Letter | Published:

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

Nature Medicine volume 17, pages 372376 (2011) | Download Citation


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.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.


  1. 1.

    How can immunology contribute to the control of tuberculosis? Nat. Rev. Immunol. 1, 20–30 (2001).

  2. 2.

    & Immunology of tuberculosis. Annu. Rev. Immunol. 19, 93–129 (2001).

  3. 3.

    et al. Enhanced contact tracing and spatial tracking of Mycobacterium tuberculosis infection by enumeration of antigen-specific T cells. Lancet 357, 2017–2021 (2001).

  4. 4.

    et al. Comparison of T cell–based assay with tuberculin skin test for diagnosis of Mycobacterium tuberculosis infection in a school tuberculosis outbreak. Lancet 361, 1168–1173 (2003).

  5. 5.

    , , , & Sensitivity of a new commercial enzyme-linked immunospot assay (T SPOT-TB) for diagnosis of tuberculosis in clinical practice. Eur. J. Clin. Microbiol. Infect. Dis. 24, 529–536 (2005).

  6. 6.

    , & Clinical practice. Latent tuberculosis infection. N. Engl. J. Med. 347, 1860–1866 (2002).

  7. 7.

    et al. HIV nonprogressors preferentially maintain highly functional HIV-specific CD8+ T cells. Blood 107, 4781–4789 (2006).

  8. 8.

    et al. Functional signatures of protective antiviral T cell immunity in human virus infections. Immunol. Rev. 211, 236–254 (2006).

  9. 9.

    & Functional signatures in antiviral T cell immunity for monitoring virus-associated diseases. Nat. Rev. Immunol. 6, 417–423 (2006).

  10. 10.

    & Correlates of immune protection in HIV-1 infection: what we know, what we don't know, what we should know. Nat. Med. 10, 806–810 (2004).

  11. 11.

    et al. Detection of polyfunctional Mycobacterium tuberculosis–specific T cells and association with viral load in HIV-1–infected persons. J. Infect. Dis. 197, 990–999 (2008).

  12. 12.

    , , , & Pattern and diversity of cytokine production differentiates between Mycobacterium tuberculosis infection and disease. Eur. J. Immunol. 39, 723–729 (2009).

  13. 13.

    et al. Tumor necrosis factor-α is required in the protective immune response against Mycobacterium tuberculosis in mice. Immunity 2, 561–572 (1995).

  14. 14.

    & Anti-TNF α therapy of rheumatoid arthritis: what have we learned? Annu. Rev. Immunol. 19, 163–196 (2001).

  15. 15.

    et al. Infliximab (chimeric anti-tumour necrosis factor α monoclonal antibody) versus placebo in rheumatoid arthritis patients receiving concomitant methotrexate: a randomised phase III trial. ATTRACT Study Group. Lancet 354, 1932–1939 (1999).

  16. 16.

    et al. An interferon-inducible neutrophil-driven blood transcriptional signature in human tuberculosis. Nature 466, 973–977 (2010).

  17. 17.

    , & Intracellular cytokine optimization and standard operating procedure. Nat. Protoc. 1, 1507–1516 (2006).

  18. 18.

    et al. An HIV-1 clade C DNA prime, NYVAC boost vaccine regimen induces reliable, polyfunctional and long-lasting T cell responses. J. Exp. Med. 205, 63–77 (2008).

  19. 19.

    et al. HIV-1-specific IFN-γ/IL-2–secreting CD8 T cells support CD4-independent proliferation of HIV-1–specific CD8 T cells. Proc. Natl. Acad. Sci. USA 102, 7239–7244 (2005).

  20. 20.

    , , & Selection and interpretation of diagnostic tests and procedures. Principles and applications. Ann. Intern. Med. 94, 557–592 (1981).

  21. 21.

    Basic principles of ROC analysis. Semin. Nucl. Med. 8, 283–298 (1978).

  22. 22.

    & Receiver-operating characteristic (ROC) plots: a fundamental evaluation tool in clinical medicine. Clin. Chem. 39, 561–577 (1993).

Download references


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.

Author information


  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


  1. Search for Alexandre Harari in:

  2. Search for Virginie Rozot in:

  3. Search for Felicitas Bellutti Enders in:

  4. Search for Matthieu Perreau in:

  5. Search for Jesica Mazza Stalder in:

  6. Search for Laurent P Nicod in:

  7. Search for Matthias Cavassini in:

  8. Search for Thierry Calandra in:

  9. Search for Catherine Lazor Blanchet in:

  10. Search for Katia Jaton in:

  11. Search for Mohamed Faouzi in:

  12. Search for Cheryl L Day in:

  13. Search for Willem A Hanekom in:

  14. Search for Pierre-Alexandre Bart in:

  15. Search for Giuseppe Pantaleo in:


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.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Giuseppe Pantaleo.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Tables 1 and 2 and Supplementary Figures 1–7

About this article

Publication history






Further reading