Ji-Sook Lee; Ji-Yeon Lee; Hong-Hee Choi; Ji Woong Son; Ki-Hye Kim; Tae-Hyun Paik; Eun-Kyeong Jo

doi:10.4167/jbv.2007.37.3.137

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Lee, Lee, Choi, Son, Kim, Paik, and Jo: Elevated Levels of Interferon-inducible Protein-10 (IP)-10/CXCL10, but not of Interferon-γ, in Patients with Pulmonary Tuberculosis

Original Article

J Bacteriol Virol 2007;37(3):137-146.

Published online: 18 February 2007

DOI: https://doi.org/10.4167/jbv.2007.37.3.137

Elevated Levels of Interferon-inducible Protein-10 (IP)-10/CXCL10, but not of Interferon-γ, in Patients with Pulmonary Tuberculosis

Ji-Sook Lee¹, Ji-Yeon Lee¹, Hong-Hee Choi¹, Ji Woong Son², Ki-Hye Kim¹, Tae-Hyun Paik³, Eun-Kyeong Jo^1,^4,^∗

¹Department of Microbiology, College of Medicine, Chungnam National University, Daejeon 301-747, Republic of Korea

²Department of Internal Medicine, Konyang University, Daejeon 302-718, Republic of Korea

³Microbiology, College of Medicine, Konyang University, Daejeon 302-718, Republic of Korea

⁴Infection Signaling Network Research Center, College of Medicine, Chungnam National University, Daejeon 301-747, Republic of Korea

^∗Corresponding author: Prof. Dr. Eun-Kyeong Jo, Department of Microbiology, College of Medicine, Chungnam National University, 6 Munhwa-dong, Jung-ku, Daejeon 301-747, Republic of Korea. Phone: +82-42-580-8243, Fax: +82-42-585-3686 e-mail: hayoungj@cnu.ac.kr

^∗∗ This work was supported by a grant of the Korea Science & Engineering Foundation through the Infection Signaling Network Research Center (R13-2007-020-01000–0) at Chungnam National University. The authors declare that they have no competing financial interests.

Received 23 August 2007 Accepted 19 September 2007

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Mycobacterial strains are potent inducers of cytokines/chemokines by mononuclear phagocytes, which constitute an important cellular component of the first line of defense in the innate immune system. Interferon (IFN)-γ-inducible protein (IP-10 or CXCL10) is a potent chemoattractant; however, little is known about the IP-10 profiles attributable to the Th1 regulation associated with active tuberculosis (TB). In this study, we investigated the production of IP-10, interleukin (IL)-12 p40, and IFN-γ by the peripheral blood mononuclear cells (PBMCs) of patients with active pulmonary TB in response to in vitro stimulation with Triton X-100 soluble proteins (TSPs) or the 30-kDa antigen. The TSP antigens used in the present study were isolated and purified from Mycobacterium tuberculosis H37Rv (virulent strain), M. tuberculosis H37Ra (avirulent strain), and Mycobacterium bovis BCG. The results were compared with those obtained for healthy tuberculin reactors (HTRs). Concordant with earlier studies, IFN-γ production was significantly depressed in the PBMCs from TB patients compared with those in the HTR group. However, the IP-10 levels in the PBMCs from TB patients were significantly elevated 18 h after stimulation compared to those in the PBMCs from HTRs. IP-10 release was correlated in a significant manner with the release of IFN-γ in the HTRs, but this was not the case for the TB patients. Collectively, these data suggest that TB patients show altered regulation of Th1-driving cytokine and chemokine production in response to a variety of mycobacterial antigens.

Keywords: Tuberculosis, Mycobacterium tuberculosis, Interferon-γ-inducible protein, Interferon-γ, 30-kDa antigen, TSP antigen

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Figure 1.

SDS-PAGE analysis of TSP Ags and 30-kDa Ag of M. tuberculosis with Coomassie blue staining. The TSP-Rv, TSP-Ra, and TSP-BCG Ags were isolated from M. tuberculosis H37Rv, M. tuberculosis H37Ra, and M. bovis BCG, respectively, by overnight solubilization with Triton X-100, 20% to 80% ammonium sulfate precipitation, three rounds of phase partitioning with Triton X-114, and three rounds of 0% to 80% ammonium sulfate precipitation. The 30-kDa Ag was purified from unheated concentrated culture filtrate of M. tuberculosis H37Rv by 0% to 40% ammonium sulfate precipitation, anion-exchange chromatography, and hydrophobic interaction chromatography. A, TSP-Rv; B, TSP-Ra; C, TSP-BCG; D, 30-kDa Ag.

Figure 2.

IFN-γ (A) and IL-12p40 (B) production by PBMCs from TB patients in response to TSP-Rv, TSP-Ra, TSP-BCG, and 30-kDa Ag. IFN-γ and IL-12p40 production by PBMCs from HTRs and TB patients were determined after in vitro stimulation with TSP-Rv, TSP-Ra, TSP-BCG, and 30-kDa Ag (1 μg/ml for each Ag). The supernatants were harvested at 18 h (for IL-12p40) or 96 h (for IFN- γ) for the assessment of cytokines by ELISA. The values are shown as the mean ± SD of the supernatant samples. ^∗p<0.05 and ^∗∗p<0.01, as compared with the HTRs (Student's t-test). Us, unstimulated.

Figure 3.

IP-10 production by PBMCs from TB patients in response to TSP-Rv, TSP-Ra, TSP-BCG, and 30-kDa Ag. The IP-10 production levels by PBMCs from HTRs and TB patients were determined after in vitro stimulation with TSP-Rv, TSP-Ra, TSP-BCG, and 30-kDa Ag (1 μg/ml for each Ag). The supernatants were harvested at 18 h (A) and 96 h (B), and the IP-10 protein levels were measured by ELISA. The values are shown as the mean ± SD of supernatant samples. ^∗p<0.05 and ^∗∗p<0.01, as compared with the HTRs (Student's t-test). Us, unstimulated.

Figure 4.

Linear regression analysis of IFN-γ and IP-10 production in TB patients and HTRs. The levels of IP-10 and IFN-γ release were assayed by ELISA after stimulation with the TSP-Rv and 30-kDa Ags for 96 h. A significant correlation was found between IP-10 and IFN-γ production after in vitro stimulation with 30-kDa Ag or TSP-Rv Ag in HTRs (n=10, r=0.84, and p<0.05 for the 30-kDa Ag; n=9, r=0.74, and p<0.05 for TSP Rv), but not found in TB patients (n=10, r=0.19, and NS for the 30-kDa Ag; n=10, r=0.000, and NS for TSP Rv). NS, not significant.

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