Th163 - Successful Measurement of 13 Functional Readouts Including IL-5, IL-10, and IL-13 from Human Peripheral Blood Mononuclear Cells via Use a Cytof Intracellular Cytokine Staining Panel

Erika Smith – Virology, Immunology, and Microbiology – Boston University Chobanian & Avedisian School of Medicine; Michael Cohen – Standard BioTools Canada; Wenxi Xu – Standard BioTools Canada; Jiabao Zhong – Virology, Immunology, and Microbiology – Boston University Chobanian & Avedisian School of Medicine; Thirumahal Selvanantham – Standard BioTools Canada; David King – Standard BioTools Canada; Christina Loh – Standard BioTools Canada; Anna Belkina – Pathology and Laboratory Medicine and Flow Cytometry Core Facility – Boston University Chobanian & Avedisian School of Medicine; Jennifer Snyder-Cappione – Virology, Immunology, and Microbiology – Boston University Chobanian & Avedisian School of Medicine

Abstract Text: Measurement of functional signatures of immune cells in a comprehensive, cross-lineage manner would provide key insights into several facets of immunology. Intracellular cytokine staining (ICS) is ideal for this type of analysis, and with the advent of full spectrum flow cytometry and improvements in cell interrogation efficiency with CyTOF, we posited that one or both platforms are now able to measure a large number of functions from individual cells with new resolution. We evaluated stimulation-induced cytokine signal from peripheral blood mononuclear cells in large (31-color) ICS Aurora and CyTOF XT panels that include analytes known to be challenging for cytometric analyses, such as IL-5, IL-10, IL-13, IL-17A, IL-21, IL-22 and GM-CSF. Data collected from the both instruments showed well-resolved signal for many cytokines, including IL-4, IFN-γ, TNF-α, IL-17A, and Granzyme B. However, IL-5, IL-10, IL-13, and GM-CSF positive populations were clearly identified by the CyTOF XT in a stimulation-specific manner but could not be resolved by spectral flow cytometry (in these and/or previous experiments). The CyTOF results can serve as a gold standard to further functional readout capacity using spectral flow ICS panels; however, it is possible that inherent features of fluorescent cytometry may be prohibitive for achievement of equivalent results. Also, with the CyTOF’s capability for further panel expansion (50+ parameters), additional functional readouts could be added to this panel. In sum, our data indicate that the CyTOF XT platform could serve as a catalyst for seminal discoveries germane to the cross-lineage functional diversity of many immune cell subsets.