Comparative Proteomics of Activated THP-1 Cells Infected with Mycobacterium tuberculosis Identifies Putative Clearance Biomarkers for Tuberculosis Treatment

Abstract

Biomarkers for determining clearance of Mycobacterium tuberculosis (Mtb) infection during anti-tuberculosis therapy or following exposure could facilitate enhanced monitoring and treatment. We screened for biomarkers indicating clearance of Mtb infection in vitro. A comparative proteomic analysis was performed using GeLC MSI/MS. Intracellular and secreted proteomes from activated THP-1 cells infected with the Mtb H37Rv strain (MOI = 1) and treated with isoniazid and rifampicin for 1 day (infection stage) and 5 days (clearance stage) were analyzed. Host proteins associated with early infection (n = 82), clearance (n = 121), sustained in both conditions (n = 34) and suppressed by infection (n = 46) were elucidated. Of the potential clearance markers, SSFA2 and CAECAM18 showed the highest and lowest protein intensities, respectively. A western blot of CAECAM18 validated the LC MS/MS result. For three clearance markers (SSFA2, PARP14 and PSME4), in vivo clinical validation was concordantly reported in previous patient cohorts. A network analysis revealed that clearance markers were enriched amongst four protein interaction networks centered on: (i) CD44/CCND1, (ii) IFN-β1/NF-κB, (iii) TP53/TGF-β and (iv) IFN-γ/CCL2. After infection, proteins associated with proliferation, and recruitment of immune cells appeared to be enriched possibly reflecting recruitment of defense mechanisms. Counteracting proteins (CASP3 vs. Akt and NF-κB vs. TP53) associated with apoptosis regulation and its networks were enriched among the early and sustained infection biomarkers, indicating host-pathogen competition. The BRCA1/2 network was suppressed during infection, suggesting that cell proliferation suppression is a feature of Mtb survival. Our study provides insights into the mechanisms of host-Mtb interaction by comparing the stages of infection clearance. The identified clearance biomarkers may be useful in monitoring tuberculosis treatment.

Proteomic analysis of infected primary human leucocytes revealed PSTK as potential treatment-monitoring marker for active and latent tuberculosis

“…The discovery of Mtb clearance markers that can be used as TB treatment monitoring markers is difficult in part because of the difficulty of confirming clearance of Mtb from the tissues of patients. Previously, we successfully demonstrated that THP-1 cells infected with Mtb strain H37Rv could be treated with anti-TB drugs leading to confirmed in-vitro clearance of bacilli [13]. With a simple cell-line model, we demonstrated that SSFA2, NCOR2, MCM2, RANBP1, RPTN, ELFN1, RFC2, SETX and CEACAM18 were potential clearance biomarkers [13].…”

“…Previously, we successfully demonstrated that THP-1 cells infected with Mtb strain H37Rv could be treated with anti-TB drugs leading to confirmed in-vitro clearance of bacilli [13]. With a simple cell-line model, we demonstrated that SSFA2, NCOR2, MCM2, RANBP1, RPTN, ELFN1, RFC2, SETX and CEACAM18 were potential clearance biomarkers [13]. In the current study, we used a more complex model with stringent criteria to select candidate clearance biomarkers that could be used clinically.…”

“…The activated cells were infected with four Mtb strains (EA, IO, EuA and H37Rv) and incubated for 4 hours. Then, the infected cells were treated with a combination of isoniazid (INH) and rifampicin (RIF) with 3: 9 μg/ml of INH:RIF (optimized concentrations according to a previous study [13] for intracellular bacillary killing within three days). Medium containing the drugs was replaced in cell cultures every 24 hours for up to five days.…”

“…Previously, our group reported a preliminary study exploring potential clearance markers using a monocytic cell line (THP-1 cells) [13]. However, testing using a wider range of Mtb lineages and other bacteria as controls is required, as well as clinical sample validation.…”

Breast and Cervix Uteri: Rare Locations for Mycobacterium Tuberculosis Infections and Complications-Cases Report and Literature Review

“…The breast was considered an organ that offers resistance to the survival and multiplication of the tubercle bacillus [24], like other organs such as spleen, skeletal muscle or cervix uteri, and this property is one of the reasons for the uncommon diagnosis of breast TB, and in countries like India it is supposed that the breast TB is confused to carcinoma [25], or to a pyogenic breast abscess [26], or to other diseases [27], tubercular mastitis being a "great masquerader" [28]. In contrast to this quality, when breast is infected with MbT, it was proved that BRCA 1/2 network is suppressed during infection suggesting that breast cell proliferation suppression is a feature of Koch bacilli survival [29].…”