PT  - JOURNAL ARTICLE
AU  - Darragh Duffy
AU  - Elisa Nemes
AU  - Alba Llibre
AU  - Vincent Rouilly
AU  - Elizabeth Filander
AU  - Hadn Africa
AU  - Simbarashe Mabwe
AU  - Lungisa Jaxa
AU  - Bruno Charbit
AU  - Munyaradzi Musvosvi
AU  - Humphrey Mulenga
AU  - the Milieu Intérieur Consortium
AU  - Stephanie Thomas
AU  - Mark Hatherill
AU  - Nicole Bilek
AU  - Thomas J Scriba
AU  - Matthew L Albert
TI  - Immune profiling in M. tuberculosis infection enables stratification of patients with active disease
AID  - 10.1101/581298
DP  - 2019 Jan 01
TA  - bioRxiv
PG  - 581298
4099  - http://biorxiv.org/content/early/2019/03/18/581298.short
4100  - http://biorxiv.org/content/early/2019/03/18/581298.full
AB  - Tuberculosis (TB) is caused by Mycobacterium tuberculosis (Mtb) infection and is a major public health problem with an estimated 1.7 billion persons infected worldwide. Clinical challenges in TB include the lack of a blood-based test for active disease, and the absence of prognostic biomarkers for early treatment response. Current blood based tests, such as QuantiFERON-TB Gold (QFT), are based on an IFNγ readout following Mtb antigen stimulation. However, they do not distinguish active TB disease from asymptomatic Mtb infection. We hypothesized that the use of TruCulture, an improved immunomonitoring method for whole blood collection and immune stimulation, could improve the discrimination of active disease from latent Mtb infection. To test our hypothesis, we stimulated whole blood from active TB patients (before and after successful treatment), comparing them to asymptomatic latently infected individuals. Mtb-specific antigens (ESAT-6, CFP-10, TB7.7) and live bacillus Calmette-Guerin (BCG) were used for TruCulture stimulation conditions, with direct comparison to QFT. Protein analyses were performed on the culture supernatants using ELISA and Luminex multi-analyte profiling. TruCulture showed an ability to discriminate active TB cases from latent controls (p < 0.0001, AUC = 0.81, 95% CI: 0.69-0.93) as compared to QFT (p = 0.47 AUC = 0.56, 95% CI: 0.40-0.72), based on an IFNγ readout after Mtb antigen stimulation. The stratification of the two groups could be further improved by using the Mtb Ag/BCG IFNγ ratio response (p < 0.0001, AUC = 0.918, 95% CI: 0.84-0.98). We also identified additional cytokines that distinguished latent infection from TB disease; and show that the primary differences between the TruCulture and QFT systems were a result of higher levels of non-specific innate immune activation in QFT tubes, due to the lack of a buffering solution in the latter. We conclude that TruCulture offers a next-generation solution for whole blood stimulation and immunomonitoring with the possibility to discriminate active and latently infected persons.