Equine ELISPOT assay to study secretion of IFNγ and IL-4 from peripheral blood mononuclear cells.

This research aims to expand the use of ELISPOT assays, traditionally used on human and mouse immune cells, to include horse (equine) cells. The study investigates the variations of IFNγ and IL-4 secretions among different horses, potentially serving as a valuable health and diagnostic marker.

Introduction

• The study focuses on expanding the use of the Enzyme Linked ImmunoSpot (ELISPOT) assay beyond its current primary use with human and mouse immune system cells.
• The ELISPOT assay is a tool instrumental in the detection of protein secretion at single cell level, making it crucial for immunological research.

Method

• The researchers developed matched antibody pairs which were then put to use in kits designed for studying the frequency of equine IFNγ- and IL-4-secreting peripheral blood mononuclear cells (PBMCs).
• The equine PBMCs were stimulated with either Concanavalin A (Con A) or more complex substances: calcium ionomycin mixed with phorbol 12-myristate 13-acetate (CaI + PMA).

Results

• The team found that Con A in general had a stronger stimulating effect on the secretion of IL-4 than CaI + PMA.
• Regarding IFNγ secretion, both stimulatory and inhibitory effects were observed.
• The data collected showed a wide dynamic range in IFNγ and IL-4 secretion among different horse donors.

Conclusion

• The results suggest a potential for the ELISPOT assay to be expanded for use with equine cells.
• Furthermore, the broad variation in IFNγ and IL-4 secretion across different horses could serve as a significant health and diagnostic marker, aiding in the detection and treatment of equine diseases.