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Application of Long-term cultured Interferon-γ Enzyme-linked Immunospot Assay for Assessing Effector and Memory T Cell Responses in Cattle
Published on: July 11, 2015
Biomarkers of cell-mediated immunity to bovine tuberculosis
Mitchell V Palmer1, Tyler C Thacker1, Meaghan M Rabideau1
1National Animal Disease Center, Agricultural Research Service, United States Department of Agriculture (USDA), Ames, Iowa, USA.
Insights
Researchers evaluated cytokines and chemokines as potential diagnostic biomarkers for bovine tuberculosis (TB) caused by Mycobacterium bovis (M. bovis) infection in cattle. CXCL9, CXCL10, IL-21, and IL-13 show promise for TB diagnosis in cattle.
Area of Science:
- Veterinary immunology
- Infectious disease diagnostics
- Bovine tuberculosis research
Background:
- Interferon gamma (IFN-γ) release assays (IGRAs) are used for diagnosing tuberculosis (TB) in cattle and humans.
- Current diagnostic methods for bovine TB (bTB) require improvement for early and accurate detection.
- Mycobacterium bovis (M. bovis) is the causative agent of TB in cattle, posing a significant zoonotic risk.
Purpose of the Study:
- To identify novel diagnostic biomarkers for Mycobacterium bovis infection in cattle.
- To evaluate a panel of cytokines and chemokines as potential readouts for bovine TB.
- To assess the kinetic response to M. bovis antigens using gene expression and protein assays.
Main Methods:
- Aerosol infection model of cattle with M. bovis.
- Gene expression profiling to analyze cytokine and chemokine kinetics.
- Purified protein derivative (PPD) and recombinant M. bovis antigens (ESAT-6, CFP10, Rv3615c) used for stimulation.
- Enzyme-linked immunosorbent assay (ELISA) to measure protein levels of selected biomarkers.
Main Results:
- Multiple cytokines and chemokines, including IFN-γ, CXCL9, CXCL10, CCL2, CCL3, TNF-α, IL-1α, IL-1β, IL-1Ra, IL-22, IL-21, and IL-13, were measured.
- Gene expression analysis revealed dynamic responses to M. bovis antigens.
- Protein levels of IFN-γ, CXCL9, and CXCL10 were quantified.
- CXCL9, CXCL10, IL-21, and IL-13 demonstrated potential as diagnostic markers.
Conclusions:
- CXCL9 and CXCL10 show strong potential as diagnostic biomarkers for M. bovis infection in cattle.
- IL-21 and IL-13 also warrant further investigation for bovine TB diagnostics.
- The study highlights the utility of a multi-biomarker approach for improved cattle TB detection.
Abstract:
Whole blood based assays, particularly interferon gamma (IFN-γ) release assays (IGRAs), are used for the diagnosis of both bovine and human tuberculosis (TB). The aim of the current study was to evaluate a panel of cytokines and chemokines for potential use as diagnostic readouts indicative of Mycobacterium bovis (M. bovis) infection in cattle. A gene expression assay was used to determine the kinetics of the response to M. bovis purified protein derivative and a fusion protein consisting of ESAT-6, CFP10, and Rv3615c upon aerosol infection with ∼104 cfu of M. bovis. The panel of biomarkers included: IFN-γ, CXCL9, CXCL10, CCL2, CCL3, TNF-α, IL-1α, IL-1β, IL-1Ra, IL-22, IL-21 and IL-13. Protein levels of IFN-γ, CXCL9, and CXCL10 were determined by ELISA. Findings suggest that CXCL9, CXCL10, IL-21, IL-13, and several acute phase cytokines may be worth pursuing as diagnostic biomarkers of M. bovis infection in cattle.
