Immune responses to ESAT-6 and CFP-10 by FASCIA and multiplex technology for diagnosis of M. tuberculosis infection; IP-10 is a promising marker

• Published in: PloS one 2012-11, Vol.7 (11), p.e43438-e43438
• Main Authors: Borgström, Emilie, Andersen, Peter, Atterfelt, Fredrik, Julander, Inger, Källenius, Gunilla, Maeurer, Markus, Rosenkrands, Ida, Widfeldt, Maria, Bruchfeld, Judith, Gaines, Hans
• Format: Article
• Language: English
• Subjects: Active control; Adolescent; Adult; Aged; Antigens; Antigens, Bacterial - immunology; Bacterial Proteins - immunology; Biological response modifiers; Biology; Biomarkers; Blood; Chemokine CXCL10 - blood; Chemokine CXCL10 - immunology; Chemokines; Comparative analysis; Cytokines; Cytokines - blood; Cytokines - immunology; Diagnosis; Disease control; ESAT-6 antigen; Fascia; Female; Financial planners; Granulocyte-macrophage colony-stimulating factor; Hospitals; Humans; Immune response; Immunology; Infections; Infectious diseases; Interferon; Interferon gamma; Interleukin 10; Interleukin 12; Interleukin 13; Interleukin 17; Interleukin 2; Interleukin 4; Interleukin 5; Interleukin 6; IP-10 protein; Lung - immunology; Lung - microbiology; Lymphoblasts; Lymphocytes; Male; Medical diagnosis; Medicine; Middle Aged; Multiplexing; Mycobacterium tuberculosis; Mycobacterium tuberculosis - immunology; Mycobacterium tuberculosis - isolation & purification; Patients; Proteins; Skin tests; Stimulation; T cell receptors; Technology; Technology application; Tuberculin; Tuberculosis; Tuberculosis - blood; Tuberculosis - diagnosis; Tuberculosis - immunology; Tuberculosis - microbiology; Tumor necrosis factor-TNF; Tumor necrosis factor-α; Young Adult; γ-Interferon
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0043438

There is a need for reliable markers to diagnose active and latent tuberculosis (TB). The interferon gamma release assays (IGRAs) are compared to the tuberculin skin test (TST) more specific, but cannot discriminate between recent or remote TB infection. Here the Flow-cytometric Assay for Specific Cell-mediated Immune-response in Activated whole blood (FASCIA), which quantifies expanded T-lymphoblasts by flow-cytometric analysis after long-term antigen stimulation of whole blood, is combined with cytokine/chemokine analysis in the supernatant by multiplex technology for diagnosis of Mycobacterium tuberculosis (Mtb) infection. Consecutive patients with suspected TB (n = 85), with microbiologically verified active pulmonary TB (n = 33), extra pulmonary TB (n = 21), clinical TB (n = 11), presumed latent TB infection (LTBI) (n = 23), patients negative for TB (n = 8) and 21 healthy controls were studied. Blood samples were analyzed with FASCIA and multiplex technology to determine and correlate proliferative responses and the value of 14 cytokines for diagnosis of Mtb infection: IFN- γ, IL-2, TNF-α, IP-10, IL-12, IL-6, IL-4, IL-5, IL-13, IL-17, MIP-1β, GM-CSF, IFN-α2 and IL-10. Cytokine levels for IFN-γ, IP-10, MIP-1β, IL-2, TNF-α, IL-6, IL-10, IL-13 and GM-CSF were significantly higher after stimulation with the Mtb specific antigens ESAT-6 and CFP-10 in patients with active TB compared to healthy controls (p