Abstract
Background: IFN-γ and IL-2 cytokine-profiles define three functional T-cell subsets which may correlate with pathogen load in chronic intracellular infections. We therefore investigated the feasibility of the immunospot platform to rapidly enumerate T-cell subsets by single-cell IFN-γ/IL-2 cytokine-profiling and establish whether immunospot-based T-cell signatures distinguish different clinical stages of human tuberculosis infection.
Methods: We used fluorophore-labelled anti-IFN-γ and anti-IL-2 antibodies with digital overlay of spatially-mapped colour-filtered images to enumerate dual and single cytokine-secreting M. tuberculosis antigen-specific T-cells in tuberculosis patients and in latent tuberculosis infection (LTBI). We validated results against established measures of cytokine-secreting T-cells.
Results: Fluorescence-immunospot correlated closely with single-cytokine enzyme-linked-immunospot for IFN-γ-secreting T-cells and IL-2-secreting T-cells and flow-cytometry-based detection of dual IFN-γ/IL-2-secreting T-cells. The untreated tuberculosis signature was dominated by IFN-γ-only-secreting T-cells which shifted consistently in longitudinally-followed patients during treatment to a signature dominated by dual IFN-γ/IL-2-secreting T-cells in treated patients. The LTBI signature differed from active tuberculosis, with higher proportions of IL-2-only and IFN-γ/IL-2-secreting T-cells and lower proportions of IFN-γ-only-secreting T-cells.
Conclusions: Fluorescence-immunospot is a quantitative, accurate measure of functional T-cell subsets; identification of cytokine-signatures of pathogen burden, distinct clinical stages of M. tuberculosis infection and long-term immune containment suggests application for treatment monitoring and vaccine evaluation.
Original language | English |
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Article number | e15619 |
Pages (from-to) | 1-11 |
Number of pages | 11 |
Journal | PLoS ONE |
Volume | 5 |
Issue number | 12 |
DOIs | |
Publication status | Published - 14 Dec 2010 |
Keywords
- Adolescent
- Adult
- Aged
- Case-Control Studies
- Cell Count
- Cross-Sectional Studies
- Female
- Flow Cytometry/methods
- Humans
- Interferon-gamma/metabolism
- Interleukin-2/metabolism
- Male
- Microscopy, Fluorescence/methods
- Middle Aged
- Mycobacterium tuberculosis/metabolism
- Risk Factors
- T-Lymphocytes/cytology
- Tuberculosis/blood