PT  - JOURNAL ARTICLE
AU  - Felix J. Hartmann
AU  - Joel Babdor
AU  - Pier Federico Gherardini
AU  - El-Ad D. Amir
AU  - Kyle Jones
AU  - Bita Sahaf
AU  - Diana M. Marquez
AU  - Peter Krutzik
AU  - Erika O’Donnell
AU  - Natalia Sigal
AU  - Holden T. Maecker
AU  - Everett Meyer
AU  - Matthew H. Spitzer
AU  - Sean C. Bendall
TI  - Comprehensive Immune Monitoring of Clinical Trials to Advance Human Immunotherapy
AID  - 10.1101/489765
DP  - 2018 Jan 01
TA  - bioRxiv
PG  - 489765
4099  - http://biorxiv.org/content/early/2018/12/13/489765.short
4100  - http://biorxiv.org/content/early/2018/12/13/489765.full
AB  - 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.