In-Depth Immunophenotyping With Mass Cytometry During TB Treatment Reveals New T-Cell Subsets Associated With Culture Conversion

Abstract

Tuberculosis (TB) is a difficult-to-treat infection because of multidrug regimen requirements based on drug susceptibility profiles and treatment observance issues. TB cure is defined by mycobacterial sterilization, technically complex to systematically assess. We hypothesized that microbiological outcome was associated with stage-specific immune changes in peripheral whole blood during TB treatment. The T-cell phenotypes of treated TB patients were prospectively characterized in a blinded fashion using mass cytometry after Mycobacterium tuberculosis (Mtb) antigen stimulation with QuantiFERON-TB Gold Plus, and then correlated to sputum culture status. At two months of treatment, cytotoxic and terminally differentiated CD8+ T-cells were under-represented and naïve CD4+ T-cells were over-represented in positive- versus negative-sputum culture patients, regardless of Mtb drug susceptibility. At treatment completion, a T-cell immune shift towards differentiated subpopulations was associated with TB cure. Overall, we identified specific T-cell profiles associated with slow sputum converters, which brings new insights in TB prognostic biomarker research designed for clinical application.

Opti-4TB: A protocol for a prospective cohort study evaluating the performance of new biomarkers for active tuberculosis outcome prediction

“…In addition, 100 μL of cell suspension will be aliquoted for unstained controls. A previously described 29-marker panel of antibodies with the addition of a CD19 marker will be used for staining ( 18 ). Briefly, the panel contains 28 T-cell oriented surface markers (lineage markers, chemokine receptors, activation markers and exhaustion markers), one intracellular target (perforin) and CD19 to improve resolution (antibodies purchased from BD Bioscience, USA, BioLegend, USA and ThermoFisher Scientific, USA).…”

“…Recently, based on an in-depth phenotyping with mass cytometry of peripheral Mtb-specific T-cells, our group found a differential profile according to sputum conversion. We observed an under-representation of terminal memory effector CD8+ T-cell subpopulations (T EMRA ) in patients whose sputum cultures were still Mtb-positive at two months of treatment compared to patients whose cultures were Mtbnegative (18). Successful mycobacterial clearance is linked to CD8+ T-cell effectors, which in turn require CD4+ T-cell engagement (19).…”

New Insights into Biomarkers for Evaluating Therapy Efficacy in Pulmonary Tuberculosis: A Narrative Review

“…The differentiation of T cell subsets upon completion of PTB treatment has been associated with a cure [ 102 ]. A study on PBMCs from patients with PTB undergoing ATTB treatment found that IFNγ + CD4 + T cells (stimulated with CFP10/ESAT-6) increased after 2 weeks of treatment, while IFNγ + CD8 + T cells (stimulated with CFP10/ESAT-6) [ 103 ] and total TNFα + CD8 + T cells (stimulated with Ag85) decreased.…”

Exploratory mass cytometry analysis reveals immunophenotypes of cancer treatment-related pneumonitis

“…This technique provides a unique advantage over traditional flow cytometry, which is limited by spectral overlap and the number of parameters that can be analyzed simultaneously. Mass cytometry has been used to study a variety of biological systems, including immune-mediated diseases, such as cancer and autoimmune conditions (Matsubara et al, 2021) (Hata et al, 2023) (Couloume et al, 2021) (Chedid et al, 2022). Through high-throughput analysis of large numbers of single cells, mass cytometry can provide a more comprehensive understanding of the cellular heterogeneity and signaling pathways involved in these diseases, allowing for the identification of potential therapeutic targets and biomarkers.…”