Antigen-Specific Cytokine and Chemokine Gene Expression for Diagnosing Latent and Active Tuberculosis

Workneh Korma; Adane Mihret; Yunhee Chang; Azeb Tarekegn; Metasebiya Tegegn; Adem Tuha; Dasom Hwang; Mesfin Asefa; Mahlet O Hasen; Seoyoung Kim; Tesfaye S Tesema; Hyeyoung Lee

doi:10.3390/diagnostics10090716

Antigen-Specific Cytokine and Chemokine Gene Expression for Diagnosing Latent and Active Tuberculosis

Diagnostics (Basel). 2020 Sep 18;10(9):716. doi: 10.3390/diagnostics10090716.

Authors

Affiliations

¹ Molecular Diagnostic Laboratory, Department of Biomedical Laboratory Sciences, Yonsei University, Wonju 26493, Korea.
² Institute of Biotechnology, Addis Ababa University, Addis Ababa, P.O. Box 1176, Ethiopia.
³ Armauer Hansen Research Institute, Addis Ababa, P.O Box 1005, Ethiopia.
⁴ St. Paul's Hospital Millennium Medical College, Department of pathology, Addis Ababa, P.O. Box 1271, Ethiopia.

Abstract

Tuberculosis infection exhibits different forms, namely, pulmonary, extrapulmonary, and latent. Here, diagnostic markers based on the gene expression of cytokines and chemokines for differentiating between tuberculosis infection state(s) were identified. Gene expression of seven cytokines (Interferon gamma (IFN-γ), Interferon gamma-induced protein 10 (IP-10), Interleukin-2 receptor (IL-2R), C-X-C Motif Chemokine Ligand 9 (CXCL-9), Interleukin 10 (IL-10), Interleukin 4 (IL-4), and Tumor Necrosis Factor alpha (TNF-α)) in response to tuberculosis antigen was analyzed using real-time polymerase reaction. The sensitivity and specificity of relative quantification (2^^-ΔΔCt) of mRNA expression were analyzed by constructing receiver operating characteristic curves and measuring the area under the curve (AUC) values. Combinations of cytokines were analyzed using the R statistical software package. IFN-γ, IP-10, IL2R, and CXCL-9 showed high expression in latent and active tuberculosis patients (p = 0.001), with a decrease in IL10 expression, and no statistical difference in IL-4 levels among all the groups (p = 0.999). IL-10 differentiated pulmonary tuberculosis patients from latent cases with an AUC of 0.731. IL10 combined with CXCL-9 distinguished pulmonary tuberculosis patients from extrapulmonary cases with a sensitivity, specificity, and accuracy of 85.7%, 73.9%, and 81.0%, respectively. IL-10 together with IP-10 and IL-4 differentiated pulmonary tuberculosis from latent cases with a sensitivity and specificity of 77.1% and 88.1%, respectively. Decision tree analysis demonstrated that IFN-γ IL-2R, and IL-4 can diagnose tuberculosis infection with a sensitivity, specificity, and accuracy of 89.7%, 96.1%, and 92.7%, respectively. A combination of gene expression of cytokines and chemokines might serve as an effective marker to differentiate tuberculosis infection state(s).

Keywords: RT-PCR; Tuberculosis diagnosis; chemokines; cytokine and chemokine biomarker; cytokines; diagnostics; extrapulmonary tuberculosis; gene expression; tuberculosis biomarker.

Abstract

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