Immune Profiling Enables Stratification of Patients With Active Tuberculosis Disease or Mycobacteriu m tuberculosis Infection

Abstract

Background Tuberculosis (TB) is caused by Mycobacterium tuberculosis (Mtb) infection and is a major public health problem. Clinical challenges include the lack of a blood-based test for active disease. Current blood-based tests, such as QuantiFERON (QFT) do not distinguish active TB disease from asymptomatic Mtb infection. Methods We hypothesized that TruCulture, an immunomonitoring method for whole blood stimulation, could discriminate active disease from latent Mtb infection (LTBI). We stimulated whole blood from active TB patients and compared to LTBI donors. Mtb- specific antigens and live bacillus Calmette-Guerin (BCG) were used as stimuli, with direct comparison to QFT. Protein analyses were performed using conventional and digital ELISA, as well as Luminex. Results TruCulture showed discrimination of active TB cases from LTBI (p < 0.0001 AUC = 0.81) as compared to QFT (p = 0.45 AUC = 0.56), based on an IFNγ readout after Mtb antigen stimulation. This result was replicated in an independent cohort (AUC = 0.89). In exploratory analyses, TB stratification could be further improved by the Mtb Ag/BCG IFNγ ratio (p < 0.0001 AUC = 0.91). Finally, the combination of digital ELISA and transcriptional analysis showed that LTBI donors with high IFNγ clustered with TB patients, suggesting the possibility to identify sub-clinical disease. Conclusions TruCulture offers a next-generation solution for whole blood stimulation and immunomonitoring with the possibility to discriminate active and latent infection.

Bovine innate immune phenotyping via a standardized whole blood stimulation assay

“…Yet our knowledge of the immune response in livestock species lags significantly behind what we know about other traits of agricultural importance including fertility, milk yield and growth traits. The advent of new technologies has enabled a more detailed characterization of the architecture of the immune response in humans enabling identification of susceptibility and disease-associated immune response profiles 12 . Specifically, in terms of profiling cytokine responses, commercial whole blood stimulation systems facilitate longitudinal profiling of responses in the absence of stimulation as well as in response to disease-relevant PAMPs 8 .…”

“…The individuals used in this study are also part of a larger cohort in which detailed health traits have been recorded for each individual in order to assess the risk factors involved in disease susceptibility and to identify the key contributors to human immunological variance 10 . Already, cytokine signatures found using these assays have been linked with disease—including diabetes 11 , and enabled the discrimination between clinical and latent TB 12 . Impaired cytokine responses to infection also leads to increased disease severity, most recently shown in COVID-19 patients 13 , showing additional disease relevance.…”

Whole‐blood cytokine secretion assay as a high‐throughput alternative for assessing the cell‐mediated immunity profile after two doses of an adjuvanted SARS‐CoV‐2 recombinant protein vaccine candidate

“…Alternatively, whole blood can be used to assess CMI in a cytokine secretion assay that is initiated in a simple procedure at the clinical site or local laboratory and then shipped cryopreserved to a central location 19 . Such stimulated whole‐blood cytokine secretion assays are similar to the IFN‐γ release assays used to detect cellular responses to tuberculosis infection, and may include multiplexed measurement of multiple cytokines 20‐22 …”

Tuberculosis alters immune-metabolic pathways resulting in perturbed IL-1 responses

“…We and others have reported distinct secretion of IL-1 agonists and antagonists upon M.tb infection ( 10 , 20 , 62 ), however, whether IL-1 signalling itself is impaired in TB disease remains poorly explored. Here, we report higher expression of most IL-1β-induced genes in TB compared to LTBI before treatment ( Figure 6A , left panel).…”