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OMIP-008

Measurement of Th1 and Th2 cytokine polyfunctionality of human T cells

Cindy L. Zuleger, Mark R. Albertini Cytometry PART A, Volume 81A, Issue 6, 450-452 (2012)

PURPOSE: This panel was optimized to assess CD4+ and CD8+ T cell responses to various tumor antigens from melanoma patients. The panel was tested on single-cell derived T cell isolates (SCD-T) and T cell lines derived from peripheral blood mononuclear cells (PBMC) from melanoma patients, T cell lines from the tumor environment of melanoma patients, and fresh and cryopreserved PBMC (healthy donors). Staining can be performed in 96-well plates for high-throughput.

CELL TYPE: Peripheral Blood Mononuclear Cells, T cell clones/lines, tumor-resident T cell lines

MACHINE: BD LSR2

Marker Target Clone Fluorochrome Purpose
Dead cells Human Live/Dead Fix Violet viability(dump)
CD8 Human 3B5 Qdot 605 lineage
CD19 Human HIB19 eFluor 450 exclusion
CD3 Human UCHT1 Alexa Fluor 700 lineage
CD4 Human RPA-T4 APC-Cy7 lineage
CD14 Human M5E2 Pacific Blue exclusion
IL-2 Human MQ1-17H12 PerCP-Cy5.5 function/cytokine secretion
IL-4 Human 8D4-8 Alexa Fluor 488 function/cytokine secretion
IL-10 Human JES3-9D7 PE function/cytokine secretion
IFN-gamma Human B27 APC function/cytokine secretion
TNF alpha Human Mab11 PE-Cy7 function/cytokine secretion