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Comprehensive Immune Monitoring of Clinical Trials to Advance Human Immunotherapy

By Felix J Hartmann, Joel Babdor, Pier Federico Gherardini, El-Ad D Amir, Kyle Jones, Bita Sahaf, Diana M Marquez, Peter Krutzik, Erika O'Donnell, Natalia Sigal, Holden T Maecker, Everett Meyer, Matthew H. Spitzer, Sean C Bendall

Posted 13 Dec 2018
bioRxiv DOI: 10.1101/489765

The success of immunotherapy has led to a myriad of new clinical trials. Connected to these trials are efforts to discover biomarkers providing mechanistic insight and predictive signatures for personalization. Still, the plethora of immune monitoring technologies can face investigator bias, missing unanticipated cellular responses in limited clinical material. We here present a mass cytometry workflow for standardized, systems-level biomarker discovery in immunotherapy trials. To broadly enumerate human immune cell identity and activity, we established and extensively assessed a reference panel of 33 antibodies to cover major cell subsets, simultaneously quantifying activation and immune checkpoint molecules in a single assay. The resulting assay enumerated ≥ 98% of peripheral immune cells with ≥ 4 positively identifying antigens. Robustness and reproducibility were demonstrated on multiple samples types, across research centers and by orthogonal measurements. Using automated analysis, we monitored complex immune dynamics, identifying signatures in bone-marrow transplantation associated graft-versus-host disease. This validated and available workflow ensures comprehensive immunophenotypic analysis, data comparability and will accelerate biomarker discovery in immunomodulatory therapeutics.

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