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OMIP-052

An 18‐Color Panel for Measuring Th1, Th2, Th17, and Tfh Responses in Rhesus Macaques

Mitzi M. Donaldson, Shing‐Fen Kao, Kathryn E. Foulds Cytometry PART A, Volume 95, Issue 3, 261-263 (2019)

PURPOSE: This 18‐color panel was designed to measure vaccine‐induced T cell responses in rhesus macaques by intracellular cytokine staining (ICS). It detects seven cytokines, IFN‐γ, IL‐2, IL‐4, IL‐5, IL‐13, IL‐17, and IL‐21, two memory markers, CD45RA and CCR7, and four follicular helper T cell (Tfh) markers, CXCR3, CXCR5, ICOS, and PD‐1 (Figure 1). While this panel was optimized for use on cryopreserved rhesus macaque peripheral blood mononuclear cells (PBMC) (Table 1), it can also be used on rhesus macaque tissue samples and human PBMC and tissue samples.

CELL TYPE: PBMC

MACHINE: BD LSR2

Marker Target Clone Fluorochrome Purpose
Dead cells Rhesus macaque Live/Dead Fix Aqua Viability
CD3 Rhesus macaque SP34-2 APC-Cy7 T cell
CD4 Rhesus macaque S3.5 PE-Cy5.5 T cell
CD8 Rhesus macaque RPA-T8 Brilliant Violet 570 T cell
CD69 Rhesus macaque TP1.55.3 ECD background reduction
IFN-gamma Rhesus macaque B27 Alexa Fluor 700 cytokine
IL-2 Rhesus macaque MQ1-17H12 Brilliant Violet 750 cytokine
IL-13 Rhesus macaque JES10-5A2 Brilliant Violet 421 cytokine
IL-17 Rhesus macaque BL168 Brilliant Violet 605 cytokine
IL-21 Rhesus macaque 3A3-N2.1 Alexa Fluor 647 cytokine
CD45RA Rhesus macaque 5H9 PE-Cy5 memory marker
ccr7 Rhesus macaque G043H7 Brilliant Violet 650 memory marker
CD183 Rhesus macaque 1C6/CXCR3 Brilliant Violet 711 Tfh
CD185 Rhesus macaque MU5UBEE PE Tfh
PD-1 Rhesus macaque EH12.2H7 Brilliant Violet 785 Tfh
CD278 Rhesus macaque C398.4A PE-Cy7 Tfh
IL-4 Rhesus macaque MP4-25D2 BB700 cytokine
IL-5 Rhesus macaque JES1-39D10 BB515 cytokine