Elsevier

Tuberculosis

Volume 97, March 2016, Pages 108-117
Tuberculosis

Immunological aspects
Profiling the human immune response to Mycobacterium tuberculosis by human cytokine array

https://doi.org/10.1016/j.tube.2015.12.007Get rights and content

Summary

Object

Tuberculosis (TB) continues to be one of the most serious infectious diseases in the world, however, no effective biomarkers can be used for rapid screening of latent tuberculosis infection (LTBI) and active TB. In this study, serum cytokines were screened and tested as potential biomarker for TB diagnosis.

Method

Cytokine array was used to track the cytokine profile and its dynamic change after TB infection. The different expressions of cytokines were confirmed by ELISA assay. ROC curve analyses were used to evaluate the efficacy of a cytokine or cytokine combination for diagnosis.

Results

Eotaxin-2, ICAM-1, MCSF, IL-12p70, and IL-11 were significantly higher in the LTBI individuals. I-309, MIG, Eotaxin-2, IL-8, ICAM-1, IL-6sR, and Eotaxin were significantly higher in active TB patients. ROC curve analyses gave AUCs of 0.843, 0.898, and 0.888 for I-309, MIG, and IL-8, respectively, and 0.894 for the combination panel in active TB diagnosis. IFN-γ/IL-4 and IL-2/TNF-α ratios exhibit dynamic changes in the healthy control and LTBI to different stages of active TB.

Conclusions

Serum cytokines, including I-309 and MIG, IL-8, Extoxin-2, ICAM-1 and combinations of cytokines, including IFN-γ/IL-4 and IL-2/TNF-α, can be used as serum biomarkers for LTBI and active TB screening, thus indicating prospective clinical applications.

Introduction

Tuberculosis (TB) remains a public health threat and is the second leading cause of death from an infectious disease worldwide [1]. Considerable effort has been directed toward controlling tuberculosis. Recent data show there were 8.6 million new TB cases and 1.3 million TB deaths in 2012 [2]. Approximately 32% of the world population is infected with Mycobacterium tuberculosis (MTB) and remains asymptomatic [2]. Of these latently infected individuals, an estimated 5–10% will develop into active tuberculosis in their lifetime [3]. Although these LTBI are not infectious, identification of LTBI is important for immediate intervention as it is the most effective way to control TB.

At present, effective means of TB and LTBI diagnosis are still lacking. Although the tuberculin skin test (TST) is rapid and inexpensive, it is vulnerable to the influence of BCG vaccination [4]. In China, more than 800 million people have received the BCG vaccine; thus, TST in China was proven ineffective for diagnosis [5], [6]. The IFN-γ release assay (IGRA) is not affected by BCG vaccination, but the process involved in the assay is complicated and costly, making it unsuitable for mass screening [7]. The recently developed POCT product is supposedly the best product for mass screening. However, the product was not sensitive and specific enough in clinical applications [8]. Therefore, TB diagnosis and LTB screening required new technology and new biomarkers.

The outcome of natural infections with pathogenic mycobacteria can range from early asymptomatic clearance through latent infection to clinical disease. Different host- and pathogen-specific factors have been implicated in the outcome of these infections. However, the interaction of mycobacteria with the innate and acquired components of the immune system clearly plays a central role in the infections. The cytokine response induced by mycobacteria has a critical effect on the development of the disease by limiting bacterial growth and regulating inflammation [9]. Therefore, understanding the behavior of different cytokines during the different stages of pathogenesis is important. Several cytokines have been implicated in the pathogenesis and control of MTB infection. These cytokines act as messengers that help integrate immune system components in the fight against invading pathogens. Proinflammatory cytokines released by polymorphonuclear neutrophils contribute to the recruitment of leukocytes at the infection site and amplify the immune response [10]. Related immune factors, such as IFN-γ [11] and IP-10 [12], are highly expressed in the serum of patients with active TB. Moreover, IL-2 is significantly up-regulated in LTBIs [13]; thus, it may be used as a marker for LTBI screening. Determining the pathways by which specific cytokines modulate the immune response to infection may enable the development of effective interventions.

In this study, we described the changes in cytokine profile of patients infected with TB and the dynamic cytokine response to anti-TB chemotherapy by using a high-throughput protein microarray platform. In order to identify a good indicator for LTBI and active TB diagnosis, we determine the cytokine profile response in TB infection and chemotherapy with different pathogenesis.

Section snippets

Ethics statement and sample selection

This study was reviewed and approved by the local ethics committee (Guangdong Center for Tuberculosis Control). Written informed consent was obtained from participants before they were enrolled into the study, and written informed consent was obtained from the guardians on behalf of the children enrolled.

In this study, 294 participants aged 13–65 years were enrolled from Guangzhou, Shenzhen, and Foshan cities of Guangdong province of China. In human cytokines microarray assay, 96 peripheral

Serum cytokine profile and its dynamic change response to TB infection and anti-TB chemotherapy

To screen the serum cytokine biomarkers for LTBI and active TB, we employed antibody array technology to determine the expression profiles of 40 human cytokines in peripheral serum samples of healthy controls (N), LTB infections (I), active TB patients (Pa), and active TB patients under chemotherapy at different time points (Pb: 2 months; Pc: 6 months). The standard period of TB treatment is six months, including two months of intensive chemotherapy followed by four months of consolidated

Discussion

In this study, we systematically compared the cytokine expression profiles of healthy controls, LTBI, and active TB patients to identify the cytokine biomarkers for TB infection and anti-TB chemotherapy. The changes in cytokine expression pattern were also continuously observed during anti-TB chemotherapy. The cytokine expression profile in the serum of humans with LTBI using high-throughput array, which is different from the stimulation of the whole blood or monocytes from LTBI with

Acknowledgments

This study was funded by grants from the National Science and Technology Key Project for Major Infectious Diseases (2012ZX10004903, 2013ZX10003001, and 2014ZX10003002), Guangdong Natural Science Foundation (WSTJJ20120319130623198210011211), Science and Technology of Guangdong Province (2013B051000055). We thank Mr. Wushan Yin (CTK Biotechnology) who kindly provided the TB specific antigens Ag14-16KDa, Ag32KDa, Ag 38KDa and Ag85B. We also thank Mr. Qingjin Wu for his kind assistance with the

References (44)

  • V. Boggaram et al.

    Early secreted antigenic target of 6 kDa (ESAT-6) protein of Mycobacterium tuberculosis induces interleukin-8 (IL-8) expression in lung epithelial cells via protein kinase signaling and reactive oxygen species

    J Biol Chem

    (2013)
  • A. Zumla et al.

    Tuberculosis

    N Engl J Med

    (2013)
  • P. Glaziou et al.

    Global epidemiology of tuberculosis preface

    Semin Respir Crit Care Med

    (2013)
  • E. Vynnycky et al.

    Lifetime risks, incubation period, and serial interval of tuberculosis

    Am J Epidemiol

    (2000)
  • S. Burl et al.

    The tuberculin skin test (TST) is affected by recent BCG vaccination but not by exposure to non-tuberculosis mycobacteria (NTM) during early life

    PLoS One

    (2010)
  • R. Turner et al.

    No proof that BCG protects against infection with Mycobacterium tuberculosis

    BMJ

    (2014)
  • C. Herzmann et al.

    Early BCG vaccination is unrelated to pulmonary immunity against Mycobacterium tuberculosis in adults

    Eur Respir J

    (2014)
  • K. Kruczak et al.

    Comparison of IGRA tests and TST in the diagnosis of latent tuberculosis infection and predicting tuberculosis in risk groups in Krakow, Poland

    Scand J Infect Dis

    (2014)
  • E. Torrado et al.

    Cytokines in the balance of protection and pathology during mycobacterial infections

    Adv Exp Med Biol

    (2013)
  • E. Borgstrom et al.

    Immune responses to ESAT-6 and CFP-10 by FASCIA and multiplex technology for diagnosis of M. tuberculosis infection; IP-10 is a promising marker

    PLoS One

    (2012)
  • Y.Y. Chen et al.

    The pattern of cytokine production in vitro induced by ancient and modern Beijing Mycobacterium tuberculosis strains

    PLoS One

    (2014)
  • P.N. Essone et al.

    Host cytokine responses induced after overnight stimulation with novel M. tuberculosis infection phase-dependent antigens show promise as diagnostic candidates for TB disease

    PLoS One

    (2014)
  • Cited by (0)

    1

    Corresponding first author.

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