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OMIP-035

Functional analysis of natural killer cell subsets in macaques

Kim L. Weisgrau, Moritz Ries, Nicholas Pomplun, David T. Evans, Eva G. Rakasz Cytometry PART A, Volume 89, Issue 9, 799–802 (2016)

PURPOSE: This panel was developed to measure the functional capability of natural killer (NK) cell subsets in rhesus macaques (Macaca mulatta). It includes markers to determine the frequency of cytokine secreting and cytotoxic NK cell subpopulations in peripheral blood mononuclear cell (PBMC) samples stimulated in vitro with human 721.221 cells. NK cell subsets were defined by the expression of killer cell immunoglobulin-like receptors (KIRs) Mamu-KIR3DL01 and Mamu-KIR3DL05, and differentiation antigens CD16 and CD56. The panel can be used to assess the functional capability of NK cells in a range of normal and pathologic conditions of captive bred rhesus macaques of Indian origin.

CELL TYPE: 721.221 stimulated PBMC

MACHINE: SORP BD LSR 2

Marker Target Clone Fluorochrome Purpose
Dead cells Macaca mulatta Live/Dead Fix Near IR (775) viability
CD3 Macaca mulatta SP34-2 PE-CF594 Exclusion
CD8 Macaca mulatta RPA-T8 Brilliant Violet 711 NK subset
CD16 Macaca mulatta 3G8 Pacific Blue NK subset
CD20 Macaca mulatta 2H7 PE-CF594 Exclusion
CD45 Macaca mulatta D058-1283 Brilliant Violet 786 hematopoitic cell lineage
CD56 Macaca mulatta B159 PerCP-Cy5.5 NK subset
CD107a Macaca mulatta H4A3 Brilliant Violet 605 NK cell function
KLRC1/2 Macaca mulatta Z199 PE-Cy7 NK subset
CD158 Macaca mulatta NKVFS1 PE NK subset
KIR3dl05 Macaca mulatta APC NK subset
Granzyme B Macaca mulatta GB11 Brilliant Violet 510 NK cell function
IFN-gamma Macaca mulatta 4S.B3 FITC NK cell function
TNF alpha Macaca mulatta Mab11 Alexa Fluor 700 NK cell function