J Bacteriol Virol.  2007 Sep;37(3):137-146. 10.4167/jbv.2007.37.3.137.

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

• Lee JS
• Lee JY
• Choi HH
• Son JW
• Kim KH
• Paik TH
• Jo EK

• KMID: 1513568
• DOI: http://doi.org/10.4167/jbv.2007.37.3.137

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)-gamma-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-gamma 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-gamma 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-gamma 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.

Keyword

Tuberculosis; Mycobacterium tuberculosis; Interferon-gamma-inducible protein; Interferon-gamma; 30-kDa antigen; TSP antigen

MeSH Terms

Humans
Immune System
Interferon-gamma*
Interferons
Interleukins
Mycobacterium bovis
Mycobacterium tuberculosis
Octoxynol
Phagocytes
Tuberculin
Tuberculosis
Tuberculosis, Pulmonary*
Interferon-gamma
Interferons
Interleukins
Octoxynol
Tuberculin

Figure

• 

  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.

Reference

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