Differed IL-1 Beta Response between Active TB and LTBI Cases by Ex Vivo Stimulation of Human Monocyte-Derived Macrophage with TB-Specific Antigen

Abstract

Background. The difference of macrophage-specific interleukin-1 beta (IL-1b) response between latent tuberculosis infection (LTBI) and active tuberculosis (TB) remains less studied. Method. We performed this prospective study and recruited active TB patients, contacts with LTBI, and uninfected contacts. The gene and protein expression of human monocyte-derived macrophage (hMDM) after ex vivo stimulation by early secretory antigenic target-6KD (ESAT-6) and tuberculin purified protein derivatives (PPD) was studied by real-time PCR and flow cytometry. The effect of caspase-1 inhibitor was also studied. Result. The IL-1b gene expression after 6 hr ESAT-6 1 μg/ml stimulation was different among active TB patients (n=12), LTBI cases (n=12), and uninfected contacts (n=23) (log fold change: 0.98±1.26 vs. 2.20±0.96 vs. 2.20±0.96, P=0.013). The IL-1b gene expression at 24 hours was higher than that at 6 hours in LTBI cases (n=4) and uninfected contacts (n=6). After 24 hr ESAT-6 1 μg/ml stimulation, the percentage of IL-1b-expressed hMDM was borderline lower in the active TB patients (n=9) than in the LTBI cases (n=10) (14.0±11.2% vs. 31.6±22.5%, P=0.065). Compared with ESAT-6 1 μg/ml stimulation but without the addition of caspase-1 inhibitor (CasI) (55.6±16.3%), the percentage of IL-1b-positive hMDMs decreased after addition of CasI (50 μg/ml CasI: 49.8±18.2%, P=0.078; 100 μg/ml CasI: 46.6±20.8%, P=0.030; 150 μg/ml CasI: 33.7±15.5%, P=0.016). Conclusions. This study revealed that macrophage-specific IL-1b response differed among different stages of Mycobacterium tuberculosis infection. The role of IL-1b and inflammasome in the process of LTBI progressing to active TB warrants further investigation.