Immunological aspectsProfiling the human immune response to Mycobacterium tuberculosis by human cytokine array
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
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