Abstract
Rapid and reliable diagnostic work-up of tuberculosis (TB) remains a major healthcare goal. In particular, discrimination of TB infection from TB disease with currently available diagnostic tools is challenging and time-consuming. This study aimed at establishing a standardised blood-based assay that rapidly and reliably discriminates TB infection from disease based on multiparameter analysis of TB antigen-reactive CD4+ T cells acting as sensors for the TB stage-specific immune status.
A total of 157 HIV-negative subjects suspected with TB infection or disease was recruited from local tertiary care hospitals. Peripheral blood mononuclear cells were analysed for CD4+ T cells reactive to the Mycobacterium tuberculosis antigens PPD and ESAT-6/CFP-10. The activation state of TB antigen-reactive T cells, identified by surface expression of CD154, was evaluated according to the expression profile of proliferation marker Ki-67 and activation markers CD38 and HLA-DR. Using data from 81 subjects with clinically confirmed TB infection (n=34) or culture-proven pulmonary or extrapulmonary TB disease (n=47) twelve parameters were derived from the expression profile and integrated into a scoring system.
Using the scoring system our assay allowed reliable discrimination of TB infection from both pulmonary and extrapulmonary TB disease with high sensitivity (90.9%) and specificity (93.3%) as was confirmed by Monte-Carlo cross validation.
With low time requirement, ease of sample collection and high sensitivity and specificity both for pulmonary and extrapulmonary TB disease, we believe this novel standardised TB-Flow Assay will improve the work-up of patients with suspected TB disease supporting rapid TB diagnosis and facilitating treatment decisions.
Abstract
In a prospective study, a scoring system based on analysis of the activation state of tuberculosis (TB)-specific CD4+ T cells was developed that allows reliable discrimination of tuberculosis infection and disease with high sensitivity and specificity.
Footnotes
This manuscript has recently been accepted for publication in the European Respiratory Journal. It is published here in its accepted form prior to copyediting and typesetting by our production team. After these production processes are complete and the authors have approved the resulting proofs, the article will move to the latest issue of the ERJ online. Please open or download the PDF to view this article.
Conflict of interest: Andrej Mantei is listed as inventors in a patent (10 2018 131 696.8) that has been issued and a patent (PCT/EP2019/084392) that is pending on the discrimination of TB infection and tuberculosis disease using the method described herein.
Conflict of interest: Tim Meyer is listed as inventors in a patent (10 2018 131 696.8) that has been issued and a patent (PCT/EP2019/084392) that is pending on the discrimination of TB infection and tuberculosis disease using the method described herein.
Conflict of interest: Mariana Schürmann has nothing to disclose.
Conflict of interest: Christiane Beßler has nothing to disclose.
Conflict of interest: Harald Bias has nothing to disclose.
Conflict of interest: David Krieger has nothing to disclose.
Conflict of interest: Torsten Bauer has nothing to disclose.
Conflict of interest: Petra Bacher is advisor to Miltenyi Biotec who own IP rights on the use of CD154 for antigen-specific T-cell detection. Petra Bacher is listed as inventors in a patent (10 2018 131 696.8) that has been issued and a patent (PCT/EP2019/084392) that is pending on the discrimination of TB infection and tuberculosis disease using the method described herein
Conflict of interest: Johannes Helmuth has nothing to disclose.
Conflict of interest: Hans-Dieter Volk is listed as inventors in a patent (10 2018 131 696.8) that has been issued and a patent (PCT/EP2019/084392) that is pending on the discrimination of TB infection and tuberculosis disease using the method described herein.
Conflict of interest: Dirk Schürmann has nothing to disclose.
Conflict of interest: Alexander Scheffold is advisor to Miltenyi Biotec who own IP rights on the use of CD154 for antigen-specific T-cell detection. Alexander Scheffold is listed as inventors in a patent (10 2018 131 696.8) that has been issued and a patent (PCT/EP2019/084392) that is pending on the discrimination of TB infection and tuberculosis disease using the method described herein.
Conflict of interest: Christian Meisel is listed as inventors in a patent (10 2018 131 696.8) that has been issued and a patent (PCT/EP2019/084392) that is pending on the discrimination of TB infection and tuberculosis disease using the method described herein.
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- Received June 23, 2021.
- Accepted November 26, 2021.
- Copyright ©The authors 2022. For reproduction rights and permissions contact permissions{at}ersnet.org
