Immunoassays for allergy research
Published: June 23, 2025
5 minute read
Authored by: Jens Gertow
Understanding allergic reactions – from seasonal hay fever to severe food allergies – requires more than just knowing what triggers them. It demands detailed insight into how the immune system responds at the cellular level. That’s where ELISpot steps in.
Allergy needs precision – and ELISpot delivers
Unlike bulk cytokine detection methods, ELISpot allows researchers to identify and enumerate individual cytokine-secreting cells (Akdis et al., 2004; Jutel et al., 2006). This single-cell sensitivity is especially important in allergy research, where low-frequency allergen-specific T and B cells may drive disproportionate clinical symptoms.
The challenge: capturing rare immune responses
Allergic diseases are typically driven by Th2-biased immune responses, characterized by cytokines like IL-4, IL-5, and IL-13 (Romagnani, 2000). But these cytokine-secreting T cells often exist at very low frequencies, especially in peripheral blood. Traditional bulk assays such as ELISA or multiplex bead-based methods often miss these subtle responses, or they struggle to separate background noise from meaningful data.
That’s frustrating when you're working with limited sample volumes from sensitized individuals or pediatric populations. And it can lead to weeks of inconclusive data.
The ELISpot advantage: sensitivity that sees what others miss
With ELISpot, cytokines are captured immediately upon secretion – before they can be degraded, absorbed, or diluted. This means researchers can:
- Detect as few as 1 cell in 100,000, ideal for rare allergen-specific T cells
- Use low sample volumes, preserving precious PBMCs
- Analyze responses to defined allergens (e.g., dust mite peptides, birch pollen, or peanut antigens)
- Evaluate the efficacy of immunotherapies, including allergen-specific desensitization (Scadding et al., 2012)
And because ELISpot offers a quantitative readout (each spot = one secreting cell), it’s easier to compare across cohorts, treatment timepoints, or allergen concentrations.
Nickel-specific IL-4 responses revealed by ELISpot
Peripheral blood mononuclear cells (PBMCs) from individuals with varying degrees of nickel allergy (+ to +++) and from non-allergic controls were stimulated with 50 µM NiCl₂ for 40 hours. IL-4 secretion was evaluated using both ELISpot and ELISA. While ELISA detected responses in only 3 allergic donors, ELISpot identified nickel-reactive IL-4–secreting cells in 23 out of 31 allergic individuals – highlighting its superior sensitivity in detecting allergen-specific T cell responses (in-house data).
Applications across the allergy pipeline
Whether you're working in basic immunology or translational research, ELISpot opens new doors in allergy studies:
- Characterizing Th2 cell responses in asthmatic vs. non-asthmatic donors
- Evaluating allergen immunotherapy responses, especially the emergence of regulatory T cells (Tregs) and IL-10 secretion
- Mapping allergen-specific T cell epitopes to design safer immunotherapeutics
- Monitoring longitudinal immune memory, such as seasonal variation in pollen sensitivity
Tips for success in allergy studies using ELISpot
To get the most out of your allergy-focused ELISpot assays:
- Choose cytokines relevant to Th2-mediated allergy: IL-4, IL-5, IL-13, and IL-10
- Consider dual-color FluoroSpot if you want to distinguish Th2 vs. Treg phenotypes (e.g., IL-5/IL-10). This is particularly relevant when evaluating the potency of indoor allergens where the frequency of reactive T cells can vary greatly between individuals. For example, FluoroSpot can be used to assess cytokine secretion in response to cockroach allergens.
- Use well-characterized peptide pools or whole allergen extracts for stimulation
- Use Mabtech's pre-coated ELISpot Plus kits to reduce variability
- A key hallmark of allergic disease is the involvement of immunoglobulin E (IgE). Upon allergen exposure, IgE binds to FcεRI receptors on mast cells and basophils, initiating degranulation and the release of inflammatory mediators such as histamine. This immediate response contributes to the classic symptoms of allergy. IgE also amplifies adaptive immunity by reinforcing Th2 cell polarization, driving increased secretion of cytokines like IL-4, IL-5, and IL-13. While ELISpot is not ideal to measure IgE directly in this context, Mabtech’s ELISA kits allow for sensitive quantification of total and allergen-specific IgE in serum or plasma. These can be used alongside ELISpot assays to correlate systemic IgE responses with cellular cytokine output – a powerful combination for profiling allergic status or tracking immunotherapy effects.
Mabtech solutions for allergy research
We offer a wide range of validated ELISpot kits, including for IL-4, IL-5, IL-13, IL-10, IL-31, and IFN-γ, in Flex, Plus, and Pro formats. Whether you’re looking to build a custom panel or use pre-coated plates for maximum reproducibility, our kits are designed to simplify your workflow.
And for those ready to scale up? Our ASTOR 2 reader ensures fast, automated spot counting with unmatched accuracy using RAWspot™ technology.
Ready to explore ELISpot for your allergy project?
If you’re navigating the complexities of allergic inflammation and need tools that are as sensitive and reliable as your science demands, ELISpot is the perfect fit. Let us help you find the right kit or reader setup – or even customize an assay for your specific allergen.
References
- Akdis CA, Blesken T, Akdis M, Wüthrich B, Blaser K. Role of interleukin 10 in specific immunotherapy. J Clin Invest. 2004
- Jutel M, Akdis M, Akdis CA. Immunological mechanisms of allergen-specific immunotherapy. Allergy. 2006
- Romagnani S. The role of lymphocytes in allergic disease. J Allergy Clin Immunol. 2000
- Scadding GW, Shamji MH, Jacobson MR, Lee DI, Wilson D, Lima MT, et al. Sublingual grass pollen immunotherapy is associated with increases in sublingual Foxp3-expressing cells and FOXP3 demethylation. J Allergy Clin Immunol. 2012