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Updated: May 14, 2026
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Enzyme-linked Immunospot Assay (ELISPOT): Quantification of Th-1 Cellular Immune Responses Against Microbial Antigens
Published on: November 23, 2010
ELISPOT as a Functional for Biomarker Study in Cancer Immunotherapy: Applications and Future Directions
Laura R Fernández Castro1,2,3, Matias Regiart4, Francisco Gabriel Ortega-Sánchez1,2,3
1GENYO, Centre for Genomics and Oncological Research, Pfizer/University of Granada/Andalusian Regional Government PTS, Avenida de la Ilustración, 114, 18016 Granada, Spain.
International Journal of Molecular Sciences
|May 13, 2026
Summary
The Enzyme-Linked ImmunoSpot (ELISPOT) assay is valuable for monitoring T cell responses in cancer immunotherapy. Standardization is needed for its use as a clinical biomarker.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- The Enzyme-Linked ImmunoSpot (ELISPOT) assay quantifies antigen-specific T cell responses, particularly interferon gamma (IFN-γ) secretion, at a single-cell level.
- It is widely applied in cancer research for evaluating immune responses to various immunotherapies, including vaccines, oncolytic viruses, cellular therapies, and immune checkpoint inhibitors.
Purpose of the Study:
- To systematically review and synthesize evidence on the application of ELISPOT in cancer immunotherapy research.
- To assess the utility of ELISPOT in validating antigens, monitoring T cell activity, and characterizing T cell function in preclinical and clinical settings.
Main Methods:
- A systematic qualitative synthesis of 78 studies was conducted, adhering to PRISMA guidelines.
- Studies were selected from PubMed, Scopus, and Embase, focusing on research with translational relevance in cancer immunotherapy.
Main Results:
- ELISPOT is extensively used to validate tumor antigens, monitor T cell responses during immunotherapy, and characterize T cell function.
- A limited number of studies demonstrate direct links between ELISPOT results and clinical outcomes.
- Significant variability in assay protocols and reporting hinders cross-study comparability and reproducibility.
Conclusions:
- ELISPOT is a valuable tool for immune monitoring in cancer immunotherapy research.
- Further standardization, prospective validation, and integration with other analytical methods are necessary for its clinical biomarker implementation.
