OMIP-035
Functional analysis of natural killer cell subsets in macaques
Cytometry PART A, Volume 89, Issue 9, 799–802 (2016)
Kim L. Weisgrau, Moritz Ries, Nicholas Pomplun, David T. Evans, Eva G. Rakasz
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 | Fluorophore | 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 |
