostic Potential

Tuberculosis remains a major public health problem. Conventional tests are inadequate to distinguish between active tuberculosis (ATB) and latent tuberculosis infection (LTBI). We measured antibody responses to Mycobacterium tuberculosis antigens (Mycobacterium tuberculosis chorismate mutase (TBCM), antigen 85B (Ag85B), early secreted antigen-6 (ESAT-6), and culture filtrate protein-10 (CFP-10) in ATB, LTBI, and non-infected (NI) individuals. Serum immunoglobulin G (IgG) and immunoglobulin A (IgA) levels were measured and the QuantiFERON-TB Gold In-Tube assay was used to diagnose LTBI. IgG levels against TBCM were significantly higher in LTBI than NI subjects. IgG and IgA levels against Ag85B and IgG levels against CFP-10 were significantly higher in ATB, followed by LTBI, and then NI. When the ATB group was subdivided, IgG levels against Ag85B and CFP-10 were significantly higher in each subgroup compared with those in LTBI and NI groups. Positive correlation trends between interferon-gamma and IgG levels against Ag85B, TBCM, and CFP-10 and IgA levels against Ag85B in LTBI and NI subjects were observed. Age- and sex-adjusted models showed that IgG against TBCM and CFP-10 was independently related to LTBI diagnosis, and IgG against Ag85B was independently related to the diagnosis of ATB and could distinguish between LTBI and ATB. Overall, IgG antibody responses to TBCM, Ag85B, and CFP-10 can discriminate among ATB, LTBI, and NI groups.

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1405. The Accuracy of Mycobacterium tuberculosis Specific IFN-γ/IL-2/TNF-α- FluoroSpot in Differential Diagnosis of Active Tuberculosis and Latent Tuberculosis Infection: A Case-Control Study

Open Forum Infectious Diseases ◽

10.1093/ofid/ofab466.1597 ◽

2021 ◽

Vol 8 (Supplement_1) ◽

pp. S786-S787

Author(s):

Lifan Zhang ◽

Huimin Ma ◽

Qiping Ge ◽

Yueqiu Zhang ◽

Xiaochun Shi ◽

...

Keyword(s):

T Cells ◽

Differential Diagnosis ◽

Mycobacterium Tuberculosis ◽

Latent Tuberculosis Infection ◽

Specific Antigen ◽

Latent Tuberculosis ◽

Active Tuberculosis ◽

Tnf Α ◽

Ifn Γ ◽

Stimulation Of

Abstract Background To establish the Mycobacterium tuberculosis (MTB) specific IFN-γ/IL-2/TNF-α-FluoroSpot assay, and preliminarily evaluate its accuracy of differential diagnosis of active tuberculosis (ATB) and latent tuberculosis infection (LTBI). Methods Patients with pathologically confirmed and clinically diagnosed ATB in Peking Union Medical College Hospital and Beijing Chest Hospital from April 2020 to May 2021 were enrolled as case group, while patients with LTBI in the same period were enrolled as control group. The FluoroSpot assay was used to simultaneously detect the secretion of IFN-γ, IL-2 and TNF-α in T cells stimulated by the MTB specific antigens ESAT-6 and CFP-10 at the single-cell level. A binary logistic regression model was used to fit the combined diagnostic parameters, and the sensitivity, specificity, predictive value and likelihood ratio of the differential diagnosis of ATB and LTBI were calculated. Figure 1. Schematic diagram of FluoroSpot (IFN-γ/IL-2/TNF-α) detecting cytokine-secreting specific T cells after stimulation with MTB specific antigen. A. The green spots are the total IFN-γ-secreting T cells; B. The red spots are the total IL-2-secreting T cells; C. The blue spots are the total TNF-α-secreting T cells; D. The green spots are the single IFN-γ-secreting T cells; the red spots are the single IL-2-secreting T cells; the blue spots are the single TNF-α-secreting T cells; the yellow spots are the dual IFN-γ/IL-2-secreting T cells; the cyan spots are the dual IFN-γ/TNF-α-secreting T cells; the purple spots are the dual IL-2/TNF-α-secreting T cells; the white spots are the triple IFN-γ/IL-2/TNF-α-secreting T cells. Results 62 patients with ATB (37 pathogen-confirmed ATB, 25 clinical diagnosed ATB), 87 patients with LTBI were included. There was significant correlation of the frequencies of total IFN-γ-secreting T cells detected by IFN-γ/IL-2/TNF-α-FluoroSpot assay compared with T-SPOT.TB after stimulation of MTB-specific antigen (r=0.829 for ESAT-6, P< 0.001, r=0.804 for CFP-10, P< 0.001). ROC curve was drawn for both T-SPOT.TB and Fluorospot. For T-SPOT.TB, the AUROC was 0.669 (95%CI 0.574-0.765), the sensitivity and specificity of differentiating ATB from LTBI were 70.97% (95%CI 58.05%-81.80%) and 56.32% (95CI 45.26%-66.94%) respectively. While for Fluorospot, the AUROC was 0.906 (95 CI 0.856-0.957), the sensitivity and specificity of differentiating ATB from LTBI were 80.65% (95%CI 68.63% - 89.58%) and 88.51% (95%CI 79.88% - 94.35%) respectively. Figure 2. Correlation between the frequencies of total IFN-γ-secreting T cells detected by FluoroSpot assay and those of T-SPOT.TB. (A) Stimulated by EAST-6. (B) Stimulated by CFP-10. Figure 3. ROC curves and the corresponding AUROC for measurement of frequencies of specific T cells in differentiating ATB and LTBI under stimulation of ESAT-6 or CFP-10. The blue line is drawn with the frequency of IFN-γ-secreting T cells detected by T-SPOT.TB, and the AUC is 0.669 (95%CI, 0.574-0.765). The red line is drawn with combination of the frequencies and proportion of single IFN-γ-'single IL-2-'single TNF-α-'dual IFN-γ/IL-2-'dual IFN-γ/ /TNF-α-'dual IL-2/TNF-α-secreting T cells detected by FluoroSpot, and the AUC is 0.906 (95% CI, 0.856-0.957). Table 1. Diagnostic value of T-SPOT.TB and FluoroSpot for differentiating ATB from LTBI Conclusion Compared with T-SPOT.TB, the IFN-γ/IL-2/TNF-α-Fluorospot assay may be helpful to distinguish ATB from LTBI, and the results need to be verified by large sample prospective cohort study. Disclosures All Authors: No reported disclosures

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Perbedaan Antara Jumlah Sel T Subset Gamma-Delta di Darah Tepi pada Penderita Tuberkulosis dan Orang dengan Latent Tuberculosis Infection

Jurnal Biosains Pascasarjana ◽

10.20473/jbp.v18i2.2016.162-171 ◽

2016 ◽

Vol 18 (2) ◽

pp. 162

Author(s):

Ryzky Widi Atmaja ◽

Jusak Nugraha

Keyword(s):

Mycobacterium Tuberculosis ◽

Heat Shock ◽

Heat Shock Protein ◽

Latent Tuberculosis Infection ◽

Latent Tuberculosis ◽

Tuberculosis Infection ◽

Gamma Delta

Abstrak Latar Belakang. Imunitas memiliki peranan penting untuk melindungi host dari bacilli Mycobacterium tuberculosis (M.tb), bakteri Obligat intraseluler yang menyebabkan Tuberkulosis (TB) dan latent tuberculosis infection (LTBI). Sel T subset gamma-delta (T-γδ) adalah sel-sel potensial tersembunyi yang bermain peran di imunitas innate dan adaptive pada TB. Tetapi, hingga kini perananya di LTBI masih menjadi misteri. Bahan dan Metode. Penelitian dilakukan dengan melibatkan 10 penderita TB serta 10 orang dengan LTBI. Mereka didapatkan dari Rumah Sakit Paru Surabaya melalui suatu persetujuan kelaikan etik dari Universitas Airlangga. Sampel-sampel tersebut akan dihitung jumlah sel T-γδ menggunakan F A C S C a l i b u r. Hasil. Jumlah sel T-γδ meningkat pada TB (10,7%) dan LTBI (15, 4%). Jumlah dari kedua kelompok tersebut melebihi rerata normal di darah tepi (1% - 5%). Kesimpulan. Penigkatan jumlah sel T-γδ pada TB disebabkan melimpahnya kadar IL-12 yang dilepas oleh makrofag selama infeksi. Sementara, peningkatan jumlah sel T-γδ pada LTBI diasumsikan karena banyaknya heat shock protein (HSPs) yang dilepas oleh M.tb di bawah kondisi stres. ...Kata kunci: tuberkulosis, latent tuberculosis infection, Mycobacterium tuberclosis, sel T subset gamma-d e l t a.

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