Enumeration of Functional T-Cell Subsets by Fluorescence-Immunospot Defines Signatures of Pathogen Burden in Tuberculosis
| Autor: | Samuel Bremang, Onn Min Kon, Ajit Lalvani, Kerry A. Millington, D W Connell, Deena Blumenkrantz, Damien J. Montamat-Sicotte, Rosalyn Casey, Murphy Magtoto, Saranya Sridhar, Melissa Wickremasinghe |
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| Jazyk: | angličtina |
| Rok vydání: | 2010 |
| Předmět: |
Bacterial Diseases
Male Vaccine evaluation T-Lymphocytes Immunofluorescence lcsh:Medicine Cell Count Adaptive Immunity Cohort Studies 0302 clinical medicine Risk Factors Longitudinal Studies lcsh:Science Pathogen Immune Response 0303 health sciences Multidisciplinary biology Latent tuberculosis T Cells Vaccination Middle Aged Flow Cytometry 3. Good health medicine.anatomical_structure Infectious Diseases Observational Studies Medicine Cytokines Female Antibody Research Article Adult Tuberculosis Adolescent Clinical Research Design T cell Immune Cells Immunology Mycobacterium Mycobacterium tuberculosis Immune Activation 03 medical and health sciences Interferon-gamma Immune system Vaccine Development medicine Humans Biology Immunity to Infections 030304 developmental biology Aged lcsh:R Immunity Immune Defense medicine.disease biology.organism_classification Virology Cross-Sectional Studies Microscopy Fluorescence Immune System Case-Control Studies biology.protein Immunologic Techniques Interleukin-2 lcsh:Q Clinical Immunology 030215 immunology |
| Zdroj: | PLoS ONE PLoS ONE, Vol 5, Iss 12, p e15619 (2010) |
| ISSN: | 1932-6203 |
| Popis: | Background: IFN-c 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-c/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-c and anti-IL-2 antibodies with digital overlay of spatially-mapped colourfiltered 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-c-secreting T-cells and IL-2-secreting T-cells and flow-cytometry-based detection of dual IFN-c/IL-2-secreting T-cells. The untreated tuberculosis signature was dominated by IFN-c-only-secreting T-cells which shifted consistently in longitudinally-followed patients during treatment to a signature dominated by dual IFN-c/IL-2-secreting T-cells in treated patients. The LTBI signature differed from active tuberculosis, with higher proportions of IL-2-only and IFN-c/IL-2-secreting T-cells and lower proportions of IFN-c-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. |
| Databáze: | OpenAIRE |
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