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A chemiluminescence assay targeting granzyme A activity for monitoring inflammatory bowel disease

Nature Biomedical Engineering (2026)Cite this article

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Abstract

The diagnosis and monitoring of inflammatory bowel disease (IBD) relies on histologic and endoscopic analysis, as well as measurements of generic markers of inflammation. However, there are no specific tests that report on T cell-mediated immune responses as a key driver of IBD pathogenesis. Here we detect increasing granzyme A (GzmA) in gut biopsies and confirm that CD8+ T cells secrete its active form to induce interleukin (IL)-8. We then rationally design a non-invasive chemiluminescence assay for measuring active GzmA in stool supernatants from patients with IBD. For our assay, we synthesize peptide-based GzmA-specific inhibitors and chemiluminescent reporters and use them to characterize biosamples from ~150 human patients with IBD and healthy controls. Our results demonstrate that GzmA activity is an indicator of gut inflammation that can enhance the identification of patients with IBD over existing tests and potentially act as a mechanistic biomarker for the dominance of T cell activity. We envision that the selectivity and sensitivity of our GzmA activity-based optical assay will accelerate the design of additional biomedical approaches to enhance precision medicine in IBD.

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Fig. 1: GzmA is elevated in the intestinal tract of patients with IBD.
Fig. 2: CD8+ T cells secrete extracellular GzmA to induce the secretion of the neutrophil chemoattractant IL-8.
Fig. 3: Chemical synthesis and characterization of GzmA-INH as a potent and selective inhibitor for hGzmA.
Fig. 4: Synthesis and characterization of GzmA-CL as a hGzmA-activatable reporter.
Fig. 5: An activity-based assay measuring hGzmA in stool supernatants from patients with IBD.

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Data availability

Data from Figs. 1–5, Supplementary Figs. 1–25 and Supplementary Table 1 of this study are available from the corresponding author upon request. Source data are provided with this paper.

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Acknowledgements

We acknowledge funds from an ERC Consolidator Grant (DYNAFLUORS, 771443; M.V.), an ERC PoC grant (IBDIMAGE, 957535; M.V.), an EIC Transition grant (IBDSENSE, 101113123; M. V.) and the Medical Research Council (MR/R01566X/1; D.G.). This project has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement (859908; Z.C.) and from the Leona M. and Harry B. Helmsley Charitable Trust (G-1911-03343; C.S.C., P.L. and G.-T.H.). We thank the Flow Cytometry and Confocal Advanced Light Microscopy facilities at the University of Edinburgh for the technical support. For open access, we applied a CC-BY public copyright license to any author accepted manuscript version arising from this submission. We acknowledge BioRender.com for assistance with figure creation.

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  1. These authors contributed equally: Jamie I. Scott, Zhiming Cheng.

Authors and Affiliations

  1. Centre for Inflammation Research, Institute for Regeneration and Repair, The University of Edinburgh, Edinburgh, UK

    Jamie I. Scott, Zhiming Cheng, Emily J. Thompson, Utsa Karmakar, Verity Cowell, Doireann Gordon, Lorena Mendive-Tapia, Alexander Le Saint-Grant, Cher S. Chuah, Phoebe Lau, Adriano G. Rossi, Gwo-Tzer Ho & Marc Vendrell

  2. IRR Chemistry Hub, Institute for Regeneration and Repair, The University of Edinburgh, Edinburgh, UK

    Jamie I. Scott, Zhiming Cheng, Utsa Karmakar, Doireann Gordon, Lorena Mendive-Tapia, Alexander Le Saint-Grant & Marc Vendrell

  3. Department of Organic Chemistry, School of Chemistry, Faculty of Exact Sciences, Tel Aviv University, Tel Aviv, Israel

    Maya David & Doron Shabat

  4. Department of Gastroenterology and Hepatology, University Medical Centre Groningen, University of Groningen, Groningen, The Netherlands

    Pia Volkmer & Wouter B. Nagengast

  5. School of Infection and Immunity, University of Glasgow, Glasgow, UK

    Gwo-Tzer Ho

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Contributions

J.I.S. performed the biological and spectral validation of GzmA-CL. J.I.S. and V.C. processed the clinical samples and performed the associated experiments with the samples. Z.C., M.D. and L.M.-T. contributed to the synthesis and characterization of materials. D.G. performed the statistical analysis. E.J.T. and U.K. performed the imaging and analysis of tissue biopsies. E.J.T. performed the flow cytometry and cytokine analysis. E.J.T. and U.K. performed the cell culture. D.G., A.L.S.-G., C.S.C. and P.L. performed the sample retrieval and data pooling. P.V. obtained and performed the fresh-frozen biopsy staining. A.G.R., W.B.N., D.S., G.-T.H. and M.V. all contributed to the conceptualization of the study. M.V. wrote the paper with feedback from all authors.

Corresponding author

Correspondence to Marc Vendrell.

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Competing interests

The University of Edinburgh has filed a patent covering methods for granzyme detection, in which J.I.S., D.G. and M.V. are named inventors. M.V. is a current member of the Scientific Advisory Board of Celtarys. The other authors declare no competing interests.

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Scott, J.I., Cheng, Z., Thompson, E.J. et al. A chemiluminescence assay targeting granzyme A activity for monitoring inflammatory bowel disease. Nat. Biomed. Eng (2026). https://doi.org/10.1038/s41551-025-01588-1

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