Potential of the dual IFN-γ/IL-2 fluorescence-immunospot assay to distinguish different stages in bovine tuberculosis

Sabine Steinbach, H. Martin Vordermeier, Gareth J. Jones*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

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Human studies have identified the potential of measuring Mycobacterium tuberculosis specific IFN-γ and/or IL-2 secreting T cell subsets to distinguish different clinical stages of human tuberculosis (TB). To assess these functional T cell subsets in different states of bovine TB we have established a bovine dual IFN-γ/IL-2 fluorescence-immunospot (FluoroSpot) assay and analysed the frequencies of Mycobacterium bovis (M. bovis) specific IL-2 and/or IFN-γ producing cells in PBMC from 30 cattle naturally infected with M. bovis. Depending on their post mortem results the animals were grouped in 22 cattle with visible lesions (VL) and 8 cattle without visible lesions (NVL). In response to bovine tuberculin purified protein derivative (PPD-B) the frequencies of cytokine producing cells and proportions of IL-2 single producers were significantly higher in VL compared to NVL while PWM-induced cytokine responses were similar between the two groups. Dual IL-2+IFN-γ+ T cells could be identified as the largest PPD-B responsive T cell subset in both cattle groups. In conclusion, our FluoroSpot is a valid method to enumerate individual antigen-specific IFN-γ+ and IL-2+ T cell subsets ex vivo. The greater levels of single IL-2 producing T cells associated with the presence of pathology could be a potential biomarker for active TB in cattle.

Original languageEnglish
Article number109930
Number of pages6
JournalVeterinary Immunology and Immunopathology
Early online date13 Aug 2019
Publication statusPublished - 01 Nov 2019


  • Bovine tuberculosis
  • FluoroSpot
  • IFN-γ
  • IL-2
  • Mycobacterium bovis/immunology
  • Color
  • Enzyme-Linked Immunospot Assay/veterinary
  • Fluorescence
  • Interleukin-2/immunology
  • Interferon-gamma/immunology
  • Tuberculosis, Bovine/diagnosis
  • Animals
  • T-Lymphocyte Subsets/immunology
  • Cattle
  • Antigens, Bacterial/immunology


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