Novel methods for the design, analysis and implementation of ELISpot experiments

The enzyme-linked immunospot (ELIspot) assay is used to quantify the number of cytokine secreting cells and is the focus of numerous basic, clinical and translational studies. The results from an ELIspot assay can be used as an outcome in early stage trials for comparing the immunogenicity of different therapies or as a short-term indicator of long-term clinical benefit of an immunotherapy. Despite its widespread use, there is little consistency in how ELIspot data are analyzed. Numerous proposals have been made that define immunologic response as a dichotomous endpoint (response/no response) based on empirical rules for classifying response as a combination of ELIspot count and negative control values. The multiplicity of definitions makes it difficult, if not impossible, to compare results across studies. Moreover, the use of the ELIspot results as a continuous endpoint should have greater statistical power than a dichotomous endpoint and may allow researchers greater opportunity to discover how patterns over time are associated with long-term outcome. This project proposes a method for analyzing ELIspot data as a continuous measure that adjusts ELIspot count for negative and positive controls. We have data from numerous clinical studies that we can use as a basis for comparing the statistical power of our method with existing methods. Furthermore, we will assess whether patterns of immunologic response are associated with long-term clinical benefit. In the process of improving the analysis of data from ELIspot assays, we will propose several innovative statistical methods that can be used in a wide range of applications. Project Number: 1R01CA299473-01A1 | Fiscal Year: 2026 | NIH Institute/Center: National Cancer Institute (NCI) | Principal Investigator: Mark Conaway (+1 co-PI) | Institution: UNIVERSITY OF VIRGINIA, CHARLOTTESVILLE, VA | Award Amount: $484,387 | Activity Code: R01 | Study Section: Analytics and Statistics for Population Research Panel A Study Section[ASPA] View on NIH RePORTER: https://reporter.nih.gov/project-details/11294031