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Related Experiment Video

Updated: Nov 4, 2025

Enzyme-linked Immunospot Assay ELISPOT: Quantification of Th-1 Cellular Immune Responses Against Microbial Antigens
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CTL ELISPOT Assay and T Cell Detection.

Elena Ranieri1, Giuseppe Stefano Netti2, Margherita Gigante2

  • 1Center of Molecular Medicine, Clinical Pathology, Department of Surgical and Medical Sciences, University of Foggia, Foggia, Italy. elena.ranieri@unifg.it.

Methods in Molecular Biology (Clifton, N.J.)
|May 30, 2021
PubMed
Summary

Enzyme-linked immune absorbent spot (Elispot) is a sensitive assay for detecting T-cells. This high-throughput method is crucial for monitoring immune responses in clinical trials and diagnostics.

Keywords:
CancerCytokinesELISPOTT-cell

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Area of Science:

  • Immunology
  • Cellular Biology
  • Biotechnology

Background:

  • Enzyme-linked immune absorbent spot (Elispot) is a quantitative method for measuring T-cell activation parameters.
  • Its high sensitivity allows detection of low-frequency antigen-specific T-cells secreting cytokines and effector molecules.
  • Elispot is widely used in research and diagnostics due to its accuracy, sensitivity, reproducibility, and robustness.

Purpose of the Study:

  • To provide clear protocols for researchers applying the Elispot assay.
  • To detail Interferon-gamma (IFN-γ) and Perforin Elispot assays.
  • To highlight Elispot's utility in monitoring cytotoxic T-cell (CTL) responses and evaluating antigen-specific T-cell immunity.

Main Methods:

  • Elispot assay for quantifying T-cell activation and cytokine secretion.
  • Detection of cytokines such as IFN-γ, IL-2, IL-4, IL-5, IL-10, IL-13, IL-17, and TNF-α.
  • Measurement of effector molecules like granzyme B and perforin.

Main Results:

  • Elispot assays, particularly IFN-γ Elispot, are gold standards for evaluating antigen-specific T-cell immunity in clinical trials.
  • The method enables the distinction of various T-cell subsets (Th1, Th2, Th17) based on cytokine profiles.
  • Elispot demonstrates superior sensitivity compared to other methods for defining antigen-specific lymphocyte frequencies.

Conclusions:

  • Elispot is a reliable and sensitive method for assessing T-cell responses in various diseases including cancer, infections, allergies, and autoimmune diseases.
  • Its high-throughput nature and ability to detect rare antigen-specific T-cells make it invaluable for immune diagnostics and vaccine development.
  • The described protocols for IFN-γ and Perforin Elispot assays offer practical guidance for researchers.