Improving Assignments for Therapeutic and Prophylactic Treatment Within TB Households. A Potential for Immuno-Diagnosis?

Abstract

Delays in diagnosis and treatment of pulmonary tuberculosis (TB) can lead to more severe disease and increased transmission. Contact investigation among household contacts (HHCs) of TB patients is crucial to ensure optimal outcomes. In the context of a prospective cohort study in Palamaner, Southern India, this study attempted to assess the potential of 27 different soluble immune markers to accurately assign HHCs for appropriate treatment. A multiplex bead assay was applied on QuantiFERON (QFT)-nil supernatants collected from 89 HHCs grouped by longitudinal QFT status; M. tuberculosis (Mtb) infected (QFT positive at baseline and follow-up, n = 30), recent QFT converters (QFT-negative at baseline, n = 27) and converted to QFT-positivity within 6 months of exposure (at follow-up, n = 24) and QFT consistent negatives (n = 32). The 29 TB index cases represented Active TB. Active TB cases and HHCs with Mtb infection produced significantly different levels of both pro-inflammatory (IFNγ, IL17, IL8, IP10, MIP-1α, MIP1β, and VEGF) and anti-inflammatory (IL9 and IL1RA) cytokines. We identified a 4-protein signature (bFGF, IFNγ, IL9, and IP10) that correctly classified HHCs with Mtb infection vs. Active TB with a specificity of 92.6%, suggesting that this 4-protein signature has the potential to assign HHCs for either full-length TB treatment or preventive TB treatment. We further identified a 4-protein signature (bFGF, GCSF, IFNγ, and IL1RA) that differentiated HHCs with Mtb infection from QFT consistent negatives with a specificity of 62.5%, but not satisfactory to safely assign HHCs to no preventive TB treatment. QFT conversion, reflecting new Mtb infection, induced an elevated median concentration in nearly two-thirds (19/27) of the analyzed soluble markers compared to the levels measured at baseline. Validation in other studies is warranted in order to establish the potential of the immune biosignatures for optimized TB case detection and assignment to therapeutic and preventive treatment of Mtb infected individuals.

Keywords: Mtb infection; active TB; cytokine and chemokines; preventive therapy; protein signature; soluble protein markers.

Figure 1

Study flow chart.

Figure 2

Kruskal–Wallis test with Dunn’s post correction was applied. Scatter-plot graph depicting median cytokine concentrations (pg ml⁻¹) from the QFT-GIT supernatants (nil tube) by the Multiplex bio-plex assay. Nil levels (pg ml⁻¹) of IFNγ, IL9, IP10, IL-17, VEGF, GCSF, IL1RA, IL8, MIP1α, and MIP1β in patients with active TB (ATB; n = 29), Mtb infected (n = 30), recent QFT converters (n = 27), and QFT consistent negatives (n = 32).

Figure 3

Wilcoxon matched paired test was applied. Symbols and lines graph depicting individual changes in cytokines/chemokines concentrations (pg ml⁻¹) from recent QFT converters in baseline samples (n = 27) to recent QFT converters in the follow-up sample (n = 24).

Figure 4

ROC curves for protein signature that distinguishes (A) Active TB (ATB; n = 29) from Mtb infected (n = 30), (B) ATB (n = 29) from recent QFT converters (n = 27), (C) ATB (n = 29) from QFT consistent negatives (n = 32), and (D) Mtb infected (n = 30) from QFT consistent negatives (n = 32).

Figure 5

ROC curves for signature that distinguishes (A) Active TB (n = 29) from Household contacts with Mtb infection (n = 30), (B) Household contacts with Mtb infection (n = 30) from QFT consistent negatives (n = 32).