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Dominant TNF-α+ Mycobacterium tuberculosis–specific CD4+ T cell responses discriminate between latent infection and active disease

Nature Medicine volume 17, pages 372–376 (2011)Cite this article

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Abstract

Rapid diagnosis of active Mycobacterium tuberculosis (Mtb) infection remains a clinical and laboratory challenge. We have analyzed the cytokine profile (interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α) and interleukin-2 (IL-2)) of Mtb-specific T cells by polychromatic flow cytometry. We studied Mtb-specific CD4+ T cell responses in subjects with latent Mtb infection and active tuberculosis disease. The results showed substantial increase in the proportion of single-positive TNF-α Mtb-specific CD4+ T cells in subjects with active disease, and this parameter was the strongest predictor of diagnosis of active disease versus latent infection. We validated the use of this parameter in a cohort of 101 subjects with tuberculosis diagnosis unknown to the investigator. The sensitivity and specificity of the flow cytometry–based assay were 67% and 92%, respectively, the positive predictive value was 80% and the negative predictive value was 92.4%. Therefore, the proportion of single-positive TNF-α Mtb-specific CD4+ T cells is a new tool for the rapid diagnosis of active tuberculosis disease.

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Figure 1: Quantitative and qualitative analysis of Mtb-specific T cell responses in the test cohort.
Figure 2: Analysis of Mtb-specific T cell responses in the validation cohort after unblinding of the clinical status.
Figure 3: Percentages of CFP-10– or ESAT-6–specific single-positive TNF-α–producing CD4+ T cells of the 94 subjects from the validation cohort with concordant responses against CFP-10 and ESAT-6.
Figure 4: Longitudinal analysis of the functional profile of Mtb-specific CD4+ T cells from five subjects analyzed during untreated active tuberculosis disease and then after tuberculosis treatment.

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Acknowledgements

This research was partially conducted as part of the Vaccine Immune Monitoring Consortium under the Collaboration for AIDS Vaccine Discovery with support from the Bill & Melinda Gates Foundation. Furthermore, we thank N. Rettby, D. Bonnet and K. Ellefsen Lavoie for logistic coordination. We also thank many additional members of the South African Tuberculosis Vaccine Initiative team who helped with enrollment and evaluation of participants and, finally, the participants themselves.

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Authors and Affiliations

  1. Division of Immunology and Allergy, Centre Hospitalier Universitaire Vaudois, University of Lausanne, Lausanne, Switzerland

    Alexandre Harari, Virginie Rozot, Felicitas Bellutti Enders, Matthieu Perreau, Pierre-Alexandre Bart & Giuseppe Pantaleo

  2. Swiss Vaccine Research Institute, Lausanne, Switzerland

    Alexandre Harari & Giuseppe Pantaleo

  3. Division of Pneumology, Centre Hospitalier Universitaire Vaudois, University of Lausanne, Lausanne, Switzerland

    Jesica Mazza Stalder & Laurent P Nicod

  4. Division of Infectious Diseases, Centre Hospitalier Universitaire Vaudois, University of Lausanne, Lausanne, Switzerland

    Matthias Cavassini & Thierry Calandra

  5. Division of Occupational Medicine, Centre Hospitalier Universitaire Vaudois, University of Lausanne, Lausanne, Switzerland

    Catherine Lazor Blanchet

  6. Institute of Microbiology, Centre Hospitalier Universitaire Vaudois, University of Lausanne, Lausanne, Switzerland

    Katia Jaton

  7. Center of Clinical Epidemiology, Centre Hospitalier Universitaire Vaudois, University of Lausanne, Lausanne, Switzerland

    Mohamed Faouzi

  8. South African Tuberculosis Vaccine Initiative, University of Cape Town, Cape Town, South Africa

    Cheryl L Day & Willem A Hanekom

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  1. Alexandre Harari
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Contributions

A.H. designed the study, performed the analyses and wrote the manuscript; V.R., F.B.E. and M.P. generated data and performed analyses; J.M.S., L.P.N., M.C., T.C., C.L.B., C.L.D. and W.A.H. recruited study participants; K.J. performed analyses; M.F. performed the statistical analyses; P.-A.B. contributed to the design of the study, performed analyses and wrote the manuscript; G.P. designed the study, supervised the analyses and wrote the paper. All authors have read and approved the final manuscript.

Corresponding author

Correspondence to Giuseppe Pantaleo.

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The authors declare no competing financial interests.

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Supplementary Tables 1 and 2 and Supplementary Figures 1–7 (PDF 389 kb)

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Harari, A., Rozot, V., Enders, F. et al. Dominant TNF-α+ Mycobacterium tuberculosis–specific CD4+ T cell responses discriminate between latent infection and active disease. Nat Med 17, 372–376 (2011). https://doi.org/10.1038/nm.2299

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