OMIP-044
28‐color immunophenotyping of the human dendritic cell compartment
Cytometry PART A, Volume 93, Issue 4, 402-405 (2018)
Florian Mair, Martin Prlic
Description: This 28‐color panel has been developed for an extensive phenotyping of antigen‐presenting cells (APCs) in human blood and tissue samples. 15 markers associated with changes in dendritic cell (DC) function were selected based on the current literature. Five additional markers were used to pre-gate on canonical DC subpopulations, namely CD141+ cross‐presenting conventional DC1 (cDC1), CD1c+ conventional DC2 (cDC2), CD141‐ CD1c‐ cDCs as well as CD123+ plasmacytoid DCs (pDC). Furthermore, seven lineage markers were included for parallel enumeration of CD14+ monocytes, B cells, NK cells as well as CD4+ and CD8+ αβ T cells with basic phenotyping of their differentiation status. The panel has been tested on cryopreserved peripheral blood mononuclear cells (PBMCs) and allows further inclusion of DC antigens at the cost of pan‐phenotyping markers.
Species: Human
Tissue: Peripheral blood
Cell Subset: B cell, T cell, Dc, Mdc/cdc, Pdc, Nk, Myeloid cells, Monocyte, Mdc1
Keyword: Cryopreserved pbmc
Machine: BD FACSymphony A5
Panel details
| Marker | Species | Clone | Fluorophore | Purpose | Co-expressions groups | Antigen Density |
|---|---|---|---|---|---|---|
| Dead cells | Human | Live/Dead Fix Blue | Viability | |||
| CD272 | Human | J168-540 | Brilliant Blue 630-P2 | DC phenotype | DC | Med |
| CD8a | Human | RPA-T8 | Brilliant Blue 660-P2 | CD8 T cell | T cell | High |
| CD40 | Human | 5C3 | Brilliant Ultraviolet 395 | DC phenotype | B cell, DC | Med |
| CD56 | Human | NCAM16.2 | Brilliant Ultraviolet 563 | NK | NK | Low |
| CD3 | Human | UCHT1 | Brilliant Ultraviolet 661 | T cell | T cell | High |
| CD86 | Human | FUN-1 | Brilliant Ultraviolet 737 | DC phenotype | B cell, T cell, monocyte, DC | Med |
| CX3CR1 | Human | 2A9-1 | Brilliant Violet 421 | DC phenotype | DC | Low |
| CD85k | Human | ZM3.8 | Brilliant Violet 480 | DC phenotype | mDC | Med |
| CD45RA | Human | HI100 | Brilliant Violet 570 | DC, T cell | T cell | Med |
| CD141 | Human | 1A4 | Brilliant Violet 605 | DC subset | mDC, monocyte | Low |
| CD14 | Human | MfP9 | Brilliant Violet 711 | monocyte | monocyte | High |
| CD11b | Human | ICRF44 | Brilliant Violet 750 | DC phenotype | monocyte, NK, DC, T cell, B cell | Med |
| CD123 | Human | 7G3 | Brilliant Violet 786 | pDC (plasmacytoid DC) | DC | Med |
| CD26 | Human | M-A261 | FITC | DC subset | NK, B cell | Low |
| CD163 | Human | GHI/61 | PE-CF594 | DC phenotype | DC, monocyte | Low |
| CD80 | Human | L307.4 | PE-Cy5 | DC phenotype | B cell, T cell, DC | Low |
| CD19 | Human | SJ25-C1 | PE-Cy5.5 | B cell | B cell | Low |
| CD4 | Human | SK3 | PE-Cy7 | CD4 T cell | T cell | High |
| CD1c | Human | F10/21A3 | Alexa Fluor 647 | DC subset | mDC | Med |
| CD11c | Human | B-ly6 | Alexa Fluor 700 | DC lineage marker | mDC | Med |
| HLA DR | Human | G46-6 | APC-H7 | DC lineage marker | DC, cDC | High |
| CCR7 | Human | G043H7 | PE | DC phenotype, T cell phenotype | T cell | Low |
| CD38 | Human | HIT2 | Brilliant Blue 790-P | DC phenotype | B cell, Monocyte | Low |
| CD45 | Human | HI30 | Brilliant Ultraviolet 805 | hematopoitic cell | monocyte, T cell, B cell, DC, mDC, pDC, NK | High |
| CD172a | Human | SE5A5 | Brilliant Violet 650 | DC phenotype | monocyte, DC | Low |
| CD32 | Human | FLI8.26 | Brilliant Blue 700 | DC phenotype | B cell, monocyte | Med |
| CD16 | Human | 3G8 | Brilliant Ultraviolet 496 | DC phenotype, monocyte phenotype | monocyte | Med |
